CN113577240B

CN113577240B - Pharmaceutical composition, preparation method and application thereof

Info

Publication number: CN113577240B
Application number: CN202111025743.0A
Authority: CN
Inventors: 郭增光; 王言玉; 田宝祥; 李秀民
Original assignee: Xiangyu Pharmaceutical Co ltd
Current assignee: Xiangyu Pharmaceutical Co ltd
Priority date: 2021-09-02
Filing date: 2021-09-02
Publication date: 2024-01-30
Anticipated expiration: 2041-09-02
Also published as: CN113577240A

Abstract

The invention provides a pharmaceutical composition, a preparation method and application thereof, and relates to the technical field of traditional Chinese medicine preparations. The glutathione in the pharmaceutical composition has the effects of detoxification, oxidation resistance and growth promotion, the peptidoglycan has the effect of enhancing immunity, the vitamin E also has the effect of oxidation resistance, and the pharmaceutical composition prepared by the three components combined with fermentation of Cordyceps powder, ganoderma lucidum and medlar can effectively improve the effects of tonifying liver and kidney, replenishing qi to invigorate spleen, tonifying kidney to strengthen yang, soothing nerves and enriching blood through the synergistic combination of the raw materials, has remarkable effects in improving immunity, strengthening yang, improving memory, enhancing endurance, improving sleep, reducing blood fat, resisting arrhythmia and the like, and can effectively relieve side effects brought to patients in the radiotherapy and chemotherapy treatment process.

Description

Pharmaceutical composition, preparation method and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a pharmaceutical composition, a preparation method and application thereof.

Background

Cancer is a major class of malignant diseases that currently pose serious threat to human health and lack effective treatments. The disease and cardiovascular diseases become two main fatal diseases of human beings at present, and are the main death causes in most parts of the world. Currently the main treatments for cancer include radiation therapy (commonly known as radiotherapy), chemotherapy (commonly known as chemotherapy), and surgical therapies. The operation therapy is suitable for non-metastatic solid tumors, and the middle and late stage tumors need to be treated by combining radiotherapy and chemotherapy.

However, in the radiotherapy and chemotherapy treatment process, serious side effects are often brought to patients, and traditional Chinese medicine has shown a certain clinical curative effect in the aspect of preventing and treating toxic and side effects caused by radiotherapy and chemotherapy. For insomnia, listlessness, dizziness, palpitation, insomnia and the like caused by radiotherapy and chemotherapy, the Chinese medicine considers that the Chinese medicine belongs to deficiency of liver and kidney and weakness of spleen and qi, and thus, the Chinese medicine is used for treating tonifying traditional Chinese medicines such as tonifying kidney, tonifying qi, nourishing blood and the like. In the aspect of single traditional Chinese medicine, ginseng, angelica, astragalus, donkey-hide gelatin and medlar are commonly used. In compound research, it is often used in the form of a Yougui pill. However, the above disclosed traditional Chinese medicine has larger prescription and more medicine. The traditional Chinese medicine compound with smaller prescription and definite curative effect is lacking.

Therefore, research and development of a pharmaceutical composition with the effects of tonifying liver and kidney, replenishing qi to invigorate spleen, tonifying kidney to strengthen yang, soothing nerves and replenishing blood, capable of effectively relieving side effects in the radiotherapy and chemotherapy treatment process of cancer, and having remarkable lifting effects on improving immunity, strengthening yang, improving memory, enhancing endurance, improving sleep, reducing blood fat, resisting arrhythmia and the like becomes necessary and urgent.

In view of this, the present invention has been made.

Disclosure of Invention

The first aim of the invention is to provide a pharmaceutical composition, which can effectively improve the effects of tonifying liver and kidney, replenishing qi and strengthening spleen, tonifying kidney and strengthening yang, soothing nerves and replenishing blood of a product through the synergistic combination of the raw materials, has obvious effects in improving immunity, strengthening yang, improving memory, enhancing endurance, improving sleep, reducing blood fat, resisting arrhythmia and the like, and can effectively relieve side effects brought to patients in the radiotherapy and chemotherapy treatment process.

A second object of the present invention is to provide a method for preparing a pharmaceutical composition.

The third object of the invention is to provide an application of a pharmaceutical composition, which can be widely applied to the preparation process of auxiliary therapeutic drugs for radiotherapy and chemotherapy of cancers.

In order to achieve the above object of the present invention, the following technical solutions are specifically adopted:

the invention provides a pharmaceutical composition which is mainly prepared from fermented cordyceps fungus powder, ganoderma lucidum, medlar, glutathione, vitamin E and peptidoglycan.

Further, the pharmaceutical composition comprises, in parts by mass:

200-300 parts of fermented cordyceps fungus powder, 200-300 parts of lucid ganoderma, 30-80 parts of medlar, 10-30 parts of glutathione, 3-8 parts of vitamin E and 8-12 parts of peptidoglycan.

Further, the pharmaceutical composition comprises, in parts by mass:

250 parts of fermented cordyceps fungus powder, 250 parts of lucid ganoderma, 50 parts of medlar, 20 parts of glutathione, 5 parts of vitamin E and 10 parts of peptidoglycan.

The preparation method of the pharmaceutical composition provided by the invention comprises the following steps:

respectively extracting fermented Cordyceps powder, ganoderma and fructus Lycii, and preparing into paste fermented Cordyceps powder extract, paste Ganoderma extract and paste fructus Lycii extract; and then mixing the extracts to obtain a material A, uniformly mixing the material A with glutathione, vitamin E and peptidoglycan, and then drying and granulating to obtain the pharmaceutical composition.

Further, the extraction method of the fermented cordyceps sinensis bacterial powder comprises the following steps: adding the fermented cordyceps sinensis powder into ethanol for leaching, and then concentrating under reduced pressure to obtain a pasty fermented cordyceps sinensis powder extract;

preferably, the extraction method of the fermented cordyceps sinensis bacterial powder comprises the following steps: extracting the fermented cordyceps sinensis bacterial powder twice by using ethanol with the concentration of 60%, wherein the feed liquid ratio of each extraction is 1: 7-8, wherein the time for each extraction is 10-12 h; then merging the leaching solutions, and concentrating the merged leaching solution under reduced pressure to obtain fluid extract with the relative density of 1.20-1.25, thus obtaining pasty fermented cordyceps sinensis bacterial powder extract;

preferably, the combined extract is concentrated under reduced pressure at a temperature of 70-75 ℃.

Further, the fermented Cordyceps powder is one of fermented Cordyceps powder Cs-4, fermented Cordyceps powder Cs-C-Q80, cordyceps, sub-fragrant Cordyceps, cordyceps sinensis, cordyceps militaris, cordyceps sinensis, paederia, cordyceps sinensis, and preferably fermented Cordyceps powder CS-4.

Further, the extraction method of the ganoderma lucidum comprises the following steps: decocting Ganoderma in water, and concentrating under reduced pressure to obtain pasty Ganoderma extract;

Preferably, the extraction method of the ganoderma lucidum comprises the following steps: the ganoderma lucidum is decocted twice by using water, and the feed liquid ratio of each decoction is 1: 7-8, wherein the time of each decoction is 1.5-2 h; then merging the decoctions, and concentrating the merged decoction under reduced pressure to obtain fluid extract with the relative density of 1.20-1.25, thus obtaining pasty ganoderma lucidum extract;

preferably, the temperature of the combined decoction is 70-75 ℃ after decompression concentration.

Further, the extraction method of the medlar comprises the following steps: extracting fructus Lycii with ethanol, and concentrating the extractive solution under reduced pressure to obtain pasty fructus Lycii extract;

preferably, the extraction method of the medlar comprises the following steps: extracting fructus Lycii twice with 70% ethanol, wherein the ratio of extractive solution is 1: 4-5, wherein the time for each extraction is 48-50 h; then combining the leaching solutions, and concentrating the combined leaching solutions under reduced pressure to obtain clear paste with the relative density of 1.20-1.25 to obtain pasty medlar extract;

Further, the temperature of the dry granulation is 60-80 ℃.

The invention provides an application of the pharmaceutical composition in preparing a cancer radiotherapy and chemotherapy auxiliary treatment drug.

Compared with the prior art, the invention has the beneficial effects that:

the invention provides a pharmaceutical composition which is mainly prepared from fermented cordyceps fungus powder, ganoderma lucidum, wolfberry fruit, glutathione, vitamin E and peptidoglycan. The glutathione in the pharmaceutical composition has the effects of detoxification, oxidation resistance and growth promotion, the peptidoglycan has the effect of enhancing immunity, the vitamin E also has the effect of oxidation resistance, and the pharmaceutical composition prepared by the three components combined with fermentation of Cordyceps powder, ganoderma lucidum and medlar can effectively improve the effects of tonifying liver and kidney, replenishing qi to invigorate spleen, tonifying kidney to strengthen yang, soothing nerves and enriching blood through the synergistic combination of the raw materials, has remarkable effects in improving immunity, strengthening yang, improving memory, enhancing endurance, improving sleep, reducing blood fat, resisting arrhythmia and the like, and can effectively relieve side effects brought to patients in the radiotherapy and chemotherapy treatment process.

The invention provides a preparation method of a pharmaceutical composition, which comprises the steps of firstly, respectively extracting fermented cordyceps fungus powder, lucid ganoderma and medlar to prepare pasty fermented cordyceps fungus powder extract, pasty lucid ganoderma extract and pasty medlar extract; and then mixing the extracts to obtain a material A, uniformly mixing the material A with glutathione, vitamin E and peptidoglycan, and then drying and granulating to obtain the pharmaceutical composition. The preparation method has the advantages of simple preparation process, easy operation and suitability for industrial mass production.

The pharmaceutical composition provided by the invention can be widely applied to the preparation process of auxiliary therapeutic drugs for radiotherapy and chemotherapy of cancers.

Detailed Description

The technical solutions of the present invention will be clearly and completely described in connection with the embodiments, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

According to one aspect of the invention, a pharmaceutical composition is prepared mainly from fermented Cordyceps powder, ganoderma lucidum, fructus Lycii, glutathione, vitamin E and peptidoglycan.

In a preferred embodiment of the present invention, the pharmaceutical composition comprises, in parts by mass:

In the invention, the drug effect of the pharmaceutical composition is further optimized by further adjusting and optimizing the dosage proportion of the raw materials of each component.

According to one aspect of the present invention, a method for preparing the above pharmaceutical composition comprises the steps of:

In a preferred embodiment of the invention, the extraction method of the fermented cordyceps sinensis bacterial powder comprises the following steps: adding the fermented cordyceps sinensis powder into ethanol for leaching, and then concentrating under reduced pressure to obtain a pasty fermented cordyceps sinensis powder extract;

As a preferred embodiment, the fermented cordyceps sinensis powder is extracted by ethanol, so that the extraction yield of the effective components such as adenosine, cordycepic acid and the like in the fermented cordyceps sinensis powder is higher compared with the extraction mode of adopting water as an extraction medium in the prior art, and meanwhile, the reduced pressure concentration temperature is only 70-75 ℃, and the economic efficiency is better compared with the extraction temperature of 95-100 ℃ of the existing fermented cordyceps sinensis powder.

In a preferred embodiment of the present invention, the fermented Cordyceps powder is one of fermented Cordyceps powder Cs-4, fermented Cordyceps powder Cs-C-Q80, cordyceps, sub-fragrant Cordyceps, cordyceps sinensis, cordyceps militaris, cordyceps sinensis, paederia, guizhou Cordyceps, cordyceps sinensis, and Cordyceps sinensis, preferably fermented Cordyceps powder CS-4.

In a preferred embodiment of the present invention, the method for extracting ganoderma lucidum comprises: decocting Ganoderma in water, and concentrating under reduced pressure to obtain pasty Ganoderma extract;

In a preferred embodiment of the present invention, the method for extracting wolfberry fruit comprises: extracting fructus Lycii with ethanol, and concentrating the extractive solution under reduced pressure to obtain pasty fructus Lycii extract;

In a preferred embodiment of the present invention, the temperature of the dry granulation is 60 to 80 ℃.

According to one aspect of the invention, the application of the pharmaceutical composition in preparing a cancer radiotherapy and chemotherapy auxiliary treatment drug is provided.

The technical scheme of the invention will be further described with reference to examples.

Examples 1 to 5

A pharmaceutical composition comprising, in parts by mass:

the preparation method of the pharmaceutical composition comprises the following steps:

(1) Extracting fermented Cordyceps powder with 60% ethanol for 2 times, adding 8 times of 60% ethanol each time for 12 hr, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, and concentrating under reduced pressure to obtain fluid extract with relative density of 1.20-1.25;

(2) Decocting Ganoderma in water for 2 times and 8 times of water each time, decocting for 2 hours for the first time and 1.5 hours for the second time, mixing decoctions, filtering, and concentrating the filtrate under reduced pressure to obtain fluid extract with relative density of 1.05-1.10;

(3) Extracting fructus Lycii with 70% ethanol for 2 times, adding 5 times of 70% ethanol each time for 48 hr, mixing extractive solutions, filtering, recovering ethanol from filtrate under reduced pressure, and concentrating under reduced pressure to obtain fluid extract with relative density of 1.20-1.25;

(4) Mixing the above fluid extracts, mixing, adding steviosin, and mixing;

(5) Taking the fluid extract, glutathione, peptidoglycan, vitamin and lactose, granulating, drying, and preparing 1000g of drug particles, wherein the requirement of the drug particles is that the sum of particles passing through a No. 2 sieve and a No. 5 sieve is not more than 15 percent.

Comparative example 1

A pharmaceutical composition comprising, in parts by mass:

250 parts of fermented cordyceps sinensis powder, 250 parts of ganoderma lucidum and 50 parts of medlar;

(4) Mixing the above extracts, and drying to obtain 1000g of medicinal composition.

Experimental example 1 pharmacodynamic test of the reinforcing immunity of Yishen particles composed of different prescriptions

1. Test materials:

1.1 animals: pure HIN healthy mice are purchased from medical laboratory animal fields in Guangdong province, the mice are 5-6 weeks old, the weight is 20+/-2 g, and the male and female animals are about half, and are randomly grouped.

1.2 drugs:

the pharmaceutical composition (formula 1) prepared in comparative example 1 was diluted with distilled water to 43.2mg/ml before the test.

The pharmaceutical composition prepared in example 5 (formulation 2) was diluted with distilled water to 43.2mg/ml before testing.

Cyclophosphamide is formulated in distilled water to 1mg/ml before use.

Before the levamisole salt is used, distilled water is used for preparing 1mg/ml.

2. The test method comprises the following steps:

2.1 grouping of animals and dosing:

normal control group: gastric feeding distilled water 0.5 ml/field for 10 consecutive days;

(prescription 1) group: 1.08 g/kg/day, with the doses all contained in 0.5ml for 10 consecutive days;

(prescription 2) group: 1.08 g/kg/day, with the doses all contained in 0.5ml for 10 consecutive days;

levamisole group: gastric feeding mice on 5 days 1, 2, 3, 8, 9 were given distilled water at 25 mg/kg/day levamisole for the remaining 5 days;

cyclophosphamide group: the method for feeding distilled water is the same as that of a normal control group, and 10mg/kg of cyclophosphamide is injected into the abdominal cavity of the mice on the 8 th day according to the weight of the day;

ring + prescription 2 group: the administration method is the same as that of the prescription 2 group, and the administration method of cyclophosphamide is the same as that of the cyclophosphamide group;

ring + levamisole group: the administration method is the same as that of levamisole, and the administration method of cyclophosphamide is the same as that of cyclophosphamide group.

2.2 measurement method:

(1) The spleen cell quantitative hemolysis spectrophotometry is slightly improved:

the method is used for measuring hemoglobin (expressed by OD value) released by the lysis of erythrocytes by antibodies produced by a certain number of spleen B lymphocytes (referred to as spleen cells for short) to reflect the humoral immunity function of the body.

On day six, mice in each group were intraperitoneally injected with 0.4 ml/dose of 5% Sheep Red Blood Cells (SRBC) (equivalent to 4 x 108 SRBC/dose), the eyeballs were collected for exsanguination on day 11, the mice were sacrificed by neck pulling, spleens were taken to prepare spleen cell suspensions, the spleen cells were washed 2 times with cold Gey's solution, and the cell numbers were adjusted to 5 x 106/ml. 1ml of spleen cell suspension, 1% SRBC1ml and 1:20 fresh guinea pig complement (absorbed by SRBC) were added to each tube, thoroughly mixed, placed in a 37℃water bath for 1 hour, and after removal, cold Gey's solution was added to each tube to terminate the reaction. The absorbance supernatant after centrifugation was read for optical density (OD value) at 413nm with a 721-type spectrophotometer.

(2) T lymphocyte nonspecific lipase (ANAE) activity assay (ginger modification method):

the method is used for detecting the T lymphocytes in the mature period of peripheral blood so as to reflect the cellular immune function of the organism.

On test day 11, a tail blood smear of the mice is taken, quickly dried and immersed in an incubation liquid (a hexaazo fuchsin solution and a 2% alpha-naphthylacetate solution are added in proportion by using an M/15PH7.6 phosphate buffer solution, the final PH is 6.4) for 3 hours, the glass slide is taken out, sediment is washed by water, filter paper is sucked dry, and then 1% malachite green is counterstained for 5-10 seconds, washed with water, dried and inspected by an oil mirror. Each mouse was examined for 1 blood sheet, 100 lymphocytes were counted, and the percent of ANAE (+) T lymphocytes was determined.

3. Experimental results:

3.1 Effect of pharmaceutical composition on production of normal mouse spleen cell antibody:

the results are shown in Table 1, and the hemolysis OD values of the prescription 1 group and the prescription 2 group are respectively compared with that of the control group, and are both P & gt 0.05, which indicates that the three doses of the pharmaceutical composition have no obvious effect on the generation of normal mouse spleen cell antibody, and levamisole serving as a positive control medicine has no obvious effect.

Table 1 effect on normal mouse spleen cell hemolysis OD values:

note that: the methods of administration of this table, as well as of the other groups in each table below, are detailed in the animal groups and administration.

3.2 effects of pharmaceutical compositions on the production of antibodies by splenocytes of immunocompromised mice:

the results are shown in Table 2, the hemolysis OD values of the group 2 of the ring + pharmaceutical composition are respectively P < 0.001 compared with those of the cyclophosphamide group, and P is more than 0.05 compared with the normal group, which suggests that the pharmaceutical composition of the group 2 of the pharmaceutical composition has the effect of obviously enhancing the antibody production of the spleen cells of the immune-restricted mice and can enable the spleen cells to approach the normal level, and the pharmaceutical composition of the formula 1 has no obvious influence on the index.

TABLE 2 Effect on hemolysis OD values of splenocytes of immunocompromised mice

Note that: comparison of cyclophosphamide with normal group P < 0.001 indicates that immune suppression model was established.

3.3 effects of pharmaceutical composition on% of normal mouse peripheral blood ANAE (+) lymphocytes:

the results are shown in Table 3, and the results show that the% of ANAE (+) lymphocytes in the prescription 1 group and the prescription 2 group of the pharmaceutical composition are respectively compared with the normal group, and are both P & gt 0.05, which suggests that the pharmaceutical compositions of the two prescriptions have no obvious effect on the% of ANAE (+) lymphocytes in peripheral blood of normal mice, and levamisole serving as a positive control drug has no obvious effect.

Table 3 effect on% of normal mouse peripheral blood ANAE (+) lymphocytes:

3.4 effects of pharmaceutical composition on% of the immune-suppressed mice peripheral blood ANAE (+) lymphocytes:

the results are shown in Table 4, where the% ANAE (+) lymphocytes from the group 2 prescribed for the ring + pharmaceutical composition are compared to the cyclophosphamide group and P < 0.001, and then compared to the normal group, P > 0.05, suggesting that the pharmaceutical composition from the group 2 prescribed has an effect of significantly enhancing the% of ANAE (+) lymphocytes in the peripheral blood of the immunosuppressed mice and can be brought to near normal levels, whereas the pharmaceutical composition from the group 1 prescribed has no effect on this.

Table 4 effect on% of immune-suppressed mouse ANAE (+) lymphocytes:

note that: comparison of cyclophosphamide group with normal group P < 0.001 indicates that immune suppression model is established.

4. Discussion of results:

the prescription 2 of the pharmaceutical composition has the effects of obviously enhancing the generation of antibodies by spleen cells of immunocompromised mice (cyclophosphamide model) and improving the percentage of peripheral blood T lymphocytes, can prevent and treat the inhibition of cyclophosphamide on immune cells so as to improve humoral immunity and cellular immunity functions in a low state, and can enable the cells to approach to normal level to express the immunopotentiation and immunoregulation effects. The results suggest that the immunopotentiation and immunoregulation effects of the pharmaceutical composition are closely related to the immune function state of the organism, and are related to the theory of traditional Chinese medicine: the concept of "deficiency tonifying and impairment benefiting" is consistent. The pharmaceutical composition of prescription group 1 had no significant effect on both normal and immunocompromised mice.

Levamisole is a positive immunopotentiator, and is used as a positive control drug in the test. Experimental results show that the pharmaceutical composition has the effects of obviously enhancing humoral immunity and cellular immunity of the mice with the immunity being inhibited, and can enable the mice to approach to the normal level, but has no obvious influence on the normal mice. The test result is consistent with the characteristic of the immunopharmacological action of levamisole.

Experimental example 2 pharmacodynamic test of Yishen granule with kidney-tonifying and yang-strengthening effects composed of different prescriptions

1. Experimental materials:

1.1 test drug:

(1) Yishen granule: comparative example 1 (formulation 1) and example 5 (formulation 2).

(2) Man Bao capsule: tianjin is a Lisheng pharmaceutical factory product.

(3) Jinkui kidney qi pill (honeyed pill): the product of the pharmaceutical factory in the city of Buddha.

1.2 main reagents:

(1) Serum testosterone radioimmunoassay kit: tianjin Depu (DPC) Biotechnology and pharmaceutical products Co., ltd.

(2) Hydrocortisone injection: hubei pharmaceutical factory product.

1.3 experimental animals: SD rats, NIH rats were purchased from medical laboratory animal sites in Guangdong province.

1.4 main instruments: FT-613 automatic counting 125I meter.

2. Test methods and results:

2.1 Effect on the accessory organs of immature Male mice

42 healthy male mice are selected, and the weight of the healthy male mice is 11-13g, and the healthy male mice are randomly divided into distilled water groups, positive medicine groups (Male treasure capsule groups), yishen granule prescription 1 groups and Yishen granule prescription 2 groups (which are 20 times of clinical dosages). Then, the medicines were administered by stomach irrigation (medicines were formulated at different concentrations, were administered at a weight of 0.2ml/10g, distilled water was administered in distilled water groups in equal amounts) 1 time a day for 7 consecutive days. Animals were sacrificed the next day after the last dose, prepuced glands, prostate and seminal vesicles were dissected, precisely weighed, organ indices calculated, and differences between groups were compared. The results are shown in Table 5.

Table 5 effect on the weight of accessory organs of immature male mice (x±sd):

note that: comparison with distilled water group: * P is less than 0.05; * P < 0.01 (the same applies below).

The results in Table 5 show that the group 2 of Yishen granule prescription has obvious weight increasing effect on the accessory organs of immature male mice, and the difference is very significant (P is less than 0.01) compared with the distilled water control group; however, the weight gain effect of the prescription 1 group is not obvious (P is more than 0.05). The male BAO capsule group mice only have the weight increasing effect on the prostate and seminal vesicles (P is less than 0.05), but the weight increasing effect on the prepuced glands is not obvious (P is more than 0.05).

2.2 effects on serum testosterone levels in immature male rats:

36 healthy male rats are selected, and are respectively and randomly grouped and administered by gastric lavage (the grouping and the administration method are the same, and are given according to the weight of 1ml/100 g), 1 time a day, and 12 continuous days. Fasted for 12 hours after the last administration, 0.6% sodium pentobarbital (7 ml/kg) was intraperitoneally anesthetized, the heart was bled to isolate serum, serum testosterone levels were measured by radioimmunoassay, and differences between the groups were compared. The results are shown in Table 6.

Table 6 effect on serum testosterone levels in immature male rats (x±sd):

the results in Table 6 show that both the Yishen granule prescription 1 and the prescription 2 can improve serum testosterone levels of immature male rats, and the differences are very remarkable (P < 0.01) compared with distilled water, and the effects are similar to those of Man Bao capsules (P > 0.05).

2.3 effects on castrated male rat accessory organs:

45 healthy male rats are selected, the weight of the healthy male rats is 200-220G, the pudendum skin is disinfected under shallow anesthesia by diethyl ether, the testes on both sides are removed, and 4 ten thousand U/patient is injected with penicillin G sodium by muscle after operation. Animals were randomly grouped on day 2 after castration surgery and given by gavage (normal control group was added as above for grouping and administration), 1 time daily for 10 consecutive days, weighing on day 2 after the drug, sacrificed, rapidly dissected seminal vesicles, prostate and prepuce glands, weighing, calculating organ indexes, and comparing the differences of the groups. The results are shown in Table 7.

Table 7 effect on castrated male rat accessory organs (x±sd):

as can be seen from the results of Table 7, the formulation 2 of Yishen granule has the effect of preventing prostate seminal vesicle atrophy of castrated male rats, the difference is significant (P < 0.05) compared with distilled water, the effect is similar to that of Menbao capsule (P > 0.05), the formulation 1 has the effect of insignificant (P > 0.05), both formulations of Yishen granule can prevent foreskin gland atrophy of castrated male rats, the difference is significant (P < 0.05) compared with control, and the effect is similar to that of Menbao capsule (P > 0.05).

2.4 effects on animals with kidney-yang deficiency:

52 healthy male mice were selected, weighing 20-22g, were administered by gastric lavage after random grouping (grouping and administration method were the same as in experiment 1), and simultaneously injected subcutaneously with hydrocortisone 25mg/kg (normal group administration of equivalent physiological saline) 1 time daily for 9 consecutive days. On day 10, mice were sacrificed after weighing, adrenal glands, thymus, spleen, prostate and seminal vesicles, prepuce glands and testes were dissected, weighed, organ indexes were calculated, and differences between the groups were compared. The results are shown in tables 8 and 9.

Table 8 effect on body weight, adrenal glands, thymus and spleen of kidney-yang deficient mice (X soil SD):

the results in Table 8 show that the weight of normal mice increases by 6.21g on average after 9 days, the weight of mice in yang deficiency model caused by hydrocortisone is obviously reduced (P < O.01), and the Yishen granule prescription 1 and prescription 2 can both prevent weight reduction, and compared with distilled water, the difference is very obvious (P < 0.01), and the effect is similar to that of kidney qi pill. Table 8 shows that group 1 of Yishen granule can prevent mice from atrophy of adrenal gland caused by hydrocortisone, and compared with distilled water group, the difference is significant (P < O.05-0.01), but the effect of preventing thymus and spleen atrophy is not obvious (P > 0.05); the effect of prescription 2 on preventing adrenal atrophy is not obvious (P > 0.05), but on preventing the rhombic shrinkage of thymus and spleen is obvious (P < 0.05-0.01).

Table 9 effect on kidney-yang deficiency mice testes, prostate and seminal vesicle glands, prepuce glands (X soil SD):

the results in Table 9 show that hydrocortisone can cause atrophy of the male mice' appendages (P < 0.01), while both prescription groups of the particles can prevent atrophy of the prostate and seminal vesicles, compared with distilled water groups, the difference is very remarkable (P < 0.01), and the prescription 2 group has obvious trend better than the prescription 1 group, can prevent atrophy of the prepuce glands (P < 0.01), and has similar effect as kidney qi pills; the group 1 of prescriptions had no obvious effect on preventing atrophy of the prepuce glands (P > 0.05).

3. The small knot: the results of this study showed that:

the Yishen granule prescription 2 has obvious strengthening effect on male animal reproductive system compared with the prescription 1. The Yishen granule prescription 2 has weight increasing effect on the accessory organs of immature male mice, has atrophy preventing effect on the accessory organs of yang deficiency model mice and castrated male rats, and can improve the serum testosterone level of immature male mice, so that the Yishen granule can promote the development of the accessory organs of male animals and improve the androgen level in the bodies of male animals, and has androgen-like effect. The prescription 2 of the Yishen granule has strengthening effect on male reproductive system, and is consistent with clinical kidney-tonifying and yang-strengthening effects.

The Yishen granule prescription 2 has the prevention and treatment effect on male mice yang deficiency caused by hydrocortisone: kidney yang deficiency is associated with hypofunction of the adrenal cortex system, reproductive system and immune system, whereas the Yishen granule of prescription 2 has significant advantages over the Yishen granule of prescription 1 in preventing atrophy of adrenal gland, accessory organ, thymus and spleen of mice model of kidney yang deficiency.

Experimental example 3 effects of different prescription compositions of the particles on mice Hb and RBC with acute blood loss anemia:

1. medicament:

1.1 Yishen granule (prescription 1): the pharmaceutical composition prepared in comparative example 1 (formulation 1) was diluted with distilled water to 16mg/0.3ml before the experiment.

1.2 Yishen granule (prescription 2): the pharmaceutical composition prepared in example 5 (formulation 2) was diluted with distilled water to 16mg/0.3ml prior to the experiment.

1.3 Xuebao capsules are produced by Guangzhou Chen Liji pharmaceutical factories and are formulated into suspensions with a concentration of 1.5mg/0.3ml with distilled water.

2. Animals: pure NIH healthy mice and feeds thereof are purchased from animal houses of Guangzhou Chinese medical college, the ages of the mice are 5-6 weeks, the weights of the mice are 18-20g, the male and female animals are about half, and the mice are randomly divided into an experimental prescription 1 group, a prescription 2 group, a Xuebao control group and a blank control group, and the total number of the mice is 4.

3. Acute hemorrhagic anemia model: the procedure described in the literature was followed by a glass capillary bleed of 0.5ml through the orbital vein of the mouse to create anemia. And taking a small amount of blood 1 day (via the tail tip) before and after blood loss to measure hemoglobin (Hb) and Red Blood Cells (RBC), and observing anemia after blood loss as objective indexes for evaluating modeling effect and drug effect.

4. Anaemia mice Hb and RBC recovery assay: the animals of the 4 groups are respectively irrigated with a prescription 1, a prescription 2, a Xuebao suspension and distilled water (0.3 ml/10 g) once daily, wherein the dosage of the prescription 1 and the 2 groups of Yishen granules is 20 times that of an adult. Animals were fed and drinking water freely during the test period, drenched for 7 days, and then small amounts of blood were collected from the mouse tail tips to measure Hb and RBC, and their changes were observed.

HB and RBC assay Hb assay was performed by Sha Lishi method and RBC assay was performed by photoelectric turbidimetry.

6. Statistical analysis: the Hb and RBC self comparisons before and after blood loss and before and after dosing of each group were performed by paired t-test, while the Hb and RBC post blood loss decrease values and post dosing increase values between each group were compared by anova and by pairwise comparison.

7. Results:

7.1 changes in Hb and RBC in mice before and after acute blood loss are shown in table 10.

Table 10 comparison of mouse Hb (g/L) and RBC (X1012/L) (X.+ -. SD) before and after blood loss:

* : p < 0.05 compared with that before blood loss. * *: p < 0.001 compared to before blood loss.

Delta: the difference between Hb and RBC reduction values for each group was not significant (P > 0.05). The results in table 10 show that Hb and RBC decrease very significantly in each group after blood loss, while the difference between the blood loss levels was not significant.

7.2 effects of Yishen particles on mice with anemia Hb (g/L) and RBC, the results are shown in Table 11 and Table 12.

Table 11 comparison (x±sd) of Hb (g/L) in mice with anemia before and after dosing:

* : p <0.05 compared to pre-dose. * *: p <0.001 compared to pre-dose. Delta: p <0.05 compared with the blank group. Delta delta: p <0.001 compared to the placebo group. And ∈: p <0.05 compared to the blood control group.

The statistical comparison of table 11 shows a significant rise in Hb (P < 0.001-0.05) after dosing of both prescription 1, 2 and control groups, with prescription 1, 2 being more pronounced. The difference between the prescription 1, the prescription 2 and the blank group is obvious (P < 0.01-0.05), the prescription 1 and the blood control group are also obviously sensitive, and the difference between the different prescription groups and the blood control group and the blank control group can not be displayed.

Table 12 comparison (x±sd) of RBC (X1012/L) in anaemic mice before and after dosing:

* *: p <0.001 compared to pre-dose. Delta: the difference between the increase values of each group was not significant (P > 0.05).

8. Discussion:

8.1 modeling of acute hemorrhagic anemia: from the change conditions of Hb and RBC before and after blood loss of the experimental animals, the model of the blood loss anemia achieves the expected purpose, and the comparison result (P > 0.05) of the blood loss degree between groups indicates that the blood loss degree of the groups is basically consistent, which provides a comparable basis for further observing the blood replenishing effect of the medicine.

8.2 recovery effect of Yishen granule (prescription 2) on blood loss anemia mouse Hb: experimental results show that Hb of each group of mice is obviously increased (P < 0.001-0.05) after 7 days of administration, wherein the prescription 1 and 2 groups of Yishen particles are particularly obvious, and comparison of the increase values of each group shows that the difference between the prescription 1 and 2 groups of Yishen particles and a distilled water blank group is obvious (P < 0.001-0.05), which indicates that Yishen particles of the two prescriptions have the effect of improving Hb. But there was no significant difference between prescription 2 and prescription 1.

8.3 recovery of RBC from blood loss anemia mice with Yishen granule (prescription 2): the experimental results showed that RBCs in each group significantly increased (P < 0.001) 7 days after dosing, but there was no significant difference between the increase values in prescription 2 and prescription 1 groups, failing to show that Yishen particles (prescription 2) are better than prescription 1 groups.

8.4 nodules: in conclusion, the prescription 2 and the prescription 1 of the Yishen granule have certain blood replenishing effect, but have no obvious difference.

Experimental example 4 pharmacodynamic experiments of anti-stress and sedative effects of Yishen particles composed of different prescriptions

1. Materials and methods: NIH mice, 18-22g in weight, were supplied by the animal house of the Guangzhou Chinese medical college. Soda lime is produced by Guangdong Taishan chemical plant, lot number 900608. The pentobarbital sodium is imported and split-packed by chemical engineering laboratory in the city of bergamot, and has a batch number of 360901. The chlordiazepoxide tablet is produced by fourth pharmaceutical factory in Changzhou, jiangsu province, lot number 870728. The ginseng royal jelly oral liquid is the product of Beijing Tofeng pharmaceutical factory, batch number 930919.

Yishen granule is the same as before. The experiments were divided into groups 1 and 2, the pharmaceutical composition prepared in comparative example 1 (formula 1), the pharmaceutical composition prepared in example 5 (formula 2) and the clinical dosage of the pharmaceutical composition was 15 times that of an adult. A positive drug control group and a blank distilled water control group are additionally arranged.

Atmospheric hypoxia test: according to the improvement of the literature method, mice are randomly grouped and continuously filled with medicines for 11 days, the medicines are fasted for 1 time every day, water is not forbidden for 5 hours before the last administration, the mice are put into a 250m1 wide-mouth bottle after 1 hour of administration, the bottle is closed, 4g of soda lime is filled in the bottle, the edge of the bottle cap is coated with vaseline for sealing, and the time from closing the bottle to death due to hypoxia is recorded.

Swimming test: according to the improvement of the literature method, male mice are randomly grouped and continuously filled with medicines for 11 days, 1 time a day, and are fasted for 5 hours before the last administration, after the administration for 1 hour, the tail of the mice is loaded by a frog heart and a lead fuse wire, the weight of the mice is 10 percent of the weight of the mice, the mice are put into a bucket filled with water, the water depth is 20cm, the water temperature is 18 ℃, and the mice are recorded until the limbs of the mice cannot be scratched due to fatigue when the mice are put into water.

Pentobarbital sodium collaborative hypnotic test: according to the literature method, mice are combined with each other, randomly grouped, continuously filled for 9 days, 1 time a day, fasted before the last administration, and not forbidden for 5 hours, each mouse is intraperitoneally injected with sodium pentobarbital for 30mg/kg after 30 minutes of administration, and the number of mice with everlasting reflection disappeared within 15 minutes after injection is recorded.

2. Results and analysis:

2.1 Effect of Yishen particles on mice atmospheric hypoxia tolerance test:

table 13 effect of Yishen particles on mice normbaric hypoxia test:

comparison with control group: p <0.05,: p < 0.01 (the same applies below).

As can be seen from table 13, the formulation 1 and 2 of the Yishen granule can prolong the survival time of mice under normal pressure hypoxia, and the difference between the formulation 2 and the control group is significant. The prescription 2 of Yishen granule can improve the hypoxia tolerance of mice. The ginseng royal jelly also has the same effect.

2.2 Effect of Yishen particles on cold-resistant swimming test in mice:

table 14 effects of Yishen particles on the swimming test furrows of mice:

as can be seen from table 14, both prescription groups of the Yishen granule can prolong the swimming time of mice, and the effect of prescription 2 is better than that of prescription 1. Compared with the control group, the prescription 2 has obvious significance in difference, and the effect is similar to that of ginseng royal jelly.

2.3 Effect of Yishen particles on sodium pentobarbital collaborative hypnotic test in mice:

table 15 effect of Yishen granule on sodium pentobarbital co-hypnotic test in mice:

chi-square test, delta compared to control group: p <0.005, Δ: p <0.05.

As can be seen from table 15, both prescription groups of the Yishen granule increased the number of mice sleeping, and prescription 2 had better effect than prescription 1. The difference is statistically significant in group 2 compared to the control, indicating that group 2 of Yishen granule has synergistic sodium pentobarbital hypnotic effect.

3. The small knot: the results of the Yishen granule prescription 2 group in the tests of swimming, normal pressure anoxia and synergic hypnotic with pentobarbital sodium show that the medicine has the functions of resisting fatigue, resisting anoxia and hypnotizing, and provides a certain pharmacological basis for clinical indications of the medicine.

Experimental example 5 preliminary experimental study of the composition of different prescriptions according to the Yishen granule for enhancing the memory capacity of mice and resisting lipid peroxidation:

the rare Chinese medicines such as cordyceps sinensis, ganoderma lucidum and the like have the functions of enhancing the mechanical break function, regulating and controlling the immune response of organisms, resisting aging and the like, and are the magnifications in the Chinese medicine treasury. The experiment researches the influence of the Yishen granule with different prescription compositions on the memory and lipid metabolism of mice.

1. Materials and methods:

1.1 experimental animals: 85 Kunming mice were selected, each half of which had a body weight of 30-32g.

1.2 Yishen granule prescription 1 and prescription 2 are provided by Xiangyu pharmaceutical company.

1.3 animals grouping:

(1) Effects on memory function:

group A: animals 18, each mouse was perfused with physiological saline 0.2m1 daily for 8 days.

Group B: animals 16, each mouse was perfused with 0.2ml physiological saline daily for 8 days.

Group C: animals 16 animals were gavaged daily at 1.3 g/kg/day Yishen granule (prescription 1) for 8 days.

Group D: animals 19, each mouse was gavaged daily for 8 days at 1.3 g/kg/day Yishen granule (prescription 2).

Group E: animals 16 animals were perfused with 7.5 g/kg/day Naofukang per mouse daily for 8 days.

(2) Anti-lipid peroxidation study:

physiological saline group of 18 animals, and each mouse is infused with physiological saline 0.2m1/kg every day for 8 days.

Animals in prescription 1 group 16, each mouse was gavaged daily at 1.3 g/kg/day Yishen granule (prescription 1) for 8 days.

Animals in prescription 2 groups were 19, and each mouse was perfused with 1.3 g/kg/day Yishen granule (prescription 2) daily for 8 days.

The vitamin E group contains 16 animals, and each mouse is 2ml/0.2m1 vitamin E for gastric lavage for 8 days.

1.4 determination of memory function: the method is characterized in that a clear box device and a camera bellows device are used, a passive avoidance response of a mouse is used as an index to measure memory capacity, the camera bellows is illuminated by light, a copper grid is arranged at the bottom of the camera bellows to enable the animal to be shocked, a small hole is arranged between the two boxes for the mouse to pass through, the first day of test is to place the mouse in the camera bellows, the darkness of the mouse is utilized to record the residence time of the mouse before the four limbs of the mouse enter the camera bellows, namely, the entrance latency period is recorded, the animal is shocked (30V alternating current) immediately after the mouse enters the camera bellows, the mouse escapes to the camera bellows for 30 seconds, the mouse is taken out after the mouse escapes to the camera bellows, the same time is placed in the camera bellows on the next day, the re-entrance latency period is observed, the memory capacity is divided into three conditions according to the re-entrance latency period, the person who does not enter the camera bellows on the next day within 5 minutes after the shock is "memory slightly", the entrance latency period difference between the entrance latency period before the shock is less than 30 seconds is "memory slightly". Four groups of mice were tested 30 minutes prior to memory for B, C, D, E, and each mouse was perfused with 0.1ml/10g of 40% alcohol (100% assay alcohol diluted with saline) to observe and compare the adverse effects of the above-described traditional Chinese medicine and brain complex Kang Jie alcohol.

1.5 detection of scavenging superoxide radical:

the effects of the above Chinese medicines on biological oxidation and aging process of the organism are detected by taking the superoxide dismutase (SOD) activity in blood and the Lipid Peroxide (LPO) content of liver homogenate as indexes.

After the experiment is finished, four groups of mice are killed by head breaking, heparin anticoagulation whole blood is collected, livers are taken out, 1 gram of liver tissue is added with 2ml of physiological saline to prepare homogenate, the heart is separated for 10 minutes at 3000 revolutions per minute, the supernatant is taken for later use, the erythrocyte SOD activity is measured by adopting a pyrogallol autoxidation method, the LPO content in the liver homogenate is measured by using a TBA color development method and is converted into Malondialdehyde (MDA) content, wherein the hemoglobin content is measured by using a photoelectric colorimetry method, and the liver plasma protein content is measured by using a lowry's method.

2. Results:

2.1 effects on memory function:

(1) The normal saline control group (A group) was 8 "memory-equipped" mice, 44.4% and 1 "slightly memory-equipped" animals, 5.6% and 9 "memory-free" animals, 50.0% of the 18 animals tested.

(2) The physiological saline (group B) is added with alcohol, the number of the mice to be tested is 2, accounting for 12.5 percent,

the "slightly memorized" is 0, 14 cases with "no memory" account for 87.5%, the sum of the numbers of animals with "memory" and "slightly memorized" is taken as one group, the "no memory" is taken as the other group, and the difference between the two experimental groups of A, B is compared by using a card method statistics method, so that the two groups have significant difference (X2=5.6901, P < 0.02). The results suggest that oral administration of 40% alcohol 0.lml/10g body weight 30 minutes prior to memory testing can significantly impair the memory function of mice.

(3) Prescription 1 (group C) add alcohol: 19 mice were tested, 30 minutes after taking the same dose of alcohol, 9 "with memory" accounting for 56.2%, 3 "with slightly memory" accounting for 18.8%, and 12 of the two summed to 75.0%, this value being very significantly different from the statistical comparison of group B (x2=10.2857, p < 0.01). "No memory" 4 cases, accounting for 25.0%. The results indicate that the Yishen granule (prescription 1 group) has the function of improving the memory function of mice.

(4) Prescription 2 (group D) add alcohol: 19 mice were tested and were given the same dose of alcohol for 30 minutes, 16 cases were "with memory" and were 79.0%, "with memory" and were 2 cases were 10.5%, the sum of the two was 89.5%, this value was very significantly different (x2=17.7515, p < 0.01) compared to the group B, and the group "without memory" was 2 cases and was 10.5%. The results show that the Yishen granule (prescription 2 group) has the function of obviously improving and protecting the memory capacity of mice.

(5) Naofukang (group D) with alcohol, detecting 16 mice, 30 minutes after taking the same dose of alcohol, 15 cases with memory, accounting for 93.7%, this value being very significantly different from the statistical comparison of group B (x2=18.2593, p < 0.01). The group has 0 cases of slightly memorized and 1 case of no memory, and accounts for 6.3 percent, and the result shows that the positive control medicine can obviously improve the memory capacity of mice and improve the impaired memory function after being orally taken by cerebral rehabilitation.

Table 16 comparison of memory function test values for animals of each experimental group

2.2 effects on scavenging superoxide radical:

(1) Effect on superoxide dismutase (SOD) activity: biochemical measurements carried out after 8 days of gastric lavage of each drug respectively show that the SOD enzyme activity of the Yishen granule red blood cells of both prescriptions is obviously higher than that of a saline control group, compared with the saline control group, the SOD enzyme activity of the Yishen granule red blood cells of both prescriptions is increased by 25 percent (t=2.2583 and P is less than 0.05), the SOD enzyme activity of the Yishen granule red blood cells of both prescriptions is also increased by 19 percent, and the statistical difference exists between the vitamin E group and the saline group, and the results are shown in Table 17.

(2) As can be seen from liver homogenate LPO measurements for liver Lipid Peroxide (LPO), formulation 2 reduced the degree of liver lipid peroxidation, which was very significant compared to the saline control group (t=3.7338, p < 0.01), see table 17.

Table 17 comparison of erythrocyte SOD Activity and liver LPO content in animals of each experimental group

Comparison to saline control group: * P < 0.05, P < 0.01.

3. Discussion: amnesia, mental retardation, etc. are important symptoms of aging. Deficiency of kidney essence and insufficiency of essence and sea of essence are caused by deficiency of qi and blood and deficiency of heart and spleen. The experimental results show that both the Yishen granule prescription 1 and the prescription 2 can improve the memory function of experimental mice and improve the memory damage caused by alcohol, but the effect of the prescription 2 is obviously better than that of the prescription 1. Diseases and aging are caused by peroxidation of free radicals, and are currently recognized at home and abroad. The body gains oxygen by breathing to sustain life activities. Oxygen generates oxidation radicals, hydrogen peroxide radicals, and the like during the reduction process. These free radicals are harmful to the body, and excessive amounts can destroy cells and their important components, cause DNA mutations and denature proteins, enzymes, etc., thereby accelerating aging. Scavenging free radicals is therefore of general importance for delaying aging. And superoxide dismutase (SOD for short) can remove them. The experimental results show that the Yishen granule prescription 2 and the Yishen granule prescription 1 both have the functions of promoting SOD enzyme activity, improving the capability of an organism to remove oxygen free radicals generated in metabolism, reducing the peroxidation degree of liver lipid, and resisting aging, and have no obvious difference.

Experimental example 6 Effect of different prescriptions on rat blood lipid by Yishen granule

The experiment mainly observes the influence of Yishen particles composed of different prescriptions on normal rat blood fat, and provides an experimental basis for clinical application of the medicine.

1. Materials and methods:

1.1 laboratory animals 47 Wistar rats were male and had a weight of 160.+ -.10 g.

1.2 Chinese medicinal preparation, wherein the concentration of the extract of Yishen granule prepared according to prescription 1 and prescription 2 is 1.3g crude drug per 1 ml. Distilled water is used to prepare the required concentration before use.

1.3 animals grouping:

(1) Normal saline group, 10 rats, were infused with gastric normal saline lml/rat daily for 10 days.

(2) The prescription 1 group of rats was 13 rats, each filled with the Yishen granule (prescription 1) at 0.435 g/kg/day for 10 days.

(3) Prescription 2 group of rats 11 rats were perfused with the stomach Yishen granule (prescription 2) at a daily rate of 0.87 g/kg/day for a total of 10 days.

(4) Antominium group 13 rats were perfused with antominium at a daily rate of 0.2 g/kg/day for 10 days.

1.4 blood lipid measurement: before and on day 10 of administration, each group of rats was lightly anesthetized with diethyl ether, blood was collected from the tail, serum was isolated, and serum cholesterol concentration was measured by direct assay of trace serum total cholesterol. The two measurements were compared and the percent drop was determined according to the following formula:

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2. Experimental results

Experimental results show that the Yishen granule and antomine of both prescriptions have the effect of reducing the total cholesterol in the serum of rats. See table 18.

TABLE 18 Effect of Yishen particles on serum Total cholesterol in Normal rats

3. Conclusion of experiment: the test results indicate that the prescription 1 group and the prescription 2 group of the Yishen granule have certain blood lipid reducing effect and have no obvious difference.

Experimental example 7 preliminary experimental study of antiarrhythmic properties of Yishen granule with different prescription compositions:

the aim of the research is to observe the influence of Yishen particles composed of different prescriptions on the arrhythmia of mice induced by chloroform, and provide a certain experimental basis for clinical application of the medicine.

1. Materials and methods:

1.1 experimental animals 45 Kunming mice, male and female halves, body weight 30-35g.

1.2 Chinese medicinal preparation, YISHEN granule extract contains 1.3g crude drug per lml, and is diluted with distilled water to appropriate amount before use.

1.3 animals grouping:

(1) 10 mice were perfused with 0.2 ml/mouse physiological saline daily for 5 days.

(2) The prescription 1 group of mice were perfused with 0.43 g/kg/day Yishen granule (prescription 1) daily for 5 days.

(3) Prescription 2 groups 13 mice were gavaged daily at 0.86 g/kg/day Yishen granule (prescription 2) for 5 days.

(4) Propranolol group; 10 mice were perfused with the triamcinolone acetonide at 0.006 g/kg/day for 5 days.

1.4 chloroform induced arrhythmia in mice:

the mice of each group above were placed one by one in an inverted 500ml beaker containing 3-4ml chloroform cotton balls 2 hours after the 6 th day of the experiment, the split chest was immediately removed until respiratory arrest, and the incidence of ventricular fibrillation was visually examined.

2. Experimental results: experimental results show that the prescription 2 group has a certain effect of resisting the mice arrhythmia caused by chloroform, while the prescription 1 group has no obvious effect. See table 19.

Table 19 antagonism of particles of Yishen to ventricular fibrillation by inhalation of chloroform in mice:

3. conclusion of experiment:

the test results indicate that the prescription 2 group of Yishen granules has certain antiarrhythmic effect.

As can be seen from the data of the above experimental examples, the pharmaceutical composition prepared in example 5 of the present application has the following technical advantages compared with the pharmaceutical composition prepared in comparative example 1:

compared with the pharmaceutical composition prepared in the comparative example 1 (250 g of fermented cordyceps sinensis powder (Cs-4), 250g of ganoderma lucidum and 50g of medlar), the pharmaceutical composition prepared in the embodiment 5 of the application has obvious effects of improving immunity, strengthening yang, improving memory, enhancing endurance, improving sleep, reducing blood fat, resisting arrhythmia and the like.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.

Claims

1. The pharmaceutical composition is characterized by being prepared from fermented cordyceps fungus powder, ganoderma lucidum, wolfberry fruit, glutathione, vitamin E and peptidoglycan;

the medicine composition comprises the following raw materials in parts by mass: 200-300 parts of fermented cordyceps sinensis powder, 200-300 parts of lucid ganoderma, 30-80 parts of medlar, 10-30 parts of glutathione, 3-8 parts of vitamin E and 8-12 parts of peptidoglycan;

the fermented Cordyceps powder is fermented Cordyceps powder CS-4.

2. The pharmaceutical composition according to claim 1, characterized in that it consists of the following raw materials in parts by mass:

250 parts of fermented cordyceps sinensis powder, 250 parts of lucid ganoderma, 50 parts of medlar, 20 parts of glutathione, 5 parts of vitamin E and 10 parts of peptidoglycan;

the fermented Cordyceps powder is fermented Cordyceps powder CS-4.

3. A method of preparing a pharmaceutical composition according to claim 1 or 2, characterized in that the method of preparation comprises the steps of:

respectively extracting fermented Cordyceps powder, ganoderma and fructus Lycii, and preparing into paste fermented Cordyceps powder extract, paste Ganoderma extract and paste fructus Lycii extract; mixing the above extracts to obtain material A, mixing material A with glutathione, vitamin E and peptidoglycan, drying, and granulating to obtain pharmaceutical composition;

the fermented cordyceps sinensis bacterial powder is fermented cordyceps sinensis bacterial powder CS-4;

the extraction method of the fermented cordyceps sinensis bacterial powder comprises the following steps: extracting the fermented cordyceps sinensis bacterial powder twice by using ethanol with the concentration of 60%, wherein the feed liquid ratio of each extraction is 1: 7-8, wherein the time for each extraction is 10-12 h; then merging the leaching solutions, and concentrating the merged leaching solution at the temperature of 70-75 ℃ under reduced pressure to obtain fluid extract with the relative density of 1.20-1.25, so as to obtain paste-like fermented cordyceps sinensis bacterial powder extract;

the extraction method of the ganoderma lucidum comprises the following steps: the ganoderma lucidum is decocted twice by using water, and the feed liquid ratio of each decoction is 1: 7-8, wherein the time of each decoction is 1.5-2 hours; then merging the decoctions, and concentrating the merged decoction at a temperature of 70-75 ℃ under reduced pressure to obtain fluid extract with a relative density of 1.20-1.25, thus obtaining pasty ganoderma lucidum extract;

The extraction method of the medlar comprises the following steps: extracting fructus Lycii twice with 70% ethanol, wherein the ratio of extractive solution is 1: 4-5, wherein the time for each extraction is 48-50 h; then merging the leaching solutions, and concentrating the merged leaching solution at the temperature of 70-75 ℃ under reduced pressure to obtain fluid extract with the relative density of 1.20-1.25, thereby obtaining pasty medlar extract;

the temperature of the drying granulation is 60-80 ℃.

4. Use of the pharmaceutical composition according to claim 1 or 2 for the preparation of a medicament for adjuvant therapy of cancer chemoradiotherapy.