CN102688280A - Preparation method for radiation-resistant Chinese medicine fermentation liquor - Google Patents

Abstract

The invention provides a preparation method of anti-radiation traditional Chinese medicine fermentation liquid, relates to the field of traditional Chinese medicine processing and biological fermentation, in particular to the fermentation method of traditional Chinese medicines astragalus, notoginseng and green tea. In the invention, edible and medicinal fungi are used as starting strains of fermentation to respectively ferment three kinds of traditional Chinese medicines, and carry out dosage proportioning combination of the three kinds of fermented liquids obtained by the method. Animal experiments show that the traditional Chinese medicine fermentation liquid of the present invention has obvious anti-radiation effect. The present invention is characterized in that the traditional Chinese medicine fermented liquid can be directly used as food (drink) products, health care products and medicines, or used as raw materials for preparing food (drink) products, health care products and medicines, and has good performance in preventing and treating radiation damage. development and application prospects.

Description

Preparation of an anti-radiation traditional Chinese medicine fermentation liquid

Technical field:

The invention relates to the field of traditional Chinese medicine processing and biological fermentation, in particular to the fermentation process of traditional Chinese medicines astragalus, notoginseng and green tea, and the anti-radiation effect of the fermentation products obtained by the method.

Background technique:

Total flavonoids of Radix Astragali is one of the main active ingredients of Radix Astragali, there are more than 30 kinds, and its anti-radiation effect is obvious. Yang Yingxue and others studied the protection of total flavonoids of Astragalus on radiation-damaged mice. The results showed that the total flavonoids of Astragalus had obvious anti-radiation effect, the survival rate of irradiated mice was significantly improved, and the damage of peripheral blood WBC, PLT and spleen was significantly reduced ^[1] . Xu Zhengmin et al. took human normal bone marrow mesenchymal stem cells as the research object and irradiated them with ⁶⁰ Coγ-rays. The results showed that the total flavonoids of astragalus can play a role in radiation protection of human normal bone marrow mesenchymal stem cells ^[2] .

Saponins widely exist in many traditional Chinese medicines, and the content of total saponins in Panax notoginseng is relatively large, and in recent years some scholars have found that total saponins of Panax notoginseng has a strong anti-radiation effect. In order to explore the anti-radiation efficacy of Panax notoginseng saponins, Chen Xiwen et al. conducted experimental research. The results showed that Panax notoginseng saponins could significantly increase the number of red blood cells, white blood cells and bone marrow nucleated cells in the irradiated mice, and the thymus and spleen indexes of the mice were significantly improved. Increased, the carbon clearance index increased ^[3] . Zou Dan et al. found that Panax notoginseng saponins can significantly increase the total number of peripheral blood WBC, Hb, BMC and the yields of CFU-S and CFU-GM colonies in irradiated mice, and concluded that Panax notoginseng saponins can enhance bone marrow hematopoiesis. Function, to achieve antagonism to the bone marrow suppression in mice after radiation ^[4] .

Tea polyphenols are the most valuable functional components in tea, with the characteristics of wide distribution and high content, mainly including catechins, anthocyanins, flavonoids and flavonols, phenolic acids and phenolic acids ^[5] . In recent years, many scholars have studied the medical and health effects of tea polyphenols, and found that tea polyphenols have various activities such as scavenging free radicals and anti-aging, and confirmed that they have obvious anti-radiation effects ^[6,7] . Cao Mingfu studied the damage protection of tea polyphenols on mice irradiated by ⁶⁰ Co γ through experiments. The results showed that tea polyphenols could improve the immune organ index, blood picture, CFU-S and granulocyte MI of irradiated mice, and make The micronucleus rate of bone marrow PEC cells was inhibited, which indicated that tea polyphenols had certain protective and therapeutic effects on radiation injury ^[8] .

my country is the birthplace of traditional Chinese medicine, with extremely rich resources. After thousands of years of inheritance, it has a very systematic theory. However, in recent years, the modernization of traditional Chinese medicine has been relatively slow, which has led to restrictions on the development of traditional Chinese medicine. fully accepted. Using microorganisms to ferment traditional Chinese medicine is an important innovation in the history of modernization of traditional Chinese medicine, and it has opened up a new way for the development of traditional Chinese medicine. Modern Chinese medicine fermentation and pharmaceutical technology is based on inheriting the traditional Chinese medicine fermentation and processing method, combined with modern bioengineering technology methods including fermentation technology, etc., to change the performance of Chinese medicine in a targeted manner, or to combine them in a targeted manner according to the characteristics of Chinese medicine. Single strain, or directed fermentation with mixed strains. Various chemical reactions such as oxidation, reduction, esterification, hydroxylation, and methylation of traditional Chinese medicine substrates are carried out by enzymes produced by microorganisms during the fermentation process ^[9] . After fermentation, it can bring many benefits to traditional Chinese medicine, first of all, it will affect the active ingredients in traditional Chinese medicine. During the metabolic process, microorganisms secrete various enzymes such as cellulase, protease, and ligninase, which reduce the mass transfer resistance of active ingredients in traditional Chinese medicine and make them more fully extracted ^[10] . In addition, because microbial fermentation is carried out under normal temperature and pressure, the conditions are mild, which avoids the destruction of the active ingredients of traditional Chinese medicine, and through the action of some special enzymes, some ingredients in traditional Chinese medicine can be modified and transformed to degrade biological macromolecules So as to achieve the purpose of improving the efficacy of traditional Chinese medicine or producing new efficacy ^[11] .

references:

[1] Yang Yingxue, Chen Jianye, Fei Zhonghai, et al. Study on the protective effect of astragalus total flavonoids on radiation-damaged mice [J]. Journal of Chongqing Medical University, 2010, 35(4): 504-507.

[2] Xu Zhengmin, Chen Jianye, Li Xianfu, etc. Anti-radiation damage effect of total flavonoids of astragalus on human normal bone marrow mesenchymal cells [J]. Pharmacology and Clinic of Traditional Chinese Medicine, 2011, 27 (3) 39-41.

[3] Chen Xiwen, Guan Minqiang. Experimental study on the effect of Panax notoginseng saponins against radiation damage in mice [J]. Chinese Journal of Radiation Medicine and Protection, 2005, 25(6): 559-560.

[4] Zou Dan, Qiao Hailing, Quan Hongxun, et al. The antagonism effect of notoginsenosides on radiation-induced bone marrow suppression in mice [J]. Chinese Journal of Radiation Medicine and Protection, 2000, 20(6): 413-415.

[5] Yang Xianqiang, Wang Yuefei, Chen Liuji, etc. Chemistry of Tea Polyphenols [M]. Shanghai: Shanghai Science and Technology Press, 2003, 1-7, 12, 109-125.

[6] Katiyar SK, Elmets CA. Green tea polyphenolic antioxidants and skin photo protection [J]. Int JOncol, 2001, 18(6): 1307～1313.

[7] Wu Liangyu, Lin Jinke. Advances in Research on Anti-radiation of Tea Polyphenols [J]. Tea, 2011, 37(4): 213-217.

[8] Cao Mingfu. Protective effect of tea polyphenols on radiation damage in mice [J]. Tea Science, 1998, 18(2): 139-144.

[9] Qin Chuanguang, Li Shijie, Ding Yan, etc. Application of Fermentation Engineering in Pharmaceutical Research and Production [J]. Journal of Hubei Institute of Technology, 2000, 15(1): 67～70.

[10] Ge Xizhen. The Application of Fermentation in the Research of Traditional Chinese Medicine [J]. Shizhen Guoyi Guoyao, 2008, 19(2): 386-387.

[11] Ruan Xiaodong, Zhang Huiwen, Cai Yinghui, et al. The application of microorganisms in the biotransformation of traditional Chinese medicines [J]. Chinese Herbal Medicine, 2009, 40(1): 149-152.

Invention content:

According to modern scientific and technological research results, the present invention selects Astragalus membranaceus, Panax notoginseng and green tea as experimental raw materials, and its main functional components are flavonoids, saponins and tea polyphenols in sequence. With the purpose of fully leaching the above three active ingredients in the fermentation broth, the best combination of edible fungi for the biotransformation of traditional Chinese medicines was screened and the fermentation medium and fermentation conditions were optimized. The method of orthogonal test was used to formulate the fermentation products, and the physiological and biochemical indexes of the anti-radiation effect of the better formula were determined. The specific technical content is as follows:

1. The traditional Chinese medicine Astragalus was used as the fermentation substrate, and the edible fungus Ganoderma lucidum was used to ferment it, and the change of the total flavonoid content in the fermentation broth was investigated at different times to determine the total flavonoids in the fermentation broth when the edible fungus Ganoderma lucidum fermented Astragalus on the eighth day. The highest content is 2.305mg/g. And through single factor and orthogonal test, with the total flavonoid content in the fermentation liquid as the index, the medium and fermentation conditions of the edible fungus Ganoderma lucidum fermented astragalus were optimized, and the best medium was soluble starch 2g/100mL, potassium nitrate 0.4g/100mL, potassium dihydrogen phosphate 0.1g/100mL, magnesium sulfate 0.1g/100mL, the best fermentation conditions are 10% inoculation amount, 8g/100mL astragalus, temperature 33°C, initial pH value 5.5. Finally, the optimized process was used to verify the total flavonoid content in the fermentation broth was 2.411mg/g.

2 Taking the traditional Chinese medicine Panax notoginseng as the fermentation substrate, using the edible fungus Polyporus chinensis to ferment it, and investigating the change of the total saponin content in the fermentation liquid at different times, it is determined that when the edible medicinal fungus Polyporus polyporus is fermented to the 8th day, The total saponin content in the fermentation broth was the highest, which was 69.063mg/g. And through single factor and orthogonal experiment, with the total saponin content in the fermentation liquid as an index, the culture medium and fermentation conditions of the edible and medicinal fungus Polyporus chinensis fermented with Panax notoginseng were optimized, and the best medium was obtained as soluble starch 3g/100mL, Peptone 0.2g/100mL, Potassium dihydrogen phosphate 0.3g/100mL, Magnesium sulfate 0.1g/100mL, the best fermentation conditions are 10% inoculum amount, 5g/100mL Panax notoginseng, temperature 25℃, initial pH value 7. Finally, the optimized process was used to verify that the total saponin content in the fermentation broth was 72.329mg/g.

3 Using the traditional Chinese medicine green tea as the fermentation substrate, ferment it with the edible fungus Lentinus edodes, and investigate the changes of tea polyphenols content in the fermentation broth at different times, and determine that when the edible medicinal fungus Lentinus edodes is fermented to the sixth day, the fermentation broth The content of polyphenols in Chinese tea is the highest, which is 166.889mg/g. And through single factor and orthogonal experiment, taking the content of tea polyphenols in the fermentation liquid as the index, the culture medium and fermentation conditions of the edible and medicinal fungus Lentinus edodes fermented green tea were optimized, and the best medium was glucose 4g/100mL, yeast extract 0.2g/100mL, potassium dihydrogen phosphate 0.3g/100mL, magnesium sulfate 0.1g/100mL, the best fermentation conditions are 8% inoculum amount, 8g/100mL of traditional Chinese medicine, temperature 25 ℃, initial pH value 7. Finally, the optimized process was used to verify that the content of tea polyphenols in the fermentation broth was 170.749mg/g.

4 Taking the number of white blood cells in the peripheral blood on the 3rd and 10th day after irradiation as the evaluation index, adopt the orthogonal design method to formulate the dosage ratio formula of the fermented products of Astragalus membranaceus, Panax notoginseng and green tea, and select the best formula as the formula 7, that is, Panax notoginseng fermentation broth 0.4g/kg d, astragalus fermentation broth 3.8g/kg d, green tea fermentation broth 2.25g/kg d; recipe 2, Panax notoginseng fermentation broth 0.8g/kg d, Astragalus fermented liquid 2.5g/kg·d, green tea fermented liquid 2.25g/kg·d.

5 pairs of irradiated mice peripheral blood white blood cell (WBC) count, immune organ index, endogenous splenic colony forming unit (CFU-S), bone marrow nucleated cell (BMC) count, bone marrow polychromatic erythrocyte micronucleus rate (PCE ) five indicators were measured, and the results showed that both prescription 2 and prescription 7 had a certain anti-radiation effect.

At present, the research on anti-radiation effect of traditional Chinese medicine is increasing day by day, but the effect is significantly developed less. The present invention adopts the method of microbial fermentation, the purpose is to improve drug efficacy, obtain a dose of prescription with good anti-radiation effect, open up a new idea for the research of anti-radiation products with traditional Chinese medicine as raw materials, and at the same time in Astragalus, Panax Further applied research on green tea provides a theoretical basis.

Detailed ways:

Embodiment one: the preparation of astragalus fermented liquid

The traditional Chinese medicine Astragalus is used as the fermentation substrate, and the edible fungus Ganoderma lucidum is used to ferment it. The fermentation medium is soluble starch 2g/100mL, potassium nitrate 0.4g/100mL, potassium dihydrogen phosphate 0.1g/100mL, magnesium sulfate 0.1g/ 100mL, the best fermentation conditions are 10% inoculum size, 8g/100mL astragalus addition, 33°C temperature, 5.5 initial pH value, and 8-10d culture time to obtain astragalus fermentation broth, which is centrifuged at 3500r/min for 15min, and the supernatant The liquid can be directly used as food, health products and medicines, or used as raw materials for preparing food, health products and medicines.

Embodiment two: the preparation of notoginseng fermented liquid

The traditional Chinese medicine Astragalus is used as the fermentation substrate, and the edible fungus Polyporus chinensis is used to ferment it. The fermentation medium is 3g/100mL of soluble starch, 0.2g/100mL of peptone, 0.3g/100mL of potassium dihydrogen phosphate, and 0.1g/100mL of magnesium sulfate. 100mL, the optimal fermentation conditions are as follows: 10% inoculation amount, 5g/100mL Panax notoginseng addition, 25°C temperature, 7 initial pH value, 8-10d cultivation time to obtain Panax notoginseng fermentation broth, centrifuged at 3500r/min for 15min, the The supernatant can be directly used as food, health products and medicines, or used as raw materials for preparing food, health products and medicines.

Embodiment three: the preparation of green tea fermented liquid

Using traditional Chinese medicine green tea as the fermentation substrate, it is fermented with the edible fungus Lentinus edodes. The fermentation medium is glucose 4g/100mL, yeast extract 0.2g/100mL, potassium dihydrogen phosphate 0.3g/100mL, magnesium sulfate 0.1g/100mL , the optimal fermentation conditions are 8% inoculum amount, 8g/100mL of traditional Chinese medicine, 25°C temperature, 7 initial pH value, and 6-8d cultivation time to obtain green tea fermentation liquid, which is centrifuged at 3500r/min for 15min, and the supernatant It can be directly used as food, health products and medicines, or used as raw materials for preparing food, health products and medicines.

Example 4: Orthogonal design method to optimize the anti-radiation formula of traditional Chinese medicine fermented products

Experimental animals: healthy female Kunming mice, 18-22g, clean grade.

Irradiation conditions: The synergy medical linear accelerator was used to irradiate the whole body of the mice once, the irradiation dose was 3Gy, the dose rate was 0.5Gy/min, the irradiation time was 6min, and the distance between the bullseye was 100cm.

Orthogonal design grouping: use L ₉ (3 ³ ) three-factor three-level orthogonal table to carry out test grouping, the factor level table is shown in Table 1, in which factor A is astragalus fermented liquid, factor B is notoginseng fermented liquid, factor C is green tea For the fermented liquid, the level dosage of each factor is represented by the amount of original raw medicinal material contained in the fermented liquid fed per kilogram of mice every day. Another group of irradiation model control group was set up, and each test group was fed continuously for 20 days before irradiation, once a day. Continue to feed after irradiation.

Table 1 Level table of test grouping factors

Table 1 The table of the grouping factor of experiment

Peripheral blood leukocyte count: before irradiation, on the 3rd day after irradiation and on the 10th day after irradiation, cut the mouse tail vein, take 20 μL of blood, add it to 0.38mL of 1% hydrochloric acid solution, mix well, add Go to the hemocytometer, and count the total number of white blood cells in the four large squares of the counting pool under the microscope.

Experimental results:

(1) Orthogonal test results on the 3rd day after irradiation: the effects of the three fermentation broths obtained by orthogonal design on the peripheral blood leukocyte count of mice (3 days before irradiation and 3 days after irradiation) are shown in Table 2 , It can be seen from the table that there is no significant difference in the number of WBCs in each experimental group 3 days before irradiation, and follow-up experiments can be carried out. 3 days after irradiation, the WBC counts in each experimental group were significantly lower than those 3 days before irradiation, indicating that the model of radiation damage to mice was successfully established. The results of the orthogonal test show that the degree of influence of the three factors on the WBC of mice in this experiment is C>B>A, that is, green tea fermented liquid> astragalus fermented liquid> Panax notoginseng fermented liquid; the best fermented liquid formula is A ₁ B ₃ C ₂ , that is, prescription 7, in which Sanqi fermentation liquid is 0.4g/kg·d, astragalus fermentation liquid is 3.8g/kg·d, and green tea fermentation liquid is 2.25g/kg·d. And the WBC of group 7 of the prescription was 5.71×10 ⁹ /L three days after irradiation, significantly higher than that of the model group of 4.81×10 ⁹ /L, so it was determined that the experimental results of group 7 of the prescription were positive.

Table 2 L ₉ (3 ³ ) orthogonal test results on the third day after irradiation

Table 2 The L ₉ (3 ³ )orthogonal test results of the third day of postirradiation

(2) Orthogonal test results on the 10th day after irradiation: Table 3 shows the results of the effects of the three fermentation broths obtained by orthogonal design on the number of peripheral blood leukocytes in mice 3 days before irradiation and 10 days after irradiation. There was no significant difference in the number of WBC in each experimental group in the first 3 days, and follow-up tests could be carried out. The number of WBC in each experimental group 10 days after irradiation was significantly lower than that 3 days before irradiation, indicating that the model of radiation damage to mice was successfully established. It can be seen from Table 3 that the degree of influence of the three factors on WBC of mice in this experiment is C>A>B, that is, green tea fermentation liquid> Sanqi fermentation liquid> Astragalus membranaceus fermentation liquid; the best fermentation liquid formula is A ₂ B ₂ C ₂ , that is, prescription 2, in which the fermented liquid of notoginseng is 0.8g/kg·d, the fermented liquid of astragalus is 2.5g/kg·d, and the fermented liquid of green tea is 2.25g/kg·d. And the WBC of the prescription group 2 was 7.06×10 ⁹ /L 10 days after irradiation, which was significantly higher than the 6.26×10 ⁹ /L of the model group, so it was judged that the experimental result of the prescription group 2 was positive.

Table 3 Orthogonal test results of L ₉ (3 ³ ) on the 10th day after irradiation

Table 3 The L ₉ (3 ³ )orthogonal test results of the tenth day of postirradiation

Embodiment 5: Determination of physical and chemical indicators of better anti-radiation prescription

Experimental animals: healthy female Kunming mice, 18-22g, clean grade.

Irradiation conditions: The synergy medical linear accelerator was used to irradiate the whole body of the mice once, the irradiation dose was 3Gy, the dose rate was 0.5Gy/min, the irradiation time was 6min, and the distance between the bullseye was 100cm.

Experimental drugs: Fermented liquid recipe 2 and fermented liquid recipe 7 were prepared. See Table 4 for the dosage (the amount of raw medicinal material contained in the fermented liquid fed per kilogram of mice per day). In addition, decoction prescription 2 and decoction prescription 7 were prepared according to the dosage of the fermentation broth prescription.

Table 4 Fermentation broth prescription dose setting table

Table 4 The installatio table of fermented liquid group prescriptions dose

Animal grouping: 140 mice were randomly divided into 7 groups after observation for one week, namely, blank control group, radiation model control group, positive control group, fermentation liquid group 2, fermentation liquid group 7, and water decoction group 2 7 groups, 20 rats in each group. Each test group was given continuous gavage for 20 days before irradiation, once a day. The positive control group was given compound donkey-hide gelatin syrup 8.5mL/kg·d, the blank control group and the model control group were given 0.5ml distilled water, all groups were given one-time whole-body irradiation under the same irradiation conditions, and continued to be fed after irradiation. The mice were sacrificed on the 3rd day and the 10th day for the determination of physical and chemical indicators.

Animal experiments and experimental results:

(1) Effects of traditional Chinese medicine prescriptions on the number of peripheral blood leukocytes in irradiated mice: before irradiation, on the 3rd day after irradiation and on the 10th day after irradiation, the mouse tail vein was cut, 20 μL of blood was taken, and added to 0.38 mL 1% hydrochloric acid solution, mixed thoroughly, added to a hemocytometer, and counted the total number of white blood cells in the four large squares of the counting pool under a microscope. The results are shown in Table 5.

Table 5 Effects of traditional Chinese medicine prescriptions on the number of peripheral blood leukocytes in irradiated mice

Table 5 The effect of the traditional chinese group prescriprions to peripheral blood leucocyte; peripheral white blood cell of mice by radiated

Note: Compared with the blank group, ^* p<001; compared with the model group, ^a p<001, ^b p<005

It can be seen from Table 5 that the number of peripheral blood white blood cells (WBC) in each experimental group did not change significantly in mice 3 days before irradiation, and there was no statistical significance. On the 3rd day after irradiation, the WBC counts in each administration group and model group were significantly different from those in the blank control group (p<0.01); compared with the model group, the WBC counts in each administration group were increased, and the difference in the positive group was extremely significant (p<0.01), and the other groups were significantly different (p<0.05).

On the 10th day after irradiation, the differences in WBC counts between each administration group and the model group were extremely significant compared with the blank control group (p<0.01); compared with the model group, the WBC counts in the fermentation liquid group 2 and the positive group were extremely significant increased (p<0.01), and the fermented liquid prescription 7 group and the water decoction prescription 2 and prescription 7 groups had a significant increase (p<0.05).

(2) The effect of traditional Chinese medicine prescriptions on the immune organ index of irradiated mice: the mice were killed by cervical dislocation, and the weight of the mice was accurately weighed by an electronic balance. The mice were dissected, and the liver, spleen and thymus were weighed to calculate the immune organs index. The results are shown in Table 6.

Table 6 Effects of traditional Chinese medicine prescriptions on immune organ indexes of irradiated mice

Table 6 The effect of the traditional chinese group prescriprions to immune organ Exponent of mice

Note: Compared with the model group, ap<001, bp<005

It can be seen from Table 6 that after the mice were irradiated for 10 days, there was no significant change in the body weight of each experimental group, which was not statistically significant. Compared with the blank control group, there was no significant difference in liver index, spleen index and thymus index in each experimental group (p>0.05). Compared with the model group, the liver index of the 7th group of the fermented liquid prescription and the positive group had a very significant increase (p<0.01), the 2nd group of the fermented liquid recipe significantly improved (p<0.05), and the rest of the experimental groups did not increase significantly (p> 0.05); the spleen index of fermentation liquid prescription group 7 and the positive group had a significant increase (p<0.05), although the remaining experimental groups had a rising trend, it was not significant (p>0.05); fermentation liquid prescription 2, prescription The thymus index of group 7 and decoction group 7 and the positive group all increased significantly (p<0.05), but the decoction group 2 did not increase significantly (p>0.05).

(3) The influence of traditional Chinese medicine prescriptions on the endogenous spleen colony forming units of irradiated mice: After the mice were sacrificed, the fresh spleen was soaked in Bouin's solution for 24 hours, and decolorized with 70% ethanol solution after removal , After removing most of the yellow color, observe the spleen colony forming units under a magnifying glass, that is, the number of spleen nodules. The results are shown in Table 7.

Table 7 Effects of traditional Chinese medicine prescriptions on endogenous spleen colony forming units in irradiated mice

Table 7 The effect of the traditional chinese group prescriprions to endogenous spleen colony fomingunit-spleen of mice

Note: Compared with the blank group, ^* p<001; compared with the model group, ^b p<005

It can be seen from Table 7 that on the 3rd day and the 10th day after irradiation, the number of endogenous splenic colony-forming units in mice in each experimental group, that is, the number of spleen nodules, was increased compared with the blank control group, and the difference was extremely significant (p <0.01). Compared with the model group, on the 3rd day after irradiation, the number of spleen nodules in each treatment group increased, but the difference was not significant (p>0.05); Compared with the model group, the number of knots was significantly increased (p<0.05). Although there was no significant difference in the other administration groups, it can be seen from the data that the fermented liquid prescription 7 groups had a significant upward trend.

(4) The effect of traditional Chinese medicine formula on the number of nucleated cells in the bone marrow of irradiated mice: Take one side of the femur of the mouse, remove the muscle tissue, draw 1 mL of Hank's solution with a disposable sterile syringe, and wash the bone marrow cells in the femur repeatedly , until all the bone marrow cells were washed out, placed in a 1.5mL centrifuge tube, and finally filtered all the bone marrow cell liquid with a No. 4 needle to make it fully and evenly dispersed, and counted under a microscope. The results are shown in Table 8.

Table 8 Effects of traditional Chinese medicine prescriptions on the number of nucleated cells in the bone marrow of irradiated mice

Table 8 The effect of the traditional chinese group prescriprions to marrow karyocyte count of mice

Note: Compared with the blank group, ^* p<001: compared with the model group, ^a p<001, ^b p<005

It can be seen from Table 8 that the number of bone marrow nucleated cells (BMC) in the mice was significantly lower than that of the blank control group on the 3rd day and the 10th day after irradiation (p<0.01), indicating that radiation has a significant effect on the number of bone marrow nucleated cells in mice. Play damage. On the 3rd day after irradiation, the number of BMCs in the positive group and the fermented liquid formula 2 and 7 groups was significantly higher than that of the model group (p<0.05); on the 10th day after irradiation, the BMC counts of the fermented liquid formula 7 groups were significantly higher than those in the model group. The model group had a very significant increase (p<0.01), the fermented liquid prescription group 2, the water decoction prescription group 2 and the positive group had a significant increase (p<0.05).

(5) The effect of traditional Chinese medicine prescription on the micronucleus rate of bone marrow polychromatic erythrocytes in irradiated mice: take the other femur of the mouse, remove the muscle tissue, subtract the epiphyses at both ends with sharp scissors, and draw a certain volume of For calf serum, the bone marrow cells were washed out and placed on slides, routinely smeared, dried at room temperature, fixed in methanol solution for 10 minutes, taken out, and then stained in Giemsa staining solution for 10-15 minutes. Rinse with distilled water immediately after staining, and after natural drying, observe the number of micronucleated cells per thousand bone marrow polychromatic erythrocytes under an oil microscope, that is, the micronucleus rate of bone marrow polychromatic erythrocytes. The results are shown in Table 9.

Table 9 Effects of traditional Chinese medicine prescriptions on the micronucleus rate of bone marrow polychromatic erythrocytes in irradiated mice

Table 9 The effect of the traditional chinese group prescriprions to Bone marrow polychromatic erythrocytemicronuclear rates of mice

Note: Compared with the blank group, ^* p<001: compared with the model group, ^a p<001, ^b p<005

It can be seen from Table 9 that the polychromatic erythrocyte micronucleus rate (PCE) in the bone marrow of mice in each experimental group increased on the 3rd day and the 10th day after irradiation compared with the blank control group, and the difference was extremely significant (p<0.01), indicating that the mouse radiation injury model was established successfully. On the 3rd day after irradiation, compared with the model group, the PCE of the fermented liquid prescription 2 group and the positive group had a significant decrease (p<0.05). Although the other administration groups had a downward trend, the difference was not significant (p>0.05 ); on the 10th day after irradiation, compared with the model group, the PCE of the positive group was significantly reduced (p<0.01), and the fermentation broth group 2 and group 7 were significantly reduced (p<0.05), while the water Although the decoction group 2 and group 7 decreased, it was not significant (p>0.05).

Claims (6)

1. A Radix Astragali fermented liquid is characterized in that taking the traditional Chinese medicine Astragalus Radix Astragali as a fermentation substrate, utilizing the edible fungus Ganoderma lucidum to ferment it, adopting fermentation medium as soluble starch 2g/100mL, potassium nitrate 0.4g/100mL, dihydrogen phosphate Potassium 0.1g/100mL, magnesium sulfate 0.1g/100mL, the best fermentation conditions are inoculum size 10%, astragalus addition 8g/100mL, temperature 33°C, initial pH value 5.5, culture time 8-10d, to obtain astragalus fermentation broth After being centrifuged at 3500r/min for 15 minutes, the supernatant can be directly used as food, health products and medicines, or used as raw materials for preparing food, health products and medicines. 2. A Radix Notoginseng fermented liquid is characterized in that taking the traditional Chinese medicine Radix Astragali as the fermentation substrate, utilizing the food medicine fungus Polyporus chinensis to ferment it, adopting the fermentation medium as soluble starch 3g/100mL, peptone 0.2g/100mL, diphosphate Potassium hydrogen 0.3g/100mL, magnesium sulfate 0.1g/100mL, the best fermentation conditions are 10% inoculum amount, 5g/100mL Panax notoginseng, temperature 25°C, initial pH value 7, culture time 8-10d, and three 7. Fermentation liquid, centrifuged at 3500r/min for 15min, the supernatant can be directly used as food, health products and treats, or used as raw materials for preparing food, health products and medicines. 3. A green tea fermented liquid is characterized in that it is fermented substrate with Chinese medicine green tea, and it is fermented by using edible fungus shiitake mushrooms, adopting fermentation medium as glucose 4g/100mL, yeast extract 0.2g/100mL, potassium dihydrogen phosphate 0.3g/100mL, magnesium sulfate 0.1g/100mL, the optimal fermentation conditions are 8% inoculum amount, 8g/100mL of traditional Chinese medicine, temperature 25°C, initial pH value 7, incubation time 6-8d, to obtain green tea fermentation liquid, After being centrifuged at 3500r/min for 15 minutes, the supernatant can be directly used as food, health products and medicines, or used as raw materials for preparing food, health products and medicines. 4. a Chinese medicine fermented liquid, it is characterized in that three kinds of Chinese medicine fermented liquids prepared by the method described in claims 1 to 3 are carried out dose proportioning prescription, this prescription has certain anti-radiation effect, can be used in the preparation of anti-radiation medicine, Applied in food and health products. 5. The traditional Chinese medicine fermented liquid according to claim 4, characterized in that the fermented liquid can be prepared into soft capsules, hard capsules, dripping pills, tablets, dispersible capsules alone or in combination with other medicines or by adding pharmaceutically acceptable adjuvants. Tablets, granules, injections, oral solutions, suppositories, orally, skin, mucous membrane, rectal application. 6. the present invention is to adopt the method for shaking flask to carry out, as carrying out enlarged production, can adopt modes such as various bioreactors, fermentors to carry out fermentation treatment according to the basic principle of the present invention, except the edible fungus Ganoderma ganoderma used in the present invention in addition , Polyporus, Lentinus edodes, but also other large edible and medicinal fungi such as oyster mushroom, black fungus, Flammulina velutipes etc., this is what researchers in this field can understand.