Use of a traditional Chinese medicine composition in the preparation of a medicine for treating organ fibrosis

Abstract

The application relates to the technical field of traditional Chinese medicines, and particularly discloses application of a traditional Chinese medicine composition in preparation of a medicine for treating organ fibrosis. The traditional Chinese medicine composition is prepared from raw medicinal materials in the following proportions by weight: Semen Euryae 150-350, Fructus Lycii 100-200, Schisandra chinensis 30-60, Cnidium 20-50, Rosa laevigata 20-50, Semen Allii Tuberosi 20-50, Dipsacus asper 20-50, Cistanche 20-50, Rehmannia glutinosa 30-60, Cyathula 20-50, Herba Epimedii 40-100, Rubus chingii 20-50, Ginseng 15-35, Cornu Cervi 10-25, sea horse 15-35, Fructus Meliae Toosendan 15-35. The traditional Chinese medicine composition can be applied in preparation of the medicine for treating organ fibrosis, the obtained medicine has the effects of inhibiting lung fibrosis, reducing myocardial fibrosis and reducing kidney fibrosis, the effect is remarkable, the application is safe, and the price is low.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, and particularly relates to application of a traditional Chinese medicine composition in preparation of a medicine for treating organ fibrosis. BACKGROUND

[0002] Fibrosis is an important pathological change of various diseases, and its causes are complex, which can cause damage or even loss of normal tissue function. The main pathological change of fibrosis is the increase of fibrous connective tissue in organ tissue, and the decrease of parenchymal cells. Continuous progression can cause destruction of organ structure and reduction of function, and even failure. Fibrosis can be caused by chronic inflammatory reactions caused by infection, autoimmune reaction, allergic reaction and chemical radiation stimulation, and the common pathway is the activation and proliferation of fibroblasts. Fibrosis can occur in multiple organs, such as lung, liver, kidney, heart and the like, leading to organ fibrosis.

[0003] Organ fibrosis is a pathological change of the increase of fibrous connective tissue and the decrease of parenchymal cells in the tissue of various organs caused by various acute and chronic diseases. It is caused by excessive reconstruction, hardening and scar formation of various organ tissues, and the common pathological basis is the abnormal increase and excessive deposition of extracellular matrix (ECM). Among them, kidney fibrosis is the common pathway of various chronic kidney diseases progressing to end-stage renal failure, and its essence is the "scarring" repair after kidney tissue damage; pulmonary fibrosis is the final outcome of the development, evolution and scarring of lung diseases; liver fibrosis is a repair response of the liver to chronic injury, and is a common pathological change of chronic liver disease; myocardial fibrosis is an important marker of decompensated myocardial hypertrophy and heart failure, and participates in myocardial remodeling caused by hypertension, hypertrophic cardiomyopathy, heart failure and myocardial infarction.

[0004] At present, fibrosis of solid organs such as heart, liver, lung and kidney is the main cause of organ failure and patient disability and death. If the important organs such as heart, lung and liver are fibrotic and not treated in time and effectively, it will cause organ hardening or organ failure, harm human physical and mental health, and even endanger life. However, due to the lack of in-depth understanding of the pathogenesis of organ fibrosis, there is a lack of biomarkers for early diagnosis and early warning in clinical practice, and there are almost no treatment drugs specifically for fibrosis.

[0005] Although organ transplantation is the most effective clinical solution in the terminal stage of organ fibrosis, but due to the problems of donor shortage, low survival rate, high cost, risk of rejection and infection after transplantation and the like, most patients cannot benefit from it, therefore, it is still a clinical problem to be solved to find other treatment solutions with more significant effect, safer application and simpler operation. SUMMARY

[0006] In view of the above technical problems existing in the prior art of treating senile organ fibrosis, the application provides application of a traditional Chinese medicine composition in preparation of a medicine for treating organ fibrosis.

[0007] To achieve the above-mentioned application purposes, the application adopts the following technical solutions:

[0008] The application provides application of a traditional Chinese medicine composition in preparation of a medicine for treating organ fibrosis, and the traditional Chinese medicine composition is prepared from raw medicinal materials in the following proportions by weight: Semen Eurycoma 150-350, Fructus Lycii 100-200, Schisandra chinensis 30-60, Cnidium 20-50, Rosa laevigata 20-50, Semen Allii Tuberosi 20-50, Radix Morindae Officinalis 20-50, Cistanche 20-50, Rehmannia glutinosa 30-60, Cyathula 20-50, Herba Epimedii 40-100, Rubus chingii 20-50, Radix et Rhizoma Ginseng 15-35, Cornu Cervi Pantotrichum 10-25, Cornu Callis 15-35, Fructus Meliae Toosendan 15-35.

[0009] The eight kinds of seeds in the traditional Chinese medicine composition of the application have the functions of tonifying kidney and essence, and the Semen Eurycoma, Fructus Lycii, Schisandra chinensis and Rubus chingii have the functions of tonifying kidney and essence, the Cnidium and Semen Allii Tuberosi have the functions of promoting yang and essence, and the Fructus Meliae Toosendan is not included in the tonifying and benefiting category, but is a qi-regulating product, and in the qi-regulating drugs, the Fructus Meliae Toosendan is good at regulating the qi of the lower jiao, liver and kidney, and is combined with the drugs for tonifying kidney essence, and has the function of tonifying without stagnation.

[0010] Further, the traditional Chinese medicine composition is prepared from raw medicinal materials in the following proportions by weight: Semen Eurycoma 150, Fructus Lycii 200, Schisandra chinensis 30, Cnidium 50, Rosa laevigata 20, Semen Allii Tuberosi 20, Radix Morindae Officinalis 50, Cistanche 20, Rehmannia glutinosa 30, Cyathula 50, Herba Epimedii 40, Rubus chingii 50, Radix et Rhizoma Ginseng 15, Cornu Cervi Pantotrichum 25, Cornu Callis 15, Fructus Meliae Toosendan 35.

[0011] Further, the traditional Chinese medicine composition is prepared from raw medicinal materials in the following proportions by weight: Semen Eurycoma 350, Fructus Lycii 100, Schisandra chinensis 60, Cnidium 20, Rosa laevigata 50, Semen Allii Tuberosi 50, Radix Morindae Officinalis 20, Cistanche 50, Rehmannia glutinosa 60, Cyathula 20, Herba Epimedii 100, Rubus chingii 20, Radix et Rhizoma Ginseng 35, Cornu Cervi Pantotrichum 10, Cornu Callis 35, Fructus Meliae Toosendan 15.

[0012] Further, the traditional Chinese medicine composition is prepared from raw medicinal materials in the following proportions by weight: Semen Eurycoma 250, Fructus Lycii 138, Schisandra chinensis 46, Cnidium 35, Rosa laevigata 35, Semen Allii Tuberosi 35, Radix Morindae Officinalis 35, Cistanche 35, Rehmannia glutinosa 46, Cyathula 35, Herba Epimedii 70, Rubus chingii 35, Radix et Rhizoma Ginseng 25, Cornu Cervi Pantotrichum 16, Cornu Callis 21, Fructus Meliae Toosendan 23.

[0013] Further, the traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Semen Eurycoma 30, Radix Rehmanniae 45, Fructus Corni 150, Schisandra chinensis 45, Cnidium 30, Chrysanthemum 30, Rubus 30, Cistanche 30, Radix Paeoniae Alba 45, Radix Cyathulae 30, Herba Epimedii 70, Wolfberry 30, Ginseng 20, Deer Antler 19, Sea Horse 20, Fructus Meliae 20.

[0014] Further, the dosage form of the medicine is decoction, capsule, tablet, granule, powder or pill.

[0015] Further, the preparation method of the capsule comprises the following steps:

[0016] a. Weigh Cnidium, Semen Eurycoma, Schisandra chinensis and Herba Epimedii according to the proportion, add 6-10 times of 70% ethanol, reflux extract 1-3 times, each time for 1-3 hours, filter, combine the filtrate, recover ethanol under reduced pressure until there is no alcohol taste, and obtain alcohol extract;

[0017] b. Combine the medicinal material residues after alcohol extraction in step a with proportioned Fructus Corni, Radix Rehmanniae, Rubus, Chrysanthemum, Semen Allii Tuberosi, Radix Morindae Officinalis and Fructus Meliae, add 7-12 times of water, decoct 1-3 times, each time for 1-3 hours, filter, combine the filtrate, mix with the alcohol extract obtained in step a, and concentrate under reduced pressure until the relative density is 1.25-1.30 at 60°C, and obtain extract;

[0018] c. Weigh Ginseng, Deer Antler, Sea Horse, Cistanche and Radix Cyathulae according to the proportion, crush into fine powder, mix with the extract obtained in step b, dry at 60-70°C, crush, sieve and fill into capsules.

[0019] Further, the organ fibrosis includes at least one of myocardial fibrosis, liver fibrosis, pulmonary fibrosis or kidney fibrosis.

[0020] The application also provides the use of the medicine composition in reducing liver inflammation.

[0021] The traditional Chinese medicine composition provided by the application can be applied to prepare a medicine for treating organ fibrosis including at least one of myocardial fibrosis, liver fibrosis, pulmonary fibrosis or kidney fibrosis, and reduce the degree of organ fibrosis, which is safe, effective and low in price. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 is the Masson staining diagram of lung tissue in the test example of the application;

[0023] Figure 2 is the Masson staining diagram of heart tissue under different magnifications in the test example of the application;

[0024] Figure 3is an immunofluorescence staining chart in the test example of the present application;

[0025] Figure 4 is a kidney tissue staining chart in the test example of the present application;

[0026] Figure 5 is a liver HE staining chart in the test example of the present application;

[0027] Figure 6 is a liver Masson staining chart in the test example of the present application;

[0028] Figure 7 HAI (Hepatic Activity Index) is a liver histological activity index. DETAILED DESCRIPTION

[0029] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below in conjunction with examples. It should be understood that the specific examples described herein are only used to explain the present application and are not used to limit the present application.

[0030] Example 1: Preparation of capsules

[0031] Composition: Cuscuta 250g, Medlar 138g, Schisandra 46g, Cnidium 35g, Cherryl 35g, Chinese chive seed 35g, Dipsacus 35g, Cistanche 35g, Rehmannia 46g, Cyathula 35g, Herba Epimedii 70g, Raspberry 35g, Ginseng 25g, Deerhorn 16g, Sea-horse 21g, Melia 23g.

[0032] Preparation method:

[0033] a. Cnidium, Cuscuta, Schisandra, Herba Epimedii were weighed according to the proportion, 3208ml of 70% ethanol was added, and reflux extraction was carried out for 3 times, 2 hours each time, and the filtrate was combined. The ethanol was recovered under reduced pressure until there was no alcohol taste, and the alcohol extract was obtained;

[0034] b. The medicinal material residues after alcohol extraction in step a were combined with proportioned Medlar, Rehmannia, Raspberry, Cherryl, Chinese chive seed, Dipsacus, and Melia, and 6732ml of water was added for decoction for 2 times, 2 hours each time. The filtrate was combined and mixed with the alcohol extract obtained in step a. The mixture was concentrated under reduced pressure until the relative density was 1.28 at 60°C, and the extract was obtained;

[0035] c. Ginseng, Deerhorn, Sea-horse, Cistanche, and Cyathula were weighed according to the proportion, crushed into fine powder, and mixed with the extract obtained in step b. The mixture was dried at 65°C, crushed, sieved, filled into capsules, and 757 capsules were obtained.

[0036] Example 2: Preparation of tablets

[0037] Ingredients: Cuscuta 250g, Medlar 150g, Schisandra 45g, Cnidium 30g, Cherryl 30g, Chinese chive seed 30g, Ba Ji Tian 30g, Cistanche 30g, Rehmannia 45g, Cow's hoof 30g, Herba Epimedii 70g, Raspberry 30g, Ginseng 20g, Deer horn 19g, Sea horse 20g, Melia 20g.

[0038] Preparation method:

[0039] a. Cuscuta, Cnidium, Schisandra, Herba Epimedii were weighed according to the proportion, 2765ml 70% ethanol was added, reflux extraction was carried out for 3 times, 1 hour each time, filtration was carried out, the filtrate was combined, ethanol was recovered under reduced pressure until there was no alcohol taste, and alcohol extraction extract was obtained;

[0040] b. The medicinal material residue after alcohol extraction in step a was combined with proportioned Medlar, Rehmannia, Raspberry, Cherryl, Chinese chive seed, Ba Ji Tian, and Melia, 5110ml water was added for decoction for 1 time, 3 hours, filtration was carried out, the filtrate was combined, mixed with the alcohol extraction extract obtained in step a, and concentrated under reduced pressure until the relative density was 1.30 at 60℃, and the extract was obtained;

[0041] c. Ginseng, Deer horn, Sea horse, Cistanche, and Cow's hoof were powdered and mixed with the extract obtained in step b, dried at 70℃, powdered, sieved, 1% magnesium stearate was added, and tableting was carried out, and 760 tablets were obtained.

[0042] Example 3: Preparation of pills

[0043] Ingredients: Cuscuta 350g, Medlar 100g, Schisandra 60g, Cnidium 20g, Cherryl 50g, Chinese chive seed 50g, Ba Ji Tian 20g, Cistanche 50g, Rehmannia 60g, Cow's hoof 20g, Herba Epimedii 100g, Raspberry 20g, Ginseng 35g, Deer horn 10g, Sea horse 35g, Melia 15g.

[0044] Preparation method:

[0045] a. Cuscuta, Cnidium, Schisandra, Herba Epimedii were weighed according to the proportion, 2765ml 70% ethanol was added, reflux extraction was carried out for 3 times, 1 hour each time, filtration was carried out, the filtrate was combined, ethanol was recovered under reduced pressure until there was no alcohol taste, and alcohol extraction extract was obtained;

[0046] b. The medicinal material residue after alcohol extraction in step a was combined with proportioned Medlar, Rehmannia, Raspberry, Cherryl, Chinese chive seed, Ba Ji Tian, and Melia, 5110ml water was added for decoction for 1 time, 3 hours, filtration was carried out, the filtrate was combined, mixed with the alcohol extraction extract obtained in step a, and concentrated under reduced pressure until the relative density was 1.30 at 60℃, and the extract was obtained;

[0047] c. Weigh the ginseng, deer horn, sea horse, cistanche, and Sichuan cow's hoof according to the proportion, crush into fine powder, mix with the extract obtained in step b, dry at 60°C, crush, sieve, and make into pills to obtain 320 g of pills.

[0048] Example 4: Preparation of granules

[0049] Composition: Cuscuta 150 g, medlar 200 g, Schisandra 30 g, Cnidium 50 g, cherry 20 g, Chinese chive seed 20 g, Ba Ji Tian 50 g, Cistanche 20 g, Rehmannia 30 g, Sichuan cow's hoof 50 g, Herba Epimedii 40 g, raspberry 50 g, ginseng 15 g, deer horn 25 g, sea horse 15 g, Sichuan orange fruit 35 g.

[0050] Preparation method:

[0051] a. Weigh Cnidium, Cuscuta, Schisandra, and Herba Epimedii according to the proportion, add 2430 ml of 70% ethanol, reflux extract 2 times, 1.5 hours each time, filter, combine the filtrate, and recover ethanol under reduced pressure until there is no alcohol taste to obtain alcohol extract;

[0052] b. Combine the residue of the medicinal materials after alcohol extraction in step a with the proportion of the remaining medicinal materials, add 8800 ml of water, decoct 3 times, 1 hour each time, filter, combine the filtrate, mix with the alcohol extract obtained in step a, and concentrate under reduced pressure to 1.29 in relative density at 60°C to obtain extract;

[0053] c. Mix the extract obtained in step b with 300 g of dextrin and 200 g of sucrose powder uniformly, dry at 70°C, crush, sieve, and granulate to obtain 700 g of granules.

[0054] Example 5: Preparation of powder

[0055] Composition: Cuscuta 250 g, medlar 150 g, Schisandra 45 g, Cnidium 30 g, cherry 30 g, Chinese chive seed 30 g, Ba Ji Tian 30 g, Cistanche 30 g, Rehmannia 45 g, Sichuan cow's hoof 30 g, Herba Epimedii 70 g, raspberry 30 g, ginseng 20 g, deer horn 19 g, sea horse 20 g, Sichuan orange fruit 20 g.

[0056] Preparation method: Weigh the medicinal materials in the composition according to the proportion, mix and crush into fine powder to obtain 823 g of powder.

[0057] Example 6: Preparation of decoction

[0058] Composition: Cuscuta 250 g, medlar 150 g, Schisandra 45 g, Cnidium 30 g, cherry 30 g, Chinese chive seed 30 g, Ba Ji Tian 30 g, Cistanche 30 g, Rehmannia 45 g, Sichuan cow's hoof 30 g, Herba Epimedii 70 g, raspberry 30 g, ginseng 20 g, deer horn 19 g, sea horse 20 g, Sichuan orange fruit 20 g.

[0059] Preparation method: Weigh the Chinese medicinal materials according to the proportion, add water to cover the medicinal materials, soak for 15 minutes, heat to boiling, continue to simmer for 30 minutes, filter, add water to the dregs again to cover the medicinal materials, simmer for another 30 minutes, filter, combine the filtrates to obtain 4320g of decoction.

[0060] Experimental example:

[0061] To clarify the effect of the drug of the present invention on improving the degree of organ fibrosis and weakness, the following experiments were conducted using Example 1 (hereinafter referred to as the drug of the present invention or BZBS):

[0062] 1. Animal experiments on pulmonary fibrosis

[0063] A mouse pulmonary fibrosis model was established using a single intratracheal injection of bleomycin. Sixty-four male ICR mice (SPF grade), weighing 22-25g, were randomly divided into a control group, a model group, a pirfenidone (manufacturer: Beijing Kangtini Pharmaceutical Co., Ltd.) group, and a BZBS group. Animals underwent intratracheal injection; the control group received saline, while the other groups received bleomycin (manufacturer: Nippon Kayaku Co., Ltd.). The BZBS group received a dose equivalent to 2g crude drug / kg / day, and the pirfenidone group received a dose of 300mg / kg / day. Animals in the control and model groups were administered the corresponding volumetric solvent (CMC) via gavage. Administration began on the day of model establishment and continued for 21 days.

[0064] After drug administration, the animals were weighed to observe changes in body weight. The entire lung was weighed and then placed in liquid nitrogen to calculate the lung coefficient. The results are shown in Table 1. The lung tissue hydroxyproline, a fibrosis marker, was tested, and the results are shown in Table 2. Some animals underwent lung perfusion fixation and Masson staining to detect lung fibrosis. The results are shown in Tables 3 and 4. Figure 1 (Masson staining images of lung tissue (20×), including the blank group, model group, pirfenidone group and BZBS group) are shown.

[0065]

[0066] From the above data, the model group compared with the blank group, the model group lung coefficient was significantly increased (P<0.01), and compared with the model group, the BZBS group and the pirfenidone group were significantly reduced (P<0.05); At the same time, the model group compared with the blank group, the model group hydroxyproline content was significantly increased (P<0.05); Compared with the model group, the BZBS group and the pirfenidone group were significantly reduced (P<0.05); In addition, compared with the model group, the BZBS group and the pirfenidone group had relatively low pulmonary fibrosis score. It is proved that BZBS can inhibit the degree of bleomycin-induced pulmonary fibrosis in mice, and has the effect of preventing and treating pulmonary fibrosis.

[0067] 2. Myocardial fibrosis animal experiment

[0068] Male SD rats (SPF level) 50, all 2 months old, animal weight 200±10g, purchased from Beijing Vito Lihua Experimental Animal Technology Co., Ltd. The animal feeding conditions are 12 hours of light per day, temperature 20~26℃, relative humidity 40~70%, fed with maintenance feed (provided by Beijing Kaoshe Lihua Feed Co., Ltd.), and drank clean water prepared by ROA50 experimental animal water system. The animals were quarantined for 3 days and adaptively fed for 2 days. During this period, the animal's drinking, eating and health status, as well as the presence of disease and death signs were observed.

[0069] The 50 SD rats were randomly divided into 5 groups by random number table method: normal group (normal diet+ equal volume of solvent gavage), model group (normal diet+ equal volume of solvent gavage), BZBS low dose group (normal diet+ BZBS 1g / kg / d gavage), BZBS high dose group (normal diet+ BZBS 4g / kg / d gavage), sacubitril valsartan control group (normal diet+ 20mg / kg / 2d gavage), and drug intervention was given (as shown in Table 4). The test product was given as follows: the BZBS high dose group, the BZBS low dose group and the control group were given 10mL / kg of drug solution by gavage every day, and the normal group and the model group were given equal volume of solvent CMC by gavage every day.

[0070]

[0071] The model making method is as follows: the SD rats are adaptively fed for 1 week, weighed, and then injected with 10% chloral hydrate (0.35 g / kg) intraperitoneally; after anesthesia, the right lateral position is fixed on the mouse plate, the right costal region is shaved, and the left kidney region is exposed; 75% alcohol is used for disinfection in the kidney region, a longitudinal incision is made on the left side of the twelfth costal margin of the spine, about 1-2 cm long, the muscle layer is bluntly dissected, the kidney is found and exposed after extrusion, the fat around the kidney is dissected, and the renal artery and vein and ureter are exposed; the kidney is fixed with a hemostat and the entire artery and vein and ureter are ligated with 4-0 surgical suture, and the kidney is cut off; the splanchnic layer and muscle layer are successively sutured, 10,000 u of penicillin is injected intraperitoneally, and the excess blood is wiped off; the rat is placed on an electric heating blanket for heat preservation, and after the vital signs are stable, the rat is returned to the mouse cage for feeding; 1% saline is used for drinking water, and the rat is fed freely. After 4 weeks, the rat heart function is detected using a small animal ultrasound instrument, and EF>50% and a reduced E / A ratio are regarded as successful establishment of a chronic heart failure model.

[0072] After 2 months of administration, the rats are anesthetized with isoflurane, and the rats are waited until they lose consciousness. After the rats are anesthetized, blood is taken from the abdominal aorta, and after standing at room temperature for 2 h, the serum is taken after centrifugation at 3,000 r for 15 min, and is stored at -80°C for standby use. The heart tissue is placed in a 4% paraformaldehyde solution for fixation, and a part is placed in a cryopreservation tube and stored at -80°C for standby use. Part of the heart is sent for Masson and HE staining, and the morphological changes of the heart tissue are observed. The results are shown in Figure 2 As shown in the table, the Masson staining of the normal group, the model group, the control group, the BZBS low-dose group and the BZBS high-dose group under different magnifications (10x, 20x and 40x) is shown. The Masson staining shows that the myocardial cells of the normal group are arranged in order, the cells are not degenerated and necrotic, and there is no obvious collagen fiber proliferation. A large number of muscle fiber rupture, atrophy, degeneration, and collagen fiber blue staining are observed in the model group, the degree of myocardial fibrosis is significantly increased, and the myocardial cells are stiff. The myocardial fibrosis degree of each administration group (control group, BZBS low-dose group and BZBS high-dose group) is reduced to different degrees.

[0073] At the same time, the myocardial cell cross-sectional area is measured by immunofluorescence. The myocardial tissue section is deparaffinized, the antigen is repaired, the endogenous peroxidase is blocked, 3% BSA is blocked at room temperature for 30 min, the primary antibody is incubated at 4°C overnight, the secondary antibody is incubated for 1 h in the dark, DAPI (4', 6-diamidino-2-phenylindole) is used to stain the cell nucleus, and neutral resin is used for mounting. Three discontinuous fields of view are randomly selected under a microscope, and the staining results are observed, as shown in Figure 3As shown in the immunofluorescence staining images (20x), wheat germ agglutinin (WGA) appears green, a-smooth muscle actin (a-SMA) appears red, and DAPI appears blue. The brown-yellow particles are positive cells. WGA staining shows that the cross-sectional area of myocardial cells in the model group is larger than that in the normal group, which morphologically confirms the presence of myocardial hypertrophy. Each administration group (control group, BZBS low-dose group, and BZBS high-dose group) can significantly restore the size of myocardial cells (P<0.05). There is no significant difference in the expression of a-SMA around blood vessels in each administration group.

[0074] From the above data, it can be seen that BZBS can significantly reduce the collagen content in the myocardial tissue of chronic heart failure rats, reduce the cross-sectional area of myocardial cells, reduce the degree of myocardial fibrosis, inhibit left ventricular remodeling, and thus improve the cardiac function of rats.

[0075] 3. Kidney fibrosis animal experiment

[0076] Fifteen male BKS-DB (WT) mice (body weight about 19-30 g) were used as a blank group, and 45 male BKS-DB mice (body weight about 40-69 g) were used as an experimental group. The animals were obtained from Jiangsu Jizhu Pharmaceutical Biotechnology Co., Ltd. The experimental group animals were given a high-fat and high-protein diet at 8 weeks of age, and after continuous feeding for 8 weeks, the blood glucose and urine biochemistry (mAlb / CREA) indicators of the animals were detected to reach the symptoms of diabetic nephropathy. The experimental group animals were divided into a model group, a BZBS group, and a dapagliflozin group according to blood glucose, with 15 animals in each group. The animals were given drugs on the day of grouping, once a day, and continuously for 6 weeks. The BZBS group was given a dose of 1.44 g of crude drug / kg / d, and the dapagliflozin group was given a dose of 1 mg / kg / d. The blank group and the model group were given an equal volume of water. After the administration was completed, the kidney fibrosis and urine biochemistry of each group were detected, and the results are shown in Tables 5 and 6. Figure 4 (renal tissue staining images (10x)) and Tables 5 and 6. Compared with the model group, the urine biochemistry indicators of the BZBS group were significantly reduced (P<0.05).

[0077]

[0078] From the above data, it can be seen that after 6 weeks of BZBS administration, the urine biochemistry (mAlb / CREA) indicators of the test animals were significantly improved, and the degree of kidney fibrosis was significantly reduced.

[0079] 4. Liver fibrosis animal experiment

[0080] Test purposes

[0081] The regulation of the drug composition of the present application on the inflammation and fibrosis of the liver of naturally aging mice was studied.

[0082] 1 Experimental materials

[0083] 1.1 Test sample

[0084] 1.1.1 Name: pharmaceutical composition of the present application (BZBS).

[0085] 1.1.2 Route and dosage: oral administration

[0086] 1.1.3 Source and batch number: Shijiazhuang Yiling Pharmaceutical Co., Ltd., batch number XB2103001.

[0087] 1.2 Tool drugs and main reagents and materials

[0088] Sodium carboxymethylcellulose (CMC-Na): Tianjin Damao Chemical Reagent Factory, batch number: 20181006, expiration date: December 30, 2021.

[0089] 1.3 Experimental system

[0090] 1.3.1 Animal strain: C57BL / 6J mice.

[0091] 1.3.2 Animal level: SPF level.

[0092] 1.3.3 Animal gender and quantity: 60 male mice were purchased, and 30 of them (30 / male) were selected for this test. In addition, 12 mice of the same age, gender, and manufacturer were directly used as controls.

[0093] 1.3.4 Animal age at the start of administration: 52 weeks old.

[0094] 1.3.5 Animal body weight at the start of administration: actual 20-40g.

[0095] 1.3.6 Animal source: purchased from Beijing Vantolliva Experimental Animal Technology Co., Ltd.

[0096] 1.3.7 Animal qualification certificate number, issuing unit, and receiving date: qualification certificate number 1100112011043405, license number SCXK (Jing) 2016-0006, issued by Beijing Vantolliva Experimental Animal Technology Co., Ltd., received on July 15, 2020.

[0097] 1.3.7 Rearing conditions: mice were kept in cages and reared in the New Drug Evaluation Center of Hebei Yiling Pharmaceutical Research Institute Co., Ltd., with 12 hours of light per day, temperature 20-26℃, relative humidity 40-70%, and experimental animal use license number: SYXK (Ji) 2020-003.

[0098] 1.3.8 Quarantine process: the animals are quarantined for 3 days, and adaptively fed for 2 days, during which the animals' drinking, eating and health conditions, and the presence of disease and death signs are observed.

[0099] 1.3.9 Feed: SPF size mouse maintenance feed is provided by Beijing Keao Huihe Feed Co., Ltd. The feed production license number is Jing Shi Zheng (2018) 06073, and the feed batch number is 20103213, 20113213.

[0100] 1.3.10 Drinking water: clean water prepared by ROA50 experimental animal drinking water system (instrument number YL-LE-A01) is drunk, and a filled drinking water bottle is provided for free drinking, and the water quality meets the "Drinking Water Health Standards".

[0101] 1.3.11 Bedding: ordinary corn cob bedding, which is used after high-temperature and high-pressure disinfection, is provided by Beijing Keao Huihe Feed Co., Ltd., and the bedding batch number is 20079811.

[0102] 1.3.12 Identification: experimental animal ear tag marking.

[0103] 2 Test method

[0104] 2.1 Test design basis

[0105] 2.1.1 Design basis: relevant literature.

[0106] 2.1.2 Experimental system selection explanation:

[0107] Old C57BL / 6J mice: research applications include immunology, cancer, longevity intervention, and biomarker research. The balance and stability of stem cells in 13-month-old C57 mice are disrupted, and the animals begin to age rapidly, so natural aging C57BL / 6J mice are used to simulate human premature aging.

[0108] 2.2 Dose and grouping

[0109] Divided into model group (equal volume of distilled water), drug composition group of the application (2g / kg / d), 23-week-old mice as young group, without any treatment, directly for sampling.

[0110] The daily dose of the drug composition of the application for humans is 2.4g of crude drug (human weight is 60kg), and the BZBS group of mice is defined as 2g of crude drug / kg. 30 C57 mice are randomly divided into 2 groups according to weight, including model group and drug composition group of the application.

[0111] 2.3 Test product administration method

[0112] Gavage, consistent with the intended route.

[0113] 2.4 Preparation and storage of test articles

[0114] The pharmaceutical composition of the present application (BZBS): 1.2 g of the pharmaceutical composition of the present application super-micro powder + 12 ml of CMC, prepared on the same day and used immediately

[0115] According to the dosage of 10 ml / kg and the dose setting, the preparation concentration was calculated (see Table 7), and the pharmaceutical composition of the present application was weighed according to the number of animals, ground and suspended with the solvent, made up to volume, and the test article was prepared and used immediately. The solvent was 0.5% CMC-Na, which was uniformly spread on the surface of pure water to swell fully, made up to volume, and stored at 2-8°C for use.

[0116]

[0117] 2.5 Administration of test articles

[0118] The test article was administered with a syringe with a volume of 10 ml / kg, and the model group and the BZBS group of animals were administered with the corresponding volume of solvent, which was replaced every 48 h.

[0119] 2.6 Model preparation: C57 mice were naturally bred to 13 months of age, which were natural aging animals.

[0120] 2.7 Detection index: histopathological analysis

[0121] Liver samples were fixed with 4% paraformaldehyde, dehydrated and embedded in paraffin. 5 μm thick sections were prepared, stained with hematoxylin-eosin (HE) and Masson, and histological analysis was performed under a light microscope. On the basis of the widely accepted histological activity index (HAI) (Knodell et al., 1981), we slightly modified the scoring criteria. In short, HAI represents the sum of the scores of necro-inflammatory lesions, i.e. periporal and bridging necrosis (0-10), degeneration and focal necrosis within lobules (0-4), inflammation (0-4), fibrosis (0-4). The liver-stained sections of each group were scored by two pathologists according to the HAI scoring criteria.

[0122] 3 Pathological results

[0123] 3.1 Results as shown in Table 3 Figure 5 Liver HE staining, central vein area, part of the periportal inflammation was observed in the natural aging model group, a small amount of inflammation was observed in the pharmaceutical composition of the present application group, and no inflammation was observed in the young group.

[0124] 3.2 Results as shown in Table 4 Figure 6 Liver masson staining, part of the fibrosis was observed in the natural aging model group, no fibrosis was observed in the pharmaceutical composition of the present application group and the young group.

[0125] 3.3 Results As shown in Figure 7 HAI (Hepatic Activity Index) The model group was 10 points, the drug composition group was 3 points, and the young group was 0 points. HAI index (n=10)

[0126] 4 Conclusion: The drug composition can reduce inflammation and fibrosis of the liver during the aging process.

[0127] In summary, the BZBS provided by the embodiments of the present application can inhibit the degree of pulmonary fibrosis caused by bleomycin in mice, reduce the cross-sectional area of myocardial cells of chronic heart failure rats, and reduce the degree of myocardial fibrosis. In addition, the BZBS can reduce the degree of kidney fibrosis in mice with symptoms of diabetic nephropathy, reduce inflammation and fibrosis of the liver during the aging process, and is used for treating organ fibrosis including myocardial fibrosis, pulmonary fibrosis, kidney fibrosis, and liver fibrosis. The drugs provided by embodiments 2 to 6 have substantially equivalent effects to the capsules in embodiment 1.

[0128] The above merely describes preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a medicine for treating organ fibrosis, characterized in that, The traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Cuscuta chinensis 150-350, Lycium barbarum 100-200, Schisandra chinensis 30-60, Cnidium monnieri 20-50, Rosa laevigata 20-50, Allium macranthum 20-50, Morinda officinalis 20-50, Cynomorium songaricum 20-50, Rehmannia glutinosa 30-60, Cyathula officinalis 20-50, Herba Epimedii 40-100, Rubus chingii 20-50, Panax ginseng 15-35, Cornu Cervi Pantotrichum 10-25, Dendrochirus 15-35, Melia toosendan 15-35.

2. Use according to claim 1, wherein The traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Cuscuta chinensis 150, Lycium barbarum 200, Schisandra chinensis 30, Cnidium monnieri 50, Rosa laevigata 20, Allium macranthum 20, Morinda officinalis 50, Rehmannia glutinosa 30, Cyathula officinalis 50, Herba Epimedii 40, Rubus chingii 50, Panax ginseng 15, Cornu Cervi Pantotrichum 25, Dendrochirus 15, Melia toosendan 35.

3. The use according to claim 1, wherein The traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Cuscuta chinensis 350, Lycium barbarum 100, Schisandra chinensis 60, Cnidium monnieri 20, Rosa laevigata 50, Allium macranthum 50, Morinda officinalis 20, Rehmannia glutinosa 60, Cyathula officinalis 20, Herba Epimedii 100, Rubus chingii 20, Panax ginseng 35, Cornu Cervi Pantotrichum 10, Dendrochirus 35, Melia toosendan 15.

4. The use according to claim 1, wherein The traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Cuscuta chinensis 250, Lycium barbarum 138, Schisandra chinensis 46, Cnidium monnieri 35, Rosa laevigata 35, Allium macranthum 35, Morinda officinalis 35, Rehmannia glutinosa 46, Cyathula officinalis 35, Herba Epimedii 70, Rubus chingii 35, Panax ginseng 25, Cornu Cervi Pantotrichum 16, Dendrochirus 21, Melia toosendan 23.

5. The use according to claim 1, wherein the compound is ###0002### The traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: Cuscuta chinensis 250, Lycium barbarum 150, Schisandra chinensis 45, Cnidium monnieri 30, Rosa laevigata 30, Allium macranthum 30, Morinda officinalis 30, Rehmannia glutinosa 45, Cyathula officinalis 30, Herba Epimedii 70, Rubus chingii 30, Panax ginseng 20, Cornu Cervi Pantotrichum 19, Dendrochirus 20, Melia toosendan 20.

6. Use according to any one of claims 1 to 5, characterized in that, The dosage form of the medicine is decoction, capsule, tablet, granule, powder or pill.

7. Use according to claim 6, wherein The preparation method of the capsule comprises the following steps: a. Cnidium monnieri, Cuscuta chinensis, Schisandra chinensis and Herba Epimedii are weighed in proportion, 6-10 times the amount of 70% ethanol is added, and reflux extraction is performed 1-3 times, each time for 1-3 hours, the filtrate is combined, and the ethanol is recovered under reduced pressure until no alcohol taste is left, to obtain alcohol extraction extract; b. the medicinal material residues after alcohol extraction in step a are combined with proportionally weighed Lycium barbarum, Rehmannia glutinosa, Rubus chingii, Rosa laevigata, Allium macranthum, Morinda officinalis and Melia toosendan, 7-12 times the amount of water is added, decoction is performed 1-3 times, each time for 1-3 hours, the filtrate is combined, and mixed with the alcohol extraction extract obtained in step a, concentrated under reduced pressure to a relative density of 1.25-1.30 at 60°C, to obtain extract; c. Panax ginseng, Cornu Cervi Pantotrichum, Dendrochirus, Cynomorium songaricum and Cyathula officinalis are weighed in proportion, crushed into fine powder, mixed with the extract obtained in step b, dried at 60-70°C, crushed, sieved and filled into capsules.

8. Use according to any one of claims 1 to 5, wherein The organ fibrosis includes at least one of myocardial fibrosis, liver fibrosis, pulmonary fibrosis or renal fibrosis.

9. Use according to any one of claims 1 to 5, wherein The traditional Chinese medicine composition can reduce liver inflammation while treating organ fibrosis.

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