CN106074613A

CN106074613A - The application of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function

Info

Publication number: CN106074613A
Application number: CN201610397313.4A
Authority: CN
Inventors: 赵越
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-06-06
Filing date: 2016-06-06
Publication date: 2016-11-09

Abstract

The present invention relates to the application of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function, described photosynthetic bacteria is the hydrogenlike silicon ion (Rhodobacter sphaeroides) that the red antibacterial of Purple Non-sulfur flora belongs to.More specifically photosynthetic bacteria application in the Mylabris conversional solution with anti-lung cancer, hepatocarcinoma, uterus carcinoma effect.Photosynthetic bacteria is applied the bioconversion being used in Mylabris extracting solution Mylabris extracting solution by the present invention first, research proves that photosynthetic bacteria has the effect of attenuation synergistic to Mylabris extracting solution, the toxicity of the Mylabris conversional solution obtained after bioconversion reduces, anticancer effect strengthens, gained Mylabris conversional solution next step can attempt the prevention for people's hepatocarcinoma, pulmonary carcinoma and uterus carcinoma and treatment.

Description

The application of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function

Technical field

The present invention relates to microorganism field, be specifically related to answering of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function With.

Background technology

Mylabris is the Meloidae insecticide Mylabris phalarata Mylabris black mylabris cichorii of phalerata Pallas or yellow The dry body of Mylabris cichorii Linnaeus, its property is hot very toxic, returns liver, stomach, kidney channel.For oral administration have counteracting toxic substances, by Stasis and resolving masses, antineoplastic action.The main component cantharidin of Mylabris and derivant thereof are to kinds cancer especially primary hepatocarcinoma There is the therapeutic effect of affirmative.It has stronger toxicity, reacts based on digestive system and urinary system toxicity, serious appearance Acute renal failure and toxic shock, and toxicity with dosage increase and strengthen, due to Mylabris acute toxicity limit It is in clinical application.How practical clinical uses with forms such as compound recipe, cantharidin, cantharidin derivatives.And from Since Gu, the research for Mylabris attenuation all concentrates on the toxicity being reduced Mylabris by traditional concocting, and makes the cantharidin liter of part China and content reduce thus reduce the toxicity of Mylabris.It addition, also minority scholar proposes to process speckle with the NaOH aqueous alkali of low concentration Cantharis, the cantharidate sodium lower to cantharidin being changed into toxicity.

Method of the prior art the most all cannot meet and keeps while the effective toxicity removing Mylabris or improve The anti-cancer properties of Mylabris；Therefore, still need to study a kind of effective Mylabris toxicity that reduces and improve method and the product of its anti-cancer properties simultaneously Product.

Summary of the invention

It is an object of the invention to overcome the deficiencies in the prior art, it is provided that photosynthetic bacteria attenuation in Mylabris anticancer function The application of potentiation, photosynthetic bacteria has the effect of attenuation synergistic to Mylabris extracting solution, the Mylabris conversional solution obtained after bioconversion Toxicity reduce, anticancer effect strengthens, gained Mylabris conversional solution next step can be used for the prevention of people's hepatocarcinoma, pulmonary carcinoma and uterus carcinoma And treatment.

Another object of the present invention is to provide the preparation method of a kind of Mylabris conversional solution with anticancer function.

Another object of the present invention is to the Mylabris conversional solution providing above-mentioned preparation method to prepare.

Another object of the present invention is to provide a kind of Mylabris biological preparation with anticancer function.

For achieving the above object, the present invention adopts the following technical scheme that

The application of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function, described photosynthetic bacteria is that Purple Non-sulfur flora is red The hydrogenlike silicon ion (Rhodobacter sphaeroides) that antibacterial belongs to.

Preferably, photosynthetic bacteria application in the Mylabris conversional solution with anti-lung cancer, hepatocarcinoma, uterus carcinoma effect.

Preferably, photosynthetic bacteria answering in hepatoma cell strain HepG-2, BEL-7402, SMCC-7211, QGY-7701 With；Photosynthetic bacteria application in lung cancer cell types；Photosynthetic bacteria application in uterine cancer cells strain Hela.

Photosynthetic bacteria in the present invention is purchased from Institute of Microorganism, Academia Sinica, its preservation entitled Purple Non-sulfur flora The hydrogenlike silicon ion (Rhodobacter sphaeroides) that red antibacterial belongs to, deposit number is CGMCC 1.2174.

A kind of preparation method of the Mylabris conversional solution with anticancer function, described preparation method specifically includes following steps:

S1: extract powder of cantharide with extracting solution and obtain Mylabris extracting solution；

S2: concentrated extracting solution also inoculates above-mentioned photosynthetic bacteria, is subsequently adding PSB culture medium and carries out bioconversion；

S3: bioconversion removes photosynthetic bacterial thallus after terminating and i.e. obtains Mylabris conversional solution.

Preferably, described photosynthetic bacteria is the photosynthetic bacteria being in logarithmic (log) phase, the inoculum concentration of described photosynthetic bacteria be 7～ 10%.

Preferably, the time of the bioconversion in described step S2 is 8～12 days.

Preferably, the mass concentration of Mylabris extracting solution described in step S1 is 7～10mg/mL.

Preferably, the extracting solution in step S1 is water；In step S2, the volume of concentrated extracting solution is to the 5 of conversional solution cumulative volume ～8%；Preferably, the pH of described conversional solution system is 6～9.

The Mylabris conversional solution that said method prepares.

A kind of Mylabris biological preparation with anticancer function, described Mylabris biological preparation contains the speckle described in claim 9 Cantharis conversional solution.

Compared with prior art, there is advantages that

Photosynthetic bacteria is applied the bioconversion being used in Mylabris extracting solution Mylabris extracting solution by the present invention first, research Prove that photosynthetic bacteria has the effect of attenuation synergistic to Mylabris extracting solution, the toxicity fall of the Mylabris conversional solution obtained after bioconversion Low, anticancer effect strengthens, gained Mylabris conversional solution next step prevention that can be used for people's hepatocarcinoma, pulmonary carcinoma and uterus carcinoma and treatment.

Accompanying drawing explanation

Fig. 1 is the OD of photosynthetic bacteria in different conversional solution_n/OD₀Growth change curve；

Fig. 2 is the growth curve of photosynthetic bacteria；

Fig. 3 is the growing state of photosynthetic bacteria under different Mylabris extracting solution mass concentration；

Fig. 4 is Mylabris Aqueous extracts group mice accumulation death toll time graph；

Fig. 5 is Mylabris conversional solution group mice accumulation death toll time graph；

Fig. 6 is that Mylabris conversional solution is to the dose effect curve after HepG-2 cell line 24h, 48h, 72h；

Fig. 7 is that Mylabris conversional solution is to the dose effect curve after hepatocarcinoma hela cell 24h, 48h, 72h；

Fig. 8 is that Mylabris conversional solution is to the dose effect curve after lung cancer A549 cell 24h, 48h, 72h；

Fig. 9 is that Mylabris conversional solution is to the dose effect curve after hepatocarcinoma QGY-7701 cell 24h, 48h, 72h；

Figure 10 is that Mylabris conversional solution is to the dose effect curve after hepatocarcinoma SMC-7211 cell 24h, 48h, 72h.

Detailed description of the invention

Below in conjunction with specific embodiment, the present invention is further explained, but specific embodiment is not to the present invention It is limited in any way.Unless stated otherwise, all present invention provide embodiment in, it is provided that raw material all can from market buying obtain ?.

The preparation of embodiment 1 Mylabris extracting solution

(1) medicine and reagent

Mylabris is the dry polypide of Meloidae insecticide Mylabris phalarata Mylabris phalerata Pallas, is purchased from peace Fructus Liquidambaris prepared slices of Chinese crude drugs company limited of state；Hyclone (Hangzhou Ilex purpurea Hassk.[I.chinensis Sims product)；(Hyclone produces RPMI1640 culture fluid Product)；0.25%Trypsin-EDTA (U.S.'s GIBCO product)；CCK-8 test kit (green skies Products)；PSB (the U.S. GIBCO product)；Ethanol (AR).

(2) cell strain

Human liver cancer cell HepG-2, human liver cancer cell BEL-7402, human lung cancer cell A549 are by Guangdong Pharmaceutical University's life Science provides with life pharmacy institute.

1. key instrument

GalaxyS+ CO2 gas incubator (Hua Yuehang Instrument Ltd.)；Chongqing Austria spy's inverted biological microscope BDS200；BIO-RAD Model-1680 microplate reader；Liquid-transfering gun (Eppendorf company)；0.22 μm filter (U.S. Pall).

(4) preparation of each conversional solution of Mylabris

Accurately weigh equal in quality Mylabris coarse powder (will Mylabris pulverize after cross 30 mesh sieves, 50 DEG C be dried 6 hours, obtain Mylabris Coarse powder) 4 parts, add the water of 20 times amount, 55% ethanol, 75% ethanol, 95% ethanol solution respectively, soak 30min, be heated to reflux Extract 2h, cool down, filter, obtain each extracting solution successively.

Each extracting solution is evaporated to 5%～8% less than conversional solution cumulative volume, inoculates 7%～10% (volume ratio) Being in the photosynthetic bacteria (PSB) of logarithmic (log) phase, addition PSB culture medium is the most volume required, and pH=6～9 obtains accordingly with Mylabris medical material Calculating concentration is, conversional solution I, II, III, IV of 7～10mg/mL.Being added without the PSB normal growth group of Mylabris extracting solution it is Matched group, is placed in growth cabinet, Anaerobic culturel, bioconversion 8～12 days.

Timing every day measures the optical density OD value of PSB in each conversional solution and matched group at 660nm, with PSB in conversional solution Cell concentration change compare OD_n/OD₀(OD₀Initial optical density, OD_nThe metabolism optical density of n days) reflect photosynthetic bacterium growth feelings Condition, draws the growth change curve of PSB.

At the end of bioconversion, taking each conversional solution of Mylabris of I, II, III, IV group, 3500r/min is centrifuged 20min and removes PSB thalline, supernatant liquid is condensed into the mother liquid medicine containing Mylabris medical material 100mg/mL, and-20 DEG C of cold preservations are standby.

(5) anti tumor activity in vitro of each conversional solution of Mylabris compares

1. the preparation of cell reagent

1. cell culture fluid: in the RPMI1640 culture fluid of every 100mL containing 10% hyclone, 100U/mL penicillin and 100 μ g/mL streptomycins.

2. taking the dilution of appropriate amount of drug mother solution cell culture fluid and being configured to concentration is 1mg/mL pastille culture fluid, uses 0.22 μm Filter filtration sterilization, seal cold preservation standby, the used time according to required administration concentration cell culture fluid dilution allocates.

2. the Secondary Culture of cell

Take various cell cell culture fluid (containing 10% hyclone, 100U/mL penicillin and 100 μ g/mL streptomycins), It is placed in 37 DEG C, 5%CO₂Cell culture incubator in cultivate.Until cell grow 2～3 days to logarithmic (log) phase time, discard old culture fluid, use 0.25%Trypsin-EDTA trypsinization, is shown in that under microscope intercellular substance increases, kytoplasm retraction becomes round and i.e. terminates disappearing Change, remove pancreatin, add new culture fluid and blow and beat gently and make single cell suspension, Secondary Culture, passes on once for general 2～3 days.

The anti tumor activity in vitro of the most each conversional solution measures

According to different cell growth rates, adjust cell concentration 8～12 ×× 10⁴/ mL, every hole 100 μ L are inoculated in 96 holes Plate, after cultivation 24h makes it adherent, sops up old culture fluid, adds the culture fluid 100 μ L containing different pharmaceutical concentration, and arrange comparison Group (the not cell culture fluid of pastille)；Blank group (the most celliferous cell culture fluid), the often multiple hole of group design 4.It is placed in cell After incubator cultivates 48h, sopping up old culture fluid, add 100 culture fluid new for μ L and the CCK-8 reagent of 10 μ L, jog mixes.Continue Continuous cultivation reacts taking-up after 1h, according to CCK-8 kit method, with 630nm as reference wavelength, by BIO-RAD microplate reader at ripple Absorbance (OD) value is measured at long 450nm.Calculate

According to Principle of Statistics, experimental data withRepresenting, data acquisition SPSS 20.0 statistical software is carried out point Analysis, seeks half-inhibition concentration IC according to cell inhibitory rate IR statistical regression methods₅₀。

According to the growth speed situation of PSB in the conversional solution of Different Extraction Method, with each conversional solution to each tumor cell IC₅₀As index, consider the preferred Mylabris extracting method as conversion of substrate.

(6) experimental result of Mylabris extracting mode

1. the growing state of photosynthetic bacteria in different conversional solution

The growing state schematic diagram of photosynthetic bacteria in the different conversional solution that Fig. 1 provides for the present embodiment, can from Fig. 1 Going out, the growth adding membership suppression PSB of Mylabris, in I group, the growing multiplication speed of photosynthetic bacteria is very fast, and the growth cycle of PSB becomes Gesture is also closer to matched group.And the PSB poor growth in II group, III group, IV group, logarithmic (log) phase is delayed to the 3rd day.Point out Under identical Mylabris concentration, I group extracts the material of Mylabris to the embryotoxicity inhibitory action of photosynthetic bacteria than II group, III group and IV group Little.

2. the different conversional solution inhibitory action to hepatoma cell line BEL-7402

In the present embodiment, hepatoma cell line BEL-7402 inoculum density is 15 ××s 10⁴Individual/mL.

Suppression ratio that hepatoma cell line BEL-7402 is bred by each conversional solution of table 1 (n=4,)

As shown in Table 1, the suppression ratio of BEL-7402 cell is increased along with concentration and increases, in identical administration by each group medicine Under concentration, I group and III group are to the suppression ratio of cell relatively, and stronger than the inhibitory action of other two groups of medicines.SPSS Process and show that the half-inhibition concentration of BEL-7402 cell is followed successively by by I group, II group, III group, the medicine of IV group: 153.86 μ g/ mL；187.93μg/mL；160.05 μ g/mL, 262.08 μ g/mL.The suppression of hepatoma cell line BEL-7402 is imitated by prompting I group medicine The most best.

3. the different conversional solution inhibitions to HepG-2 cell line

In the present embodiment, HepG-2 cell line inoculum density is 10 ××s 10⁴Individual/mL.

Suppression ratio that HepG-2 cell line is bred by each conversional solution of table 2 (n=4,)

As shown in Table 2, under identical administration concentration, III group medicine is higher to the suppression ratio of HepG-2 cell line, calculates Obtain I group, II group, III group, the IV group IC to HepG-2 cell line₅₀It is 151.59 μ g/mL successively, 206.11 μ g/mL, 133.38μg/mL,289.98μg/mL；Prompting III group medicine is preferable to the inhibition of HepG-2 cell line.

4. the different conversional solution inhibitions to lung cancer A549 cell

In the present embodiment, lung cancer cell A-549 inoculum density is 8 ××s 10⁴Individual/mL.

Suppression ratio that lung cancer cell A-549 is bred by each conversional solution of table 3 (n=4,)

As shown in Table 3, each group medicine all has certain inhibitory action to lung cancer cell A-549, and with administration concentration in just Relevant.Under identical administration concentration, I group medicine is slightly high than other group medicines to the suppression ratio of cell.Be calculated I group, II group, III group, the IV group IC to HepG-2 cell line₅₀It is 390.56 μ g/mL successively, 611.19 μ g/mL, 425.09 μ g/mL, 495.85 μ g/mL, prompting I group medicine is good compared with what other were organized to the inhibition of lung cancer cell A-549.

5. medicine acts on the IC50 of each tumor cell

By experimental result, 1. to the inhibition of BEL-7402 cell by by force to weak: I group > > III group ＞ II group ＞ IV group； 2. to the inhibition of HepG-2 cell by by force to weak: III group ＞ I group ＞ II group ＞ IV group；3. the suppression to A-549 cell is imitated Fruit is by by force to weak: I group > > III group ＞ IV group ＞ II group.All in all the extracorporeal anti-tumor effect of Mylabris water extraction conversional solution is more Good, the PSB growth in I group is minimum by Mylabris toxicity inhibition, considers selection water extraction Mylabris in PSB conversion Mylabris and makees For the substrate (I group) converted.

Embodiment 2 Mylabris extracting solution bio-conversion process

(1) main material

Photosynthetic bacteria: the hydrogenlike silicon ion (Rhodobacter sphaeroides) that the red antibacterial of Purple Non-sulfur flora belongs to is purchased In Institute of Microorganism, Academia Sinica.

The culture medium of photosynthetic bacteria: sodium acetate 3.0g calcium chloride 0.05g dipotassium hydrogen phosphate 1.0g ammonium sulfate 1.0g yeast extract 2.0g magnesium sulfate 0.2g potassium dihydrogen phosphate 0.7g liquid microelement 5ml water 1000ml.

Trace element solution: disodiumedetate 2.000g green vitriol 0.200g boric acid 1.500g sulphuric acid Copper 0.030g bis-molybdic acid hydrate sodium 0.450g anhydrous manganese 1.200g Zinc vitriol 0.150g Nickel dichloride hexahydrate 0.020g water 1000ml.

(2) key instrument

Shimadzu UV2450 ultraviolet-uisible spectrophotometer (Shimadzu Corporation of Japan)；Growth cabinet (the south of the River, Ningbo instrument Factory)；BL-220H electronic balance (SHIMADZU company of Japan)；DSX-280B Portable pressure steam sterilizing pot (Shen, Shanghai peace)； SW-CJ-1F clean bench (SuZhou Antai Air Tech Co., Ltd.)；Liquid-transfering gun (Eppendorf company).

(3) method and result

1. the growth curve of photosynthetic bacteria

Preparation photosynthetic bacteria culture medium, the photosynthetic bacteria being in logarithmic (log) phase of inoculation 10% (volume ratio), if blank group and Dosing group (experimental group), often 4 Duplicate Samples of group, the adding consistency of dosing group is 10mg/mL (amount of Mylabris is calculated by crude drug amount), from And measure the growth curve of photosynthetic bacteria in the case of not dosing and dosing.Within every 12 hours, measure bacterium solution extinction under 660nm Degree, measures 12 days altogether.Corresponding growth curve is shown in accompanying drawing 2.

There is not the laundering period in blank group as can be seen from Figure 2, and dosing group exists the obvious laundering period；96th hour blank group Terminating logarithmic (log) phase, enter stable phase, absorbance is stable at about 5.For dosing group, within the 36th hour, starting by the 96th hour In obvious logarithmic (log) phase, after 96 hours, amplification slows down；It is longer to there is the time in blank group stable phase, within the 8th day, enters decline phase. There is not obvious stable phase in dosing group, but same 8th day enters decline phase；Determine that the time of bioconversion is 4-～accords with for 8 days Close the feature in photosynthetic bacterium growth cycle.On the one hand, photosynthetic bacterium growth is vigorous in this process, and enzyme is lived bigger, converts energy Power is higher；On the other hand, after entering decline phase, photosynthetic bacteria conversion refuse increases, and is unfavorable for bioconversion post analysis.

2. the investigation of Mylabris safe concentration

Accurately weigh appropriate Mylabris coarse powder and add the water soaking 30min of 20 times amount, heating and refluxing extraction 2h, cool down, filter, Obtain Mylabris Aqueous extracts.

With PSB culture medium be made into successively the medical material concentration containing Mylabris be 0,4,8,10,12,16, the solution of 20mg/ml, train Support the PSB being in logarithmic (log) phase to inoculate with 10% inoculum concentration (volume ratio), be placed on Anaerobic culturel in illumination box, every 12h is separately sampled, surveys the OD value of PSB to represent the growth of PSB, METHOD FOR CONTINUOUS DETERMINATION 96h at 660nm.Parallel 3 of each concentration Sample, averages.With OD_n/OD₀(OD₀It is the initial optical density of solution, OD_nIt is the optical density of n hour) reaction photosynthetic bacteria Growing state.

Figure of description 3 is the growing state of photosynthetic bacteria under different Mylabris extracting solution mass concentration, can by description Fig. 3 So that to find out Mylabris drug level be 0,4,8,10mg/mL time, PSB growth tendency is substantially the same, and concentration be 12,16,20mg/mL Time PSB bacterial growth slow, fast growing period postpones or does not occurs.In the liquid of visible conversion, the concentration of Mylabris is the biggest grows PSB Inhibitory action is the most obvious.

Utilize OD value and the OD value ratio OD of 0h of each solution 96h recorded_96h/OD_0hFor index (such as table 4), lead to SPSS Whether the matched group crossing average minimum difference method (LSD) relatively each Mylabris concentration and 0mg/mL has significant difference (P ＜ 0.05), Determine the safe concentration that PSB is grown by Mylabris.

The OD of the different Mylabris strength solution of table 4_96h/OD_0h

As can be seen from Table 4 along with the increase of Mylabris concentration, the cell concentration change ratio OD of photosynthetic bacteria_96h/OD_0hGradually Reduce, illustrate that the growth inhibited of PSB is increased along with concentration and strengthens by Mylabris.Find out from P value, Mylabris concentration is 4,8,10mg/ The conversion group of mL and matched group do not have significant difference (P ＞ 0.05), it was demonstrated that concentration 4,8, under the concentration of 10mg/mL to photosynthetic The growth effect of antibacterial and matched group do not have significant difference.Therefore the safe concentration that PSB converts Mylabris is 4～10mg/mL, is more than 10mg/mL will produce obvious toxic action, the growth of suppression PSB to PSB.Therefore Mylabris during photosynthetic bacteria bioconversion Mylabris Drug level be 4～10mg/mL.

Toxicity before and after Mylabris extracting solution bioconversion is compared by embodiment 3 photosynthetic bacteria

1. main material

Mylabris is the dry polypide of Meloidae insecticide Mylabris phalarata Mylabris phalerata Pallas, is purchased from peace Fructus Liquidambaris prepared slices of Chinese crude drugs company limited of state；Mylabris Aqueous extracts and the self-control of Mylabris conversional solution equal laboratory.

2. animal subject

Animal subject: SPF level Kunming kind healthy mice (18～22g), male and female half and half, is purchased from Traditional Chinese Medicine University Of Guangzhou real Test animal center, SCXK (Guangdong) 2013-0020.

3. method and result

(1) preparation of medicine

Accurately weigh Mylabris coarse powder (30 mesh, 50 DEG C are dried 6 hours) 4 parts, add the water of 20 times amount, soak 30min, heat back Stream extracts 2h, cools down, filters, obtains extracting solution.

Extracting solution is evaporated to the 5% of conversional solution cumulative volume, the photosynthetic bacteria of inoculation 10% (volume ratio) logarithmic (log) phase (PSB), obtain calculating the concentration conversional solution as 10mg/mL with Mylabris medical material.Anaerobic culturel, bioconversion 8 days.Take Mylabris to convert Liquid, 3500r/min is centrifuged 20min and removes PSB thalline, and supernatant liquid is condensed into the mother liquid medicine containing Mylabris medical material 100mg/mL ,- 20 DEG C of cold preservations are standby.Mouse stomach uses administration concentration needed for distilled water furnishing before being administered.

(2) acute toxicity testing

1. SPF level Kunming mouse, random packet, often group 10, male and female half and half are taken.According to preliminary result Mylabris metabolism The whole lethal minimum dose of liquid is Dmin1620mg/kg and whole non-lethal maximal dose Dmax is 3000mg/kg, will Mylabris Aqueous extracts and Mylabris conversional solution are respectively with various dose gastric infusion, and medicine is respectively provided with 6 dosage groups.Matched group The distilled water of gavage same volume.Before mice is administered, water 12h, gavage volume 0.2mL/10g are can't help in fasting.To observation post administration mouse Toxic reaction and death condition, Continuous Observation 14 days, every body weight weighing survival mice for 2 days.According to various dose group mice Dead calculating LD₅₀。

2. mouse liver and Pathological sections observation

Take the mice of high, normal, basic three dosage groups, put to death when dead mouse and at the end of experiment in 14 days, dissect little immediately Mus takes out liver, kidney, makes paraffin section, and under microscope, (100 times) are observed and taken pictures

(3) result

1. mouse toxicity reaction symptom and time of occurrence

Mylabris Aqueous extracts is similar with the toxic reaction symptom of Mylabris conversional solution, and the biggest toxic reaction of dosage is the strongest, appearance Time is also the most early.Mylabris Aqueous extracts group and Mylabris conversional solution group all occur that spontaneous activity in mice reduces after being administered, and extremity are crouched down, abdomen Portion becomes big, bradykinesia, sexual organ's hyperemia enlargement；Eye closing, refusing to eat, urinary incontinence；Rapid breathing before dead, twitch, shed tears.Mylabris water Extract group poisoning time of occurrence concentrates upon administration 2～3h.Survival mice alleviates more than 4～5 days symptoms, and just gradually recovers Often.Mylabris conversional solution group poisoning time of occurrence concentrates upon administration 4～6h, survival mice 2～gradually recover normal after 3 days.

2. the relation of mortality rate and dosage and reaction

Various dose group mice accumulation death toll is shown in Table 5 with the relation of dosage

Dead mouse situation and LD50 before and after the conversion of table 5 Mylabris

The dosage of Mylabris Aqueous extracts and Mylabris conversional solution is the highest as can be seen from Table 5, and mouse death rate is the highest, toxicity The biggest.The homing method using SPSS20 tries to achieve Mylabris Aqueous extracts LD₅₀=206mg/kg, 95% credible is limited to: 179～233mg/ kg；And Mylabris conversional solution LD₅₀=2036mg/kg, 95% fiducial limit: 1923～2150mg/kg.LD₅₀It is worth the biggest then drug toxicity The least, thus prove that the toxicity that mouse stomach is administered after photosynthetic bacteria converts by Mylabris significantly reduces, from the point of view of quantitative relation, Mylabris toxicity after photosynthetic bacteria converts probably reduces by 10 times.

3. Anatomical Observation

The mice of different dosing dosage group is dissected, finds that esophagus, gastric mucosa fester hemorrhage；Stomach foaming edema, stomach Intestinal is congested, splanchnocoel hemorrhagic effusion；There is red macules on pulmonary edema and surface, and heart color and luster is dim；Liver edge cloudy swelling, color and luster Dim；Kidney kermesinus, muddy without transparent feel.

4. mouse liver and Pathological sections observation result

The lobules of liver structural integrity of the mouse liver tissue of matched group, shows as hepatocyte centered by central vein in putting Penetrating shape arrangement, portal area is without inflammatory cell infiltration and substantially proliferation of fibrous tissue, and hepatocyte is completely normal, and nucleus is oval, Can clearly see kernel.Administration group there will be hepatocyte fall into disarray in various degree, inflammatory cell infiltration, lobules of liver structure quilt The situations such as destruction.

The kidney renal tubules of control group mice is normal, and glomerule is circular complete, and the Mouse Kidney influential point of viscera selectivity of poisoning occurs The situations such as glomerule pyknosis, volume increase, renal tubular necrosis.

From the point of view of the result of mouse toxicity reaction symptom and time of occurrence, the mice poisoning time of occurrence collection of Mylabris Aqueous extracts In upon administration 2～3h, survival mice alleviates more than 4～5 days symptoms, and gradually recovers normal.And the mice of Mylabris conversional solution Poisoning time of occurrence concentrates upon administration 4～6h, survival mice 2～gradually recover normal after 3 days.The time that poisoning occurs prolongs After, recover normal time in advance, prompting toxicity reduces.Median lethal dose(LD 50) LD of Mylabris conversional solution₅₀For 1383mg/kg, Mylabris water Extract LD₅₀For 206mg/kg, it was demonstrated that Mylabris toxicity after photosynthetic bacteria converts significantly reduces, and has reached the effect of attenuation.

Mylabris is obvious to the hepatic tissue Attenuation of mice before and after converting, and pathology is mainly manifested in central vein congestion, scorching Cellular infiltration, lobules of liver is destructurized, local necrosis.Pathological toxicity is inconspicuous, mainly cashes at slight glomerule Increase, renal tubular cell pathological changes；Being likely due in the case of single-dose, kidney is not produced by toxic component Organic damage.But anatomic observation has congested point, thoracic cavity to have a hematocele to heart adventitia, prompting chmice acute is poisoned to death may be with Blood circulation is relevant.

The outer tumor-inhibiting action test of embodiment 4 Mylabris conversion solution

1. medicine and reagent

Mylabris is the dry polypide of Meloidae insecticide Mylabris phalarata Mylabris phalerata Pallas, is purchased from peace Fructus Liquidambaris prepared slices of Chinese crude drugs company limited of state；Hyclone (Ilex purpurea Hassk.[I.chinensis Sims)；RPMI1640 culture fluid (Hyclone)；0.25% Trypsin-EDTA(GIBCO)；CCK-8 test kit (green skies company)；PBS(GIBCO).

2. cell strain

3. key instrument

GalaxyS+ CO2 gas incubator (Hua Yuehang Instrument Ltd.)；Chongqing Austria spy's inverted biological microscope BDS200；BIO-RAD Model-1680 microplate reader；0.22 μm filter (U.S. Pall).

4. method and result

(1) preparation of cell reagent

Taking the dilution of appropriate Mylabris conversional solution cell culture fluid and being configured to concentration is 1mg/mL pastille culture fluid, uses 0.22 μm Filter filtration sterilization, seal cold preservation standby.Used time is according to experiment needs, dilution allotment.

(2) recovery of cell, cultivation

The recovery of cell: conventional sterilant adds the culture fluid of 10-15mL in culture bottle, quickly takes out swollen from liquid nitrogen is shone The cryovial of tumor cell strain, and put into rapidly in the water-bath of warmed-up 37 DEG C, constantly shake, make cryopreservation tube melt rapidly (in 1min)；Move to frozen stock solution add in cell culture fluid, within second day, change a culture fluid, continue to cultivate.

The amplification culture of cell: conventional treatment, adds 500ul 0.25% trypsin and digests attached cell, in Microscope observes the degree determining digestion, is passed on according to the ratio of 1:2 to 1:4 by cell, is placed in CO2 incubator and trains Support.When, growth rate good until cell state is fast, cell is carried out frozen and tests.

(3) inhibitory action of tetrazolium salts (MTT) colorimetric determination various tumor cell strains growth

1. CCK-8 method is with 630nm as reference wavelength, measures absorbance (OD) by BIO-RAD microplate reader at wavelength 450nm Value.Calculate cell inhibitory rate IR (%)=(matched group experimental group OD)/(matched group blank group) 100%.

2. the suppression ratio of extracorporeal anti-tumor measures

Human liver cancer cell HepG-2, human liver cancer cell SMCC-7211, human lung cancer cell A549, people uterine cancer cells Hela With 8 × 10⁴Individual/mL carries out kind of a plate as inoculum concentration；Human liver cancer cell QGY-7701 is with 5 × 10⁴Individual/mL is carried out as inoculum concentration Plant plate.

(4) result of the test

1. the medicine inhibitory action to HepG-2 cell line

Suppression ratio/% that HepG-2 cell line is bred by table 6 medicine (n=4,)

Table 7 medicine is to the regression equation of HepG-2 cytosis and middle effect concentration

Knowable to table 6, table 7, Mylabris conversional solution is inhibited to HepG-2 cell, during doses, suppression ratio with The prolongation of action time and rise, under certain time, suppression ratio rises along with the rising of drug level, in obvious agent Amount-time dependence.Process through SPSS 22.0 and draw the Mylabris conversional solution IC to 24h, 48h, 74h of HepG-2 cell₅₀Depend on Secondary it is: 249.44 μ g/mL (confidence interval is (214.37-322.23 μ g/mL))；(confidence interval is 168.11 μ g/mL (155.00-183.87μg/mL))；106.60 μ g/mL (confidence interval is (68.13-131.68 μ g/mL)).

2. the medicine inhibitory action to uterus carcinoma Hela cell

The each conversional solution of table 8 to the suppression ratio/% of uterus carcinoma Hela cell proliferation (n=4,)

Table 9 medicine is to the regression equation of Hela cytosis and middle effect concentration

	Regression equation	IC<sub>50</sub>	Fiducial range
				24h	Y=-4.88X+1.837	452.79	343.89-545.32
48h	Y=-5.343X+2.184	279.76	242.90-351.53
				72h	Y=-6.613X+2.933	126.19	108.43-140.70

From table 8,9, Mylabris conversional solution is inhibited to Hela cell, and during doses, suppression ratio is along with work Rising with the prolongation of time, under certain time, suppression ratio rises along with the rising of drug level, in obvious dosage-time Between dependency.Process through SPSS 22.0 and draw the Mylabris conversional solution IC to 24h, 48h, 74h of Hela cell₅₀It is followed successively by: 452.79 μ g/mL (confidence interval is (343.89-545.32 μ g/mL))；(confidence interval is (242.90-to 279.76 μ g/mL 351.53μg/mL))；126.19 μ g/mL (confidence interval is (108.43-140.70 μ g/mL)).

3. the medicine inhibition to lung cancer A549 cell

The each conversional solution of table 10 to the suppression ratio of hepatocarcinoma A549 cell proliferation (n=4,)

Table 11 medicine is to the regression equation of A549 cytosis and middle effect concentration

From table 10,11, Mylabris conversional solution is inhibited to A549 cell, during doses, suppression ratio along with The prolongation of action time and rise, under certain time, suppression ratio rises along with the rising of drug level, in obvious dosage- Time dependence.Process through SPSS 22.0 and draw the Mylabris conversional solution IC to 24h, 48h, 74h of A549 cell₅₀It is followed successively by: 249.44 μ g/mL (confidence interval is (214.37-322.23 μ g/mL))；(confidence interval is (98.39-to 112.36 μ g/mL 126.48μg/mL))；66.82 μ g/mL (confidence interval is (53.37-78.50 μ g/mL)).

4. the medicine inhibitory action to hepatocarcinoma QGY-7701 cell

The each conversional solution of table 12 to the suppression ratio/% of hepatocarcinoma QGY-7701 cell proliferation (n=4,)

Table 13 medicine is to the regression equation of QGY-7701 cytosis and middle effect concentration

Knowable to table 12, table 13, Mylabris conversional solution is inhibited to QGY-7701 cell, during doses, and suppression Rate rises along with the prolongation of action time, and under certain time, suppression ratio rises along with the rising of drug level, in significantly Dosage-time dependence.Process through SPSS 22.0 and draw the Mylabris conversional solution IC to 24h, 48h, 74h of QGY-7701 cell₅₀ It is followed successively by: 234.80 μ g/mL (confidence interval is (208.44-280.94 μ g/mL))；(confidence interval is 165.98 μ g/mL (151.85-183.18μg/mL))；(confidence interval is (84.44-105.12 μ g/mL) to 95.68 μ g/mL.

5. the medicine inhibition to pulmonary carcinoma SMC-7211 cell

The each conversional solution of table 14 to the suppression ratio of hepatocarcinoma SMC-7211 cell proliferation (n=4,)

Table 15 medicine is to the regression equation of SMC-7211 cytosis and middle effect concentration

From table 14,15, Mylabris conversional solution is inhibited to SMC-7211 cell, during doses, and suppression ratio Rising along with the prolongation of action time, under certain time, suppression ratio rises along with the rising of drug level, in obvious agent Amount-time dependence.Process through SPSS 22.0 and draw the Mylabris conversional solution IC to 24h, 48h, 74h of SMC-7211 cell₅₀Depend on Secondary it is: 289.30 μ g/mL (confidence interval is (260.54-336.36 μ g/mL))；(confidence interval is 207.45 μ g/mL (189.78-228.95μg/mL))；119.60 μ g/mL (confidence interval is (105.54-131.40 μ g/mL)).

(5) cellular morphology is observed

From the point of view of cell inhibitory rate, the medicine suppression ratio A549 > QGY-7701 > HepG-2 > SMC-7211 to cell 48h > Hela, selects the cell of suppression ratio first three to carry out cellular morphology under mirror and observes, i.e. medicine to hepatocarcinoma QGY-7701, A549, Cellular morphology after HepG-2 cell 48h effect is observed.

QGY-7701, A549, HepG-2 blank group, tumor cell adherent growth, closely, polygon presents in arrangement, After medicine effect, tumor cell shrinkage becomes round or dead, Microscopic observation cell concentration bad also less than the cell attachment of blank group and Swimming in above, have fragment shape, along with administration concentration is the highest, floating the most of cell death shrinkage, these phenomenons show Mylabris Conversional solution can substantially suppress the growth of tumor cell after acting on cell.It is also seen that along with the increasing of drug level from figure Adding, A549 cell occurs that the cell concentration become apparent from reduces, and tumor cell shrinkage becomes round the phenomenon of death, by the figure of Microscopic observation Sheet will also realize that Mylabris conversional solution is obvious to inhibitory action relatively other cells of A549 cell, and inhibitory action is higher.

The present embodiment has investigated Mylabris conversional solution to the timeliness of various tumor cell strains and dose-effect, i.e. uses the inspection of CCK-8 method Survey various dose, the effect different time sensitivity to cancerous cell, carry out the Primary Study inhibitory action to tumor.Experiment in vitro is made For the primary dcreening operation of antitumor drug, it it is the basis of experiments in vivo.

Knowable to the result of experiment, different Mylabris conversional solution concentration is equal in different action time to different cell strains There is inhibitory action, and present obvious time and dose dependent.

In the problem of laboratory early stage is tested, showing composition Study before and after converting, Mylabris is transformed through photosynthetic bacteria Total cantharidin content in journey is declining, the external anti-tumor experiment to various kinds of cell of the Mylabris conversional solution after process optimization The antitumor action of prompting Mylabris conversional solution strengthens, and the prompting of this result may produce antitumous effect after converting and be better than cantharidin New chemical composition.

The effect to transplanted hepatoma H22 of the embodiment 5 Mylabris conversional solution

1. strain and animal

Tumor strain: Murine ascites hepatocarcinoma cell strain H22, is given by basis teaching and research room of Guangdong Pharmaceutical University.

Animal: kunming mice, male and female half and half, body weight (20 ± 2) g, purchased from Traditional Chinese Medicine University Of Guangzhou experimental center, the quality certification Number: SCXK (Soviet Union) 2008-0010.

2. main material and instrument (omiting)

3 methods and result

(1) rat liver cancer model is set up

First routine carries out the recovery of H22 tumor cell line and passes on, and carries out three times altogether and passes on.Adjust cell number 1.0 × 10⁷/ mL, the cell viability of tumor, 95%, is inoculated in the right axil of mice by every Mus 0.2mL subcutaneous, is prepared as solid tumor animal mould Type；Separately it is inoculated in mouse peritoneal with every Mus 0.2mL, is prepared as Animals with ascitic tumors model.

(2) experiment packet and administration

1. the conversion of Mylabris conversional solution dosage

Through preliminary experiment, set the dosage of Mylabris conversional solution as 100mg/kg, 200mg/kg, 300mg/kg.Kunming mouse Gavage capacity be 0.2mL/10g, the concentration of the Mylabris conversional solution of the lowest middle high group is 5mg/mL, 10mg/mL, 15mg/mL.

2. Mylabris Aqueous extracts dosage

The dosage of Mylabris Aqueous extracts be the low dosage 100mg/kg of Mylabris conversional solution, i.e. concentration be 5mg/mL.

3. 5-Fu collocation method

Every day, the dosage of 5-Fu was 15mg/kg, adds normal saline and is made into 0.75mg/mL, standby.

4. packet is tested

After inoculation 24h, Random Design packet, is divided into 6 groups, often group 10.I.e. blank group, lotus tumor model group, positive right According to (the dosage in BMDX of dosage group in group (5-Fu group), Mylabris conversional solution low dose group (BMDX low dose group), Mylabris conversional solution Group) and Mylabris conversional solution high dose group (BMDX high dose group),.

5. administration processes

Kunming mouse is fed in Guangdong Pharmaceutical University's animal center.After inoculated tumour cell 24h, start gavage and abdominal cavity note Penetrate, every day 1 time, continuous 10 days.

Normal group, model group are all with 0.9%NaCl solution gavage 0.4ml/20g/d；5-Fu group gives 5-Fu solution abdominal cavity note Penetrate 15mg/kg/d；Give and basic, normal, high various dose Mylabris conversional solution gavage respectively, by 0.2mL/10g/ for 4th group to the 6th group D is carried out.

(3) ascites tumor observation index and detection method

1. the observation of general status

Experimentation is observed Kunming mouse mental status situation, autonomic activities situation, diet situation, feces situation, build Visual condition, the time that ascites occurs and hair situation etc..

2. the mensuration of ascites volume

Putting to death mice after medication in the tenth day terminates 24h, extraction ascites also measures ascites volume.

Respectively organize the observation of survival time of mice

After being administered 10d, stopping being administered, stop to 20d, record, Kaplan-Meier method draws curve life cycle.

(4) solid tumor observation index and detection method

1. the observation of general status

Experimentation is observed Kunming mouse mental status situation, autonomic activities situation, diet situation, feces situation, build Visual condition and hair situation etc..

2. tumor weight measures

Claim animal weight, put to death, open oxter, peel off tumor body, weigh, record.

3. thymus coefficient and Spleen coefficient

After experimental administration terminates, put to death mice, take out thymus and spleen, calculate spleen index and thymus index.

(5) ascites tumor each index result

1. the observation of general status

Relatively model group, positive controls (5-Fu group), BMDX dosage group low, middle all make the mental status situation of mice have Being improved, wherein the situation of 5-Fu group mice is preferable, along with the rising of the dosage of Mylabris conversional solution, the mice of BMDX high dose group State is not good enough, occurs in that the phenomenon of dead mouse.

2. the mensuration of ascites volume is shown in Table 16

Table 16 respectively organize mouse ascites amount observation (n=10,)

Group	Dosage mg/kg	Ascites tumor
			Matched group	NS	——
Model group	NS	11.54±2.52
			5-Fu group	15	0.00±0.00**
Mylabris Aqueous extracts group	100	13.2±1.82**
			BMDX low dose group	100	12.8±2.64**
Dosage group in BMDX	200	11.8±3.27
			BMDX high dose group	300	11.6±2.54

Note: compare * P < 0.05, * * P < 0.01 with model group

Relatively model group, 5-Fu group does not have the phenomenon of ascites, the i.e. treatment to ascites tumor during experimental administration Effective；

The ascites volume that the mice of Mylabris Aqueous extracts group produces is more than model group, i.e. to the ascites volume of mice without improving Effect；

Mylabris conversional solution basic, normal, high dosage group after metabolism to the therapeutic effect of the ascites tumor of mice along with medicine The increase of concentration, therapeutic effect also increases, but poor effect.

The observation respectively organizing survival time of mice is shown in Table 17

The average life of table 17 mice

Note: compare * P < 0.05 with model group；**P<0.01

There is not the phenomena of mortality of mice, relatively mould in the mice of 5-Fu group mice upon administration mice when 20d stops experiment Type group, 5-Fu protracted drug life-span of mice.

Mylabris conversional solution dosage group low, middle relatively model group, all extends the life span of mice, compares with model group and have Significant differences (* * P < 0.01)；The mice of high dose group, when administration time is also not up to 10d, occurs in that dead mouse Phenomenon.

(6) solid tumor each index result

1. the observation (omiting) of general status

2. tumor weight measurement result is shown in Table 18.

Table 18 respectively organize mice tumour inhibiting rate observation (n=10,)

Note: compare * P < 0.05 with model group；**P<0.01

Compare with model group: tumor is had and obviously presses down by Mylabris Aqueous extracts, Mylabris conversional solution basic, normal, high dosage group Make and use (* * P < 0.01), but tumor killing effect is not as 5-Fu group.

3. thymus coefficient and Spleen coefficient the results are shown in Table 19

Table 19 respectively organize mouse thymus and spleen coefficient mensuration (n=10,)

Note: compared with model group: * * P < 0.01, * P < 0.05；Contrast with positive group:^##P<0.01、^#P<0.05

Compared with Mylabris Aqueous extracts:^△△P<0.01、^△P<0.05

Compared with model group, in BMDX, the spleen coefficient of dosage group and model group have significant differences (* * P < 0.01), Its thymus coefficient also has significant difference (* P < 0.05) compared with model group.

Contrasting with 5-Fu group, BMDX low dose group, the thymus of middle dosage group and spleen coefficient all have very significant poor with it Different (^##P<0.01)。

Compared with Mylabris Aqueous extracts, BMDX low dose group, the thymus of middle dosage group and spleen coefficient all have highly significant with it Sex differernce (^△△P<0.01)。

Understanding in this experimentation, Mylabris Aqueous extracts and Mylabris conversional solution same dose are to the therapeutic effect of ascites and mould Type model group ratio has the difference (* * P < 0.01) of highly significant, and conversional solution is along with the increase of drug level, and therapeutic effect is on the contrary Decline.

Compared with matched group, Mylabris conversional solution low dose group, middle dosage group all extend the life cycle of H22 ascites tumor mice, Improve life time, difference has the meaning of highly significant.

Mylabris conversional solution tumour inhibiting rate is 18.11%, 25.98%, 37.01%, and Mylabris Aqueous extracts tumour inhibiting rate is 26.7%, right The H22 tumor of mice is inhibited, and relatively model group has the raising (P < 0.01) of very significant, and positive drug tumour inhibiting rate is better than Mylabris.And positive drug and Mylabris Aqueous extracts are nonsensical to the effect of immunologic function, Mylabris conversional solution but can strengthen by highly significant The immunologic function of mice, it is thus possible to play antitumor action well.

The effect to Lewis lung carcinoma Lewis of the embodiment 6 Mylabris conversional solution

1. strain and animal

Tumor strain: murine lung cancer cell strain Lewis, purchased from Chinese Academy of Sciences's cell bank.

Animal: C57 mice, male and female half and half, 6-8 week old, body weight (18 ± 2) g, purchased from Guangdong Province's animal center, the quality certification Number: SCXK (Guangdong) 2015-0020.

2. main material is with instrument ibid.

3. method and result

(1) Lewis mice lung cancer model is set up

First routine carries out the recovery of Lewis tumor cell line and passes on.Ready Cell sap is connect by every Mus 0.2mL Plant subcutaneous in the right axil of mice, be prepared as solid tumor animal model.

(2) experiment packet and administration

The operational approach of this step is with the experiment packet of hepatocarcinoma in embodiment 5 and medication.

(3) solid tumor observation index and detection method

1. the observation of general status: mental status situation, autonomic activities situation, diet situation, feces situation, build outward appearance The detection of situation, body weight and hair situation etc..

Body weight rate of increase=((body weight before body weight-administration after administration)/it is administered front body weight) × 100%.

2. tumor weight measures.

3. thymus coefficient and the mensuration of Spleen coefficient: be administered after terminating, put to death mice, take out thymus and spleen, calculates spleen and refers to Number and thymus index.

(4) each index of solid tumor the results are shown in Table 20:

Table 20 respectively organize mouse thymus, spleen index mensuration (n=10,)

Note: compared with model group: * * P < 0.01, * P < 0.05；Contrast with positive group:^##P<0.01、^#P<0.05；With Mylabris Aqueous extracts group is compared:^△△P<0.01、 ^△P<0.05

1. the observation of general status

Compared with model group, the weight gain of the mice of administration group has reduced；Compared with 5-Fu group, Mylabris water carries The weight gain highly significant of liquid and Mylabris conversional solution improve (^##P<0.01)；Compared with Mylabris Aqueous extracts, Mylabris conversional solution The raising of weight gain highly significant (^△△P<0.01)；The mice of Mylabris conversional solution group is along with the increase of mice dosage, little The weight gain of Mus is gradually lowered.

2. tumor weight measures

Positive drug 5-Fu is big to the suppression ratio of tumor, and Mylabris conversional solution suppression ratio is along with the increase of dosage, suppression Rate strengthen, compared with Mylabris Aqueous extracts group mice, have highly significant diversity (^△△P<0.01)。

Mylabris conversional solution basic, normal, high dosage tumour inhibiting rate is followed successively by 21.47%, 27.72%, 33.39%, middle and high dosage group Mice the suppression ratio of tumor is better than the 25.33% of Mylabris Aqueous extracts, there is the raising (P < 0.01) of very significant, positive Medicine tumour inhibiting rate 49.29%.

3. thymus coefficient and Spleen coefficient

Contrast with positive drug 5-Fu group, Mylabris Aqueous extracts, BMDX low dose group, middle dosage group, the thymus of high dose group All have with it with spleen coefficient significant differences (^##P<0.01)；Compared with Mylabris Aqueous extracts, the Mylabris conversional solution spleen to mice Dirty coefficient, thymus coefficient all have very significant increase (^△△<0.01)。

To sum up result shows: the effective of tumor is treated in positive drug local, but the best to body mass action；Mylabris turns The advantage changing liquid treatment entity tumor is to regulate very significantly and enhancing human body immunity function.Thymus and spleen are important Immune organ, thymus coefficient has reacted quantity and the function of immune cell to a certain extent, and spleen coefficient is the most to a certain degree Reflecting the immunologic function of body, body's immunity imbalance often results in the generation of tumor, and immunologic function is suppressed causes tumor Cell persistent hyperplastic.Solid tumor models in this experiment, the thymus of mice, spleen coefficient relatively positive drug 5-Fu, Mylabris water carry Liquid all has very significant to increase, and therefore Mylabris conversional solution medicine may play antitumor by regulating and strengthening immunologic function Effect.

Claims

1. the application of photosynthetic bacteria attenuation synergistic in Mylabris anticancer function, described photosynthetic bacteria is that Purple Non-sulfur flora is red carefully The hydrogenlike silicon ion (Rhodobacter sphaeroides) of Pseudomonas.

Apply the most according to claim 1, it is characterised in that photosynthetic bacteria has anti-lung cancer, hepatocarcinoma, uterus carcinoma effect Application in Mylabris conversional solution.

Apply the most according to claim 2, it is characterised in that photosynthetic bacteria hepatoma cell strain HepG-2, BEL-7402, Application in SMCC-7211, QGY-7701；Photosynthetic bacteria application in lung cancer cell types；Photosynthetic bacteria is at uterus carcinoma Application in cell strain Hela.

4. the preparation method of a Mylabris conversional solution with anticancer function, it is characterised in that described preparation method specifically includes Following steps:

S2: concentrated extracting solution also inoculates photosynthetic bacteria described in claim 1, is subsequently adding PSB culture medium and carries out bioconversion；

Preparation method the most according to claim 4, it is characterised in that described photosynthetic bacteria be in logarithmic (log) phase photosynthetic carefully Bacterium, the inoculum concentration of described photosynthetic bacteria is 7～10%.

Preparation method the most according to claim 4, it is characterised in that the time of the bioconversion in described step S2 be 8～ 12 days.

Preparation method the most according to claim 4, it is characterised in that the mass concentration of Mylabris extracting solution described in step S1 is 7～10mg/mL.

Method the most according to claim 4, it is characterised in that the extracting solution in step S1 is water；Concentration extraction in step S2 The volume of liquid is to the 5～8% of conversional solution cumulative volume；The pH of described conversional solution system is 6～9.

9. the Mylabris conversional solution that method described in claim 4～8 prepares.

10. a Mylabris biological preparation with anticancer function, it is characterised in that described Mylabris biological preparation contains claim Mylabris conversional solution described in 9.