CN101537128B - Application and products of dendrobium nobile total alkaloids in the preparation of medicines for treating senile dementia - Google Patents

Abstract

The invention discloses the application of total alkaloids of Dendrobium nobile in the preparation of medicines for treating Alzheimer's disease and the product thereof, wherein the total alkaloids of Dendrobium nobile are lipophilic total alkaloid components extracted from Dendrobium nobile. Compared with the prior art, the invention clarifies the pharmacological effect and mechanism of total alkaloids of Dendrobium nobile in treating Alzheimer's disease by studying the protective effect of total alkaloids of Dendrobium nobile on nerve cells and cerebral ischemia models and the influence on the learning and memory ability of dementia rats, and provides a pharmacological basis for the development and application of medicinal materials of Dendrobium nobile.

Description

Application and products of dendrobium nobile total alkaloids in the preparation of medicines for treating senile dementia

Technical field: The present invention relates to the application and products of Dendrobium nobile total alkaloids in the preparation of medicines for treating senile dementia, belonging to the technical field of traditional Chinese medicine.

Background technology: Alzheimer's disease (AD) is a disease related to aging, and is the main cause of dementia in the elderly over 65 years old. With the emergence of my country's population aging problem, AD has become an important issue of family burden and social medical insurance. The clinical manifestations of AD are progressive mental decline, mental and motor disorders, and the main pathological features are senile plaques (SP), neurofibrillary tangles (NFT), vascular amyloidosis, neuronal degeneration and loss. AD is a heterogeneous disease with multiple causes, and the exact etiology and pathogenesis are still unclear. Existing studies have shown that its pathogenesis may involve gene defects, central neurotransmitter disorders, metabolic disorders, oxidative stress, neuronal apoptosis, environmental toxins, and chronic inflammation. At present, there is no ideal treatment method (or drug) for senile dementia at home and abroad. On the one hand, we continue to study the pathogenesis of AD in depth; Inflammatory drugs, cholinesterase inhibitors, calcium blockers, excitatory amino acid antagonists, etc. are used for comprehensive treatment to relieve symptoms or delay the course of the disease. In pharmacology, on the one hand, identifying drug targets through genomics and proteomics studies of diseases is an ideal way to find therapeutic drugs for related diseases, for example, the abnormality of β-amyloid precursor protein (APP) degrading enzymes Expression, M receptor protein expression, NDMA receptor expression, etc. provide directions for determining the target of drug action; Drugs for the treatment of AD are of great significance.

Dendrobium (Dendrobium) is a precious Chinese medicinal material in traditional Chinese medicine, and its main chemical components are polysaccharides, alkaloids, amino acids, phenols, and volatile oils. Dendrobium alkaloids can cause hyperpolarization, and as a possible antagonist of β-alanine or taurine, have antitumor activity and immunomodulatory activity. Studies have shown that Dendrobium extract can significantly inhibit the secretion of IL-10, TNF-α, iNOS and IL-8 from mouse peritoneal macrophages induced by bacterial lipopolysaccharide (LPS), but its mechanism of action is still unclear and needs further investigation Research. Dendrobium nobile is a kind of Dendrobium nobile. Dendrobium nobile extract can antagonize atropine and promote intestinal peristalsis; reduce blood viscosity, antagonize arachidonic acid and collagen-induced platelet aggregation in rabbits; have anti-free radical damage; simulate monoamine oxidase Inhibitors that modulate monoamine transmitters. At present, there is no research report on the effect of Dendrobium nobile extract on senile dementia.

Invention content:

The object of the present invention is to provide the application and products of Dendrobium nobile total alkaloids in the preparation of medicines for treating senile dementia. The present invention clarifies the pharmacological action mechanism of Dendrobium nobile total alkaloids in treating senile dementia by studying the protective effect of Dendrobium nobile total alkaloids on nerve cells and cerebral ischemia models and the learning and memory ability of dementia rats. The further development and application of Dendrobium officinalis provides a pharmacological basis.

The present invention is constituted as follows: the application of total alkaloids of Dendrobium nobile in the preparation of medicine for treating senile dementia.

The total alkaloids of Dendrobium nobile are lipophilic total alkaloid components extracted from Dendrobium nobile.

The preparation method of the aforementioned total alkaloids of Dendrobium nobile is as follows: take the medicinal material of Dendrobium nobile, grind it into a coarse powder, add ethanol to extract 3 times, recover the ethanol until it has no alcohol smell, adjust the pH to 3-4 with an acid solution, filter, and the filtrate Adjust the pH to 10 with alkaline solution, then extract with appropriate amount of chloroform for 5 to 8 times, collect the chloroform layer, recover the chloroform, and dry to obtain the product.

The preferred preparation method is as follows: take the medicinal material of Dendrobium nobile, crush it into a coarse powder, add 95% ethanol to reflux and extract for 3 times, recover the ethanol until it has no alcohol smell, adjust the pH to 3-4 with hydrochloric acid, filter, and filter the filtrate with 10% Adjust the pH to 10 with ammonia water, then extract with appropriate amount of chloroform for 5 to 8 times, collect the chloroform layer, recover the chloroform, and dry to obtain the product.

The content of lipophilic total alkaloids in the aforementioned Dendrobium nobile total alkaloids is more than or equal to 60%.

A pharmaceutical preparation for treating senile dementia is an oral preparation or an injection prepared by taking total alkaloids of Dendrobium nobile as a raw material and adding appropriate amount of auxiliary materials.

The pharmaceutical preparation is tablet, capsule, granule, soft capsule, drop pill, syrup, sublingual tablet or injection.

Dendrobium nobile has the effects of promoting body fluid and benefiting stomach, clearing away heat and nourishing yin. The applicant has conducted in-depth and meticulous research on its chemical composition, pharmacological effects, and mechanism of action, and found that the alkaloid components contained in the medicinal material of Dendrobium nobile can significantly Reduce the injury of primary cultured neurons caused by hypoxia and glucose deficiency/reoxygenation and reglucose, and have a protective effect on the acute cerebral ischemia model in rats. It can alleviate the inflammation of rat nerve cells induced by LPS and improve its learning and memory ability, and can improve LPS. Induced learning and memory impairment in rats, alleviated neuronal apoptosis. The main content and results of the experimental study are as follows:

1. Experimental content

1. The primary nerve cells of cultured rats were subjected to hypoxia and glucose deficiency injury, and the protective effect of total alkaloids of Dendrobium nobile on neuron cells was observed:

1) The protective effect of Dendrobium nobile total alkaloids on nerve cell damage caused by hypoxia and glucose deficiency was observed by MTT method;

2) Observing the effect of total alkaloids of Dendrobium nobile on the apoptosis of neuron cells from the cell morphology by fluorescence staining and flow cytometry;

3) The effect of sugar deficiency/reoxygenation and re-sugar on the mitochondrial membrane potential (MMP) of nerve cells and the effect of total alkaloids in Dendrobium nobile;

4) The effect of hypoxia and sugar deficiency/reoxygenated sugar on free Ca ²⁺ in nerve cells and the effect of total alkaloids in Dendrobium nobile;

5) RT-PCR was used to observe the effect of the total alkaloids of Dendrobium nobile on the apoptosis genes caspase-3 and caspase-12 of hypoxia-deficient/reoxygenation and reglucose neuronal cells.

2. The focal cerebral ischemia model of right middle cerebral artery occlusion in rats was made by thread-tethering method, and the protective effect of total alkaloids of Dendrobium nobile on its injury was observed:

1) Measure cerebral infarct volume, ischemic side cortex and hippocampus for HE staining, and observe pathological changes;

2) The content of MDA and NO, the activity of SOD and NOS and the change of MDA and SOD were determined by tissue homogenate;

3) Using the Real time RT-RCR method, observe the expression of caspase-3 and caspase-8 mRNA in brain tissue.

3. Injection of lipopolysaccharide (LPS) into the rat model of senile dementia, observe the protective effect of total alkaloids of Dendrobium nobile on LPS-induced learning and memory dysfunction in rats, and explore its mechanism:

1) The Morris water maze was used to test the spatial discrimination learning and memory ability of rats;

2) Immunohistochemical method to detect the expression of MHC-II (OX-6), TNF-α, IL-1β, S-100β, iNOS and COX-2 in the cerebral cortex and hippocampus; immunohistochemical method to detect the expression of Aβ in the hippocampus _1-42 fragment content, and semi-quantitative analysis through BI2000 image analysis system;

3) Real time RT-PCR to detect the mRNA expression of APP ₆₉₅ , BACE1, PS-1, caspase-3, caspase-8, TNFα, IL-1β, iNOS, NF-κB, and COX-2 in the hippocampus;

4) Observe the morphological changes of neurons under the HE staining light microscope, and observe the ultrastructural changes of the hippocampal neurons under the electron microscope.

4. Statistical processing

The results were expressed as x±s, and were analyzed using SPSS12.0 statistical software, and the comparison between groups was performed with one-way analysis of variance combined with LSD test.

2. Experimental results

1. Effect of total alkaloids (DNLA) of Dendrobium nobile on the damage of primary cultured nerve cells in rats induced by hypoxia and glucose deficiency

1) Compared with the blank group, the absorbance value of MTT decreased significantly after hypoxic injury: there were significant differences between the absorbance values of each model group and the blank group at the same time; The dose group can increase the MTT absorbance value, which is different from that of the model group at the same time point (P<0.05); the hypoxia 2h/reoxygenation 3h dosage groups, the hypoxia 2h/reoxygenation 12h middle and high dose groups, and the hypoxia 2h /Reoxygenation 24h high-dose group can increase the MTT absorbance value, which is different from that of the model group at the same time point.

2) The leakage rate of LDH increased significantly after hypoxic injury: there were significant differences between each model group and the blank group at the same time, the middle and high dose groups of hypoxia 1h, each dose group of hypoxia 2h, and the high dose group of hypoxia 4h It can reduce the leakage rate of LDH; the hypoxia/reoxygenation 3h medium and high dose group, the hypoxia 2h/reoxygenation 12h medium and high dose group, the hypoxia 2h/reoxygenation 24h high dose group can reduce the LDH leakage rate. There were differences among the model groups.

3) The apoptosis rate of nerve cells increased significantly after hypoxia and glucose deficiency for 2h/reoxygenation and reglucose for 12h: there was a significant difference between the model group and the blank group; each dose group of total alkaloids of Dendrobium nobile could reduce cell apoptosis The rate was different from that of the model group.

4) After hypoxia and glucose deficiency for 2h/reoxygenation and reglucose for 12h after injury, the intracellular free calcium concentration was significantly increased: the model group had a significant difference compared with the blank group; each dose group could reduce the intracellular free Ca2 ⁺ concentration, compared with There were significant differences among the model groups.

5) After hypoxia and glucose deficiency for 2h/reoxygenation and reglucose for 12h, MMP decreased significantly, and there was a significant difference between the model group and the blank group; each dose group of Dendrobium nobile total alkaloids increased MMP, compared with the model group. Significant difference.

6) The results of Real time RT-RCR test showed that the expressions of casPase-3 and casPase-12 were significantly increased after 12 hours of hypoxia-deficiency-deficiency-glucose-2 quasi-reoxygenation and re-glucose injury, and there was a significant difference between the cells in the model group and the blank group; Dendrobium alkaloids of each dose group can reduce the expression of caspase-3 and caspase-12, compared with the model group, there is a significant difference.

7) Morphological observation: After hypoxic injury, the cells lost their refraction, the outline of the cell body was blurred, and the protrusions were reduced or retracted and broken. HE detection found that the cells in the model group also had nuclear pyknosis and dense staining, and the mitochondrial swelling was observed by the electron microscope microstructure. , clear fracture, fuzzy; Dendrobium nobile total alkaloids group damage significantly less than the model group.

2. Protective effect of Dendrobium nobile total alkaloids (DNLA) on cerebral ischemia model of middle cerebral artery occlusion

1) Compared with the model group, the neurological symptom scores of the DNLA high-dose group and the nimodipine group were reduced (P<0.05); the cerebral infarct volumes of the DNLA middle- and high-dose groups and the nimodipine group were reduced (P<0.05); Morphological observation: HE staining revealed that the temporal lobe cortex cells in the model group were edema, the intercellular space was widened, and the cell structure was damaged; the arrangement of hippocampal granule cells was loose and disordered, and the nuclei were condensed. The cell damage in the high-dose group and the nimodipine group was significantly less than that in the model group.

2) Compared with the model group, the contents of MDA and NO in the middle and high dose groups and the nimodipine group decreased, the activity of NOS decreased, the activity of SOD increased (P<0.05), and the ratio of MDA to SOD decreased (P<0.05). 0.05); compared with the model group, the middle and high dose groups and the nimodipine group could down-regulate the expression of caspase-3 and caspase-8 mRNA to varying degrees (P<0.05).

3. Protective effect on lipopolysaccharide-induced senile dementia in rats

1) The results of behavioral tests showed that after 6 days and 13 days of injection of lipopolysaccharide solution into the contralateral ventricle of DNLA, compared with the model control group, DNLA (20mgkg ^-1 d ^-1 , 40mgkg ^-1 d ^-1 , 80mgkg ^-1 d ^-1 ) for 7 days and 14 days can shorten the escape latency and search distance of rats in the positioning navigation experiment, and increase the search time of rats in the quadrant where the original safety island is located and the proportion of swimming distance in the quadrant where the original safety island is located in the space exploration experiment. Distance percentage (P<0.05). The escape latency and search distance of the model rats were significantly longer than those of the control group (P<0.01), suggesting that learning and memory were impaired; both DNLA and ibuprofen could shorten the escape latency and search distance of the model rats.

2) The results of immunohistochemistry showed that it can reduce the optical density values of OX-6, TNF-α, IL-1β, S-100β, iNOS, COX-2 and TNF-α, IL-1β, NF- The expression levels of κB, iNOS and COX-2mRNA; and it was dose-dependent and time-dependent.

3) Seven days and 14 days after LPS modeling, the number of neurons in the hippocampus of the model rats was significantly reduced under the photoelectric microscope, and apoptosis and necrosis of neurons appeared, and the apoptosis and necrosis of neurons were alleviated.

4) Real time RT-PCR test results showed that compared with the blank group, the content of Aβ _1-42 fragments in the hippocampus and the expression of APP695, BACE1, PS-1, caspase-3, and caspase-8 mRNA in the model group increased significantly; Compared with the model group, DNLA (40mg·kg ^-1 ·d ^-1 , 80mg·kg ^-1 ·d ^-1 ) could reduce the contents of Aβ _1-42 fragments and APP ₆₉₅ , BACE1, PS-1, caspase- 3. The expression of caspase-8mRNA, in which the DNLA (40mg·kg ^-1 ·d ^-1 , 80mg·kg ^-1 ·d ^-1 ) group had no significant difference compared with the ibuprofen group, and there was a certain dose dependent and time dependent.

3. Conclusion

1. Dendrobium nobile total alkaloids can significantly reduce the damage of primary cultured neurons caused by hypoxia and glucose deficiency/reoxygenation and reglucose, and its mechanism may involve improving energy metabolism, reducing apoptosis, inhibiting intracellular ca ²⁺ overload and inhibiting Apoptosis-specific proteases and other factors.

2. The total alkaloids of Dendrobium nobile have protective effects on the acute cerebral ischemia model in rats, and the mechanism may involve reducing the generation of oxygen free radicals, anti-oxidative stress, reducing the transcription level of apoptosis genes in the rat brain, and inhibiting nerve cells. apoptosis.

3. The total alkaloids of Dendrobium nobile can relieve LPS-induced nerve cell inflammation in rats and improve their learning and memory ability. 2) reduce the production of NO by inhibiting the expression of iNOS, so as to reduce the damage of NO to nerve cells; 3) protect nerve cells by inhibiting COX-2; 4) inhibit the activation of NF-κB Blocking NF-κB-mediated signaling pathways, thereby attenuating acute brain injury caused by iNOS, COX-2 and other inducible genes.

4. Total alkaloids of Dendrobium nobile can improve LPS-induced learning and memory impairment in rats, and reduce neuronal apoptosis. The mechanism may be related to the inhibition of APP ₆₉₅ , BACE1, PS-1, caspase-3 and caspase-8 mRNA It is related to the decrease of Aβ _1-42 fragment content in the hippocampus.

Compared with the prior art, the present invention clarifies its pharmacological action and role in treating senile dementia by studying the protective effect of total alkaloids of Dendrobium nobile on nerve cells and cerebral ischemia models and on the learning and memory ability of dementia rats. The mechanism provides a pharmacological basis for the development and application of Dendrobium nobile medicinal materials.

Detailed ways:

Embodiment 1 of the present invention: the extraction of total alkaloids of Dendrobium nobile:

Take the medicinal material of Dendrobium nobile, crush it into coarse powder, add 95% ethanol to reflux and extract 3 times, recover the ethanol until it has no alcohol smell, adjust the pH to 3-4 with hydrochloric acid, filter, and adjust the pH of the filtrate to 10 with 10% ammonia water, Then extract with appropriate amount of chloroform for 7-8 times, collect the chloroform layer, recycle the chloroform, and dry to obtain the total alkaloids of Dendrobium nobile; wherein the content of lipophilic total alkaloids is 68.3%.

Embodiment 2 of the present invention: the extraction of total alkaloids of Dendrobium nobile:

Take the medicinal material of Dendrobium nobile, crush it into coarse powder, add ethanol to extract 3 times, recover the ethanol until it has no alcohol smell, adjust the pH to 3-4 with hydrochloric acid, filter, adjust the pH of the filtrate to 10 with sodium hydroxide solution, and then use an appropriate amount of Extract 5-6 times with chloroform, collect the chloroform layer, recycle the chloroform, and dry to obtain the total alkaloids of Dendrobium nobile, wherein the content of lipophilic total alkaloids is 61.9%.

Embodiment 3 of the present invention: the application of total alkaloids of Dendrobium nobile:

Get 50-2000 mg of Dendrobium nobile total alkaloid extract, 0.1-0.40 mg of magnesium stearate, 4-12 mg of sodium carboxymethyl starch, and 50-100 mg of microcrystalline cellulose obtained in Example 1, and extract The product is mixed with sodium carboxymethyl starch and microcrystalline cellulose, sieved, mixed evenly, added an appropriate amount of water or ethanol to granulate, after drying, granulated, added magnesium stearate, and then pressed the granules with a punching device into Tablets, that is, tablets for treating senile dementia. The tablet is taken orally, 3 times a day, 1-3 tablets each time.

Embodiment 4 of the present invention: the application of total alkaloids of Dendrobium nobile:

Get 50-2000 mg of Dendrobium nobile total alkaloid extract, 0.1-0.30 mg of magnesium stearate, 4-12 mg of sodium carboxymethyl starch, and 50-100 mg of starch obtained in Example 2. Mix the sodium methyl starch and starch evenly, add appropriate amount of ethanol to granulate, dry, granulate, add magnesium stearate, and then pack into gelatin capsules to obtain the capsule for treating senile dementia. The capsules are taken orally, 3 times a day, 1-3 capsules each time.

Embodiment 5 of the present invention: the application of total alkaloids of Dendrobium nobile:

Take 100-5000 mg of Dendrobium nobile total alkaloid extract, 1-10 grams of dextrin or sucrose, appropriate amount of flavoring agent and sweetener, and mix the extract evenly with sucrose/dextrin, flavoring agent and sweetener , add appropriate amount of water or ethanol to make soft material, sieve and granulate, dry, granulate, sub-package to get the granule for treating senile dementia. The granules are taken orally, 3 times a day, 5-10 grams each time.

Embodiment 6 of the present invention: the application of total alkaloids of Dendrobium nobile:

Take 100-2000 mg of total alkaloid extract of Dendrobium nobile, 100-2000 mg of polyethanol or soybean oil, 3-10 mg of suspending agent, 3-10 mg of emulsifier, 50-100 mg of gelatin, 10-30 mg of glycerin, 50-100 mg of purified water and an appropriate amount of preservatives, put gelatin in a sol tank, add purified water, heat to dissolve at 70°C, add glycerin and preservatives, stir evenly, remove air bubbles in a vacuum, keep it warm, and mix The extract is uniformly mixed with polyethanol/soybean oil, emulsifier, and suspending agent, and then placed in a rotary capsule press with the prepared gelatin, compressed into a soft capsule, shaped, and dried to obtain a soft capsule for treating senile dementia. The soft capsule is taken orally, 3 times a day, 1-3 capsules each time.

Embodiment 7 of the present invention: the application of total alkaloids of Dendrobium nobile:

Take 10-200 mg of Dendrobium nobile total alkaloid extract, 20-40 mg of polyethylene glycol 4000, and an appropriate amount of methyl silicone oil, add water to the extract to make a uniform paste, and then add the molten base (polyethylene glycol 4000) Liquid, heated and melted into a clear liquid, poured into the preheated pill dropper, controlled the dripping temperature and speed, dripped into the methyl silicone oil condensate, sucked up the condensate after forming pills, collected the dripping pills, and put them in a dryer In the medicine, the drop pill for treating senile dementia can be obtained. The dropping pill is taken orally, 3 times a day, 10-50 mg each time.

Embodiment 8 of the present invention: application of total alkaloids of Dendrobium nobile: take 200-5000 mg of total alkaloids extract of Dendrobium nobile, 1.5 grams of carboxymethyl cellulose sodium, 0.1 gram of sodium saccharin, appropriate amount of correctives, antiseptic Appropriate amount of agent and 100 ml of purified water, disperse sodium carboxymethyl cellulose in hot water, cool, then mix with aqueous suspension containing extract, sodium saccharin, flavoring and preservatives, and prepare the solution into the It needs volume and is mixed evenly, and then it is subpackaged after sterilization to obtain the syrup for treating senile dementia. The syrup is taken orally, 3 times a day, 5-10 ml each time.

Embodiment 9 of the present invention: application of total alkaloids of Dendrobium nobile: get 50-2000 mg of total alkaloids extract of Dendrobium nobile, 40-80 mg of compressible sucrose, 0.1-0.3 mg of magnesium stearate, extract After being sieved, it is evenly mixed with compressible sucrose and magnesium stearate, and pressed into tablets with a punching device to obtain a sublingual lozenge for treating senile dementia. The preparation is taken orally, 3 times a day, 1 tablet each time.

Embodiment 10 of the present invention: application of total alkaloids of Dendrobium nobile: get 10-2000 mg of total alkaloids extract of Dendrobium nobile, add water to 1000ml, add 0.5% gac, keep pH value 7.0, heat and boil slightly for 15 minutes, Cool, filter, add water for injection to the full amount, pack in cans, sterilize at 115°C for 30 minutes, refrigerate for 48 hours, filter, concentrate the filtrate, spray dry, and subpackage to obtain the injection for treating senile dementia. The preparation is for injection, 3 times a day, 5-10 ml each time.

Claims (5)

1. the application of Herba Dendrobii total alkaloid in the medicine of preparation treatment senile dementia is characterized in that: be crude drug with the Herba Dendrobii total alkaloid, process soft capsule as follows: get 100～2000 milligrams of Herba Dendrobii total alkaloid extracts, gather ethanol or 100～2000 milligrams in Oleum Glycines, 3～10 milligrams of suspending agents, 3～10 milligrams of emulsifying agents and 50～100 milligrams in gelatin, 10～30 milligrams of glycerol, 50～100 milligrams of purified water and antiseptic are an amount of; Gelatin is placed glue pot, add purified water, 70 ℃ of following heating make dissolving; Add glycerol and antiseptic, stir, insulation is left standstill after the vacuum removal of air bubbles; In proportion with extract with gather ethanol/Oleum Glycines, emulsifying agent, suspending agent mix homogeneously; Place rotation to press the capsule machine with the gelatin for preparing again, be pressed into soft capsule, typing; Drying promptly gets.

2. according to the application of the said Herba Dendrobii total alkaloid of claim 1, it is characterized in that: described Herba Dendrobii total alkaloid is the lipotropy total alkaloids constituents that from Dendrobium nobile, extracts.

3. according to the application of claim 1 or 2 said Herba Dendrobii total alkaloid, it is characterized in that: the method for preparing of said Herba Dendrobii total alkaloid is: depletion hairpin Herba Dendrobii medical material, be ground into coarse powder, and add ethanol extraction 3 times; Reclaim ethanol to there not being the alcohol flavor, transfer PH to 3～4, filter with acid solution; Filtrating is transferred PH to 10 with aqueous slkali, and an amount of chloroform extraction of reuse 5～8 times is collected chloroform layer; Reclaim chloroform, drying promptly gets.

4. according to the application of the said Herba Dendrobii total alkaloid of claim 3, it is characterized in that: the method for preparing of said Herba Dendrobii total alkaloid is: depletion hairpin Herba Dendrobii medical material is ground into coarse powder, the alcohol reflux of adding 95% 3 times; Reclaim ethanol to there not being the alcohol flavor, transfer PH to 3～4, filter with hydrochloric acid; Filtrating is transferred PH to 10 with 10% ammonia, and an amount of chloroform extraction of reuse 5～8 times is collected chloroform layer; Reclaim chloroform, drying promptly gets.

5. according to the application of the said Herba Dendrobii total alkaloid of claim 4, it is characterized in that: lipotropy total alkaloid contents >=60% in the said Herba Dendrobii total alkaloid.