CN115212253A - Chinese herbal compound preparation for treating hepatic fibrosis and preparation method and application thereof - Google Patents
Chinese herbal compound preparation for treating hepatic fibrosis and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a compound traditional Chinese medicine preparation for treating hepatic fibrosis, which is prepared from fiveleaf akebia fruit, polygonum, pennycress and Chinese holly fruit according to a certain weight proportion by mixing, and the preparation method comprises the steps of adding 10 times of 60% ethanol into the raw materials, heating and refluxing for 3 times, and extracting for 2 hours each time. Mixing the extractive solutions, recovering ethanol, concentrating, evaporating, drying under reduced pressure, adding appropriate amount of adjuvants corresponding to different dosage forms, and making into tablet, capsule, granule, pill, etc. by conventional preparation method. Pharmacodynamic test results show that the Chinese medicinal compound preparation has the effect of remarkably improving hepatic fibrosis lesion and injury.
Description
Technical Field
The invention relates to a Chinese medicinal compound preparation for treating hepatic fibrosis and a preparation method and application thereof, belonging to the field of Chinese medicinal pharmacy.
Background
Hepatic fibrosis is a compensatory reaction in the process of tissue repair after various chronic liver injuries, and reversing hepatic fibrosis can block the evolution of chronic liver diseases to cirrhosis and even liver cancer. During hepatic fibrosis, a large amount of fibrous nodules and pseudolobules are formed in the liver, so that the structure of the liver is damaged, the function of the liver is abnormal, the resistance of hepatic sinuses is increased, and the liver tissue is pathologically overloaded with iron, so that extracellular matrix (ECM) accumulation, hepatocyte necrosis, hepatic Stellate Cell (HSC) activation and the like caused by the hepatic fibrosis are not only the pathogenic essence of hepatic fibrosis, but also important synergistic pathogenic factors for further promoting the development of liver cirrhosis and liver cancer. Numerous basic and clinical studies have demonstrated that fibrotic liver ECM is produced primarily by secretion of activated HSCs. When the liver is in normal physiological conditions, HSCs appear to be in a quiescent state, star-shaped in appearance, lipid droplet-rich intracellular, high expression of Glial Fibrillary Acidic Protein (GFAP), low expression of alpha-smooth actin (alpha-SMA), and no or only a small amount of ECM synthesis; when the liver is stimulated by the injury factor, the HSC is in an activated state, the appearance of the HSC presents a fusiform shape, a large amount of intracellular lipid droplets are lost, alpha-SMA is highly expressed, GFAP is lowly expressed, a large amount of synthesized ECM is deposited on Dou Jianxi, the exchange of materials and information between the hepatic sinus and the hepatic cell is blocked, and the occurrence and the development of hepatic fibrosis are induced. Unfortunately, at present, there is no hepatic fibrosis treatment strategy with exact curative effect in clinic, and how to effectively prevent and treat hepatic fibrosis is a great problem which puzzles the medical community at present. The deep research on the potential pathological mechanism and the intervention target of the hepatic fibrosis and the research and development of safe and effective prevention and treatment medicines based on the deep research have great clinical significance. Under the guidance of the theories of 'treatment based on syndrome differentiation' and 'holism', the traditional Chinese medicine has unique cognition on prevention and treatment of hepatic fibrosis through long-term practice. The traditional Chinese medicine theory considers that the pathogenesis essence of hepatic fibrosis is liver channel obstruction, blood stasis and qi stagnation, so the basic principle of activating blood circulation to dissipate blood stasis and promoting qi circulation to remove meridian obstruction is used in treatment. The traditional Chinese medicine prescription is closely combined with clinical symptoms, and can fully exert the comprehensive action advantages of multiple components, multiple ways and multiple targets.
In order to solve the problems, the invention aims to provide a traditional Chinese medicine compound for treating hepatic fibrosis and a preparation method thereof.
Disclosure of Invention
The invention aims to provide a Chinese herbal medicine compound preparation for treating hepatic fibrosis, which has the effects of improving the pathological change of hepatic fibrosis and reducing collagen deposition. The invention also aims to provide a preparation method of the traditional Chinese medicine compound preparation.
A traditional Chinese medicine compound preparation for treating hepatic fibrosis is prepared from the following raw material medicines in parts by weight: 20-40 parts of fiveleaf akebia fruit, 10-30 parts of Polygonum criopolitanum, 5-15 parts of Thlaspis arvense and 3-8 parts of Chinese holly fruit.
The traditional Chinese medicine compound preparation for treating hepatic fibrosis is prepared from the following raw material medicines in parts by weight: 20 parts of fiveleaf akebia fruit, 30 parts of Polygonum multiflorum fruit, 5 parts of Thlaspi arvense and 8 parts of Chinese holly fruit.
The traditional Chinese medicine compound preparation for treating hepatic fibrosis is prepared from the following raw material medicines in parts by weight: 40 parts of fiveleaf akebia fruit, 10 parts of Polygonum fruit, 15 parts of Thlaspi arvense and 3 parts of Chinese holly fruit
The traditional Chinese medicine compound preparation for treating hepatic fibrosis is prepared from the following raw material medicines in parts by weight: 30 parts of fiveleaf akebia fruit, 20 parts of Polygonum flaccidum fruit, 10 parts of Thlaspis arvense seed and 5 parts of Chinese holly fruit.
The preparation method of the traditional Chinese medicine compound preparation for treating hepatic fibrosis is as follows: taking fructus Akebiae, fructus Polygoni Caespitosi, semen Thlaspis, and fructus Ilicis Cornutae, adding 10 times of 60% ethanol, heating and reflux-extracting for 3 times, each for 2 hr, mixing extractive solutions, recovering ethanol, adding appropriate adjuvant, and making into oral liquid by conventional preparation method.
The preparation method of the traditional Chinese medicine compound preparation for treating hepatic fibrosis is as follows: taking fructus Akebiae, fructus Polygoni Caespitosi, semen Thlaspis, and fructus Ilicis Cornutae, adding 10 times of 60% ethanol, heating and reflux-extracting for 3 times, each for 2 hr, mixing extractive solutions, recovering ethanol, concentrating, evaporating, drying under reduced pressure, pulverizing into fine powder, adding appropriate adjuvant, and making into granule, tablet, capsule or pellet by conventional preparation method.
The preparation method of the traditional Chinese medicine compound preparation for treating hepatic fibrosis adopts the condition of reduced pressure drying at the temperature of 60-70 ℃ and the pressure of 0.08mpa.
The Chinese herbal compound preparation is applied to the preparation of the medicine for treating hepatic fibrosis diseases.
The traditional Chinese medicine compound preparation is derived from a scientific research formula of Nanjing Chinese medicine university, is obtained by screening four traditional Chinese herbal medicines of fiveleaf akebia fruit, polygonum fruit, thlaspis and Chinese holly fruit through strict standards, has the characteristics of compatibility of medicines, overall regulation, treatment of both principal and secondary aspect of disease and the like, and has obvious curative effect on hepatic fibrosis diseases. The traditional Chinese medicine pathogenesis of hepatic fibrosis is 'blood stasis blockage' as the main pathogenesis, the stagnation of qi leads to unsmooth blood circulation and blood stasis obstruction, and the function of liver governing blood collection also determines that the liver blood stasis is easy to cause under the pathological state. Liver qi regulates qi of the whole body and is the viscera of "storing blood", so qi movement is obstructed and blood stasis occurs in the liver first, resulting in liver fibrosis. Based on the characteristics of the disease, the invention applies a clinical method of activating blood circulation to dissipate blood stasis, eliminating dampness and strengthening spleen, and breaking symptoms and removing stagnation to prepare the traditional Chinese medicine compound preparation for treating hepatic fibrosis. On the basis of the medicines for soothing the liver and activating blood circulation, the invention adds the products for removing blood stasis and dissipating stagnation, nourishing and clearing heat, and clearing liver and improving vision, and can also focus on treating the disease while strengthening the body resistance and eliminating the pathogenic factor, and focus on regulating and nourishing the viscera.
Monarch drug: fiveleaf akebia fruit, a mature fruit of Akebla quinata of the family Lardizabalaceae. The book of Sichuan Chinese materia medica records that: its advantages are mild nature, bitter and pungent taste, and no poison. It enters liver, stomach and bladder meridians. Has effects in dispersing stagnated liver qi, regulating stomach function, promoting blood circulation, relieving pain, softening and resolving hard mass, and promoting urination, and can be used for treating stagnation of qi of liver and stomach, goiter, lymphoid tuberculosis, and malignant tumor. Shanxi Chinese herbal medicine records that: to soothe the liver and tonify the kidney, strengthen the spleen and harmonize the stomach. The record of the Sichuan Chinese materia medica book is as follows: to soothe the liver and receive kidney qi. It is indicated for hernia and hematemesis. The clinical utility of anticancer traditional Chinese medicine is recorded as follows: it is used for treating goiter and scrofula caused by phlegm-fire consolidation. Modern researches show that fiveleaf akebia fruit mainly contains fatty oil, reducing sugar and various enzymes, and the extract of fiveleaf akebia fruit has the effects of resisting tumors and oxidative damage. Fructus akebiae is used as a monarch drug, has the effects of activating qi, relieving epigastric distention, dredging and removing stagnation, and mainly has the effects of soothing liver, regulating qi, removing blood stasis, resolving hard mass, removing blood stasis, stopping bleeding, promoting blood circulation and relieving pain, and can be used for treating various liver injuries and has a main treatment effect on hepatic fibrosis.
Ministerial drugs: polygonum hydropiper L.is fruit of Polygonum hydropiper L.of Polygonaceae. Pungent flavor and warm nature. It enters lung meridian, spleen meridian and liver meridian. Has effects in warming middle energizer, promoting diuresis, removing blood stasis, and resolving hard mass, and can be used for treating vomiting, diarrhea, abdominal pain, abdominal mass, distention, edema, carbuncle, swelling, skin and external diseases, and lymphoid tuberculosis. The record of Shen nong Ben Cao Jing: "improving eyesight, warming middle energizer, resisting wind-cold, expelling pathogenic wind-cold, edema of face and eyes, carbuncle and ulcer. In the 'materia medica harm good' records polygonum multiflorum as guiding drugs of fructus corni and fructus evodiae in liver tonifying and liver warming. Modern physicians Zhang Shouyi said: polygonum persicum, which has strong force and is most liable to abortion, is essential for pregnant women; there are also deficiency of blood and qi, and the patient is unsmooth in the month, which is not caused by stasis. The journal of practical traditional Chinese medicine department records that the polygonum frutescens can be added to achieve better treatment effect in the middle-stage cirrhosis after hepatitis treatment. The Polygonum hydropiper fruit is used as a ministerial drug, not only can the effects of treating liver be better played by the Rangoon creeper fruit, but also has the effects of removing stasis and eliminating stagnation, and the Polygonum hydropiper fruit and the Rangoon creeper fruit mutually act to calm the liver and fix the liver; it can also be used for treating vomiting and diarrhea with high abdominal pain, and can relieve abdominal viscera.
Adjuvant drugs: thlaspi arvense seed, pungent and bitter in flavor, slightly warm in nature. It enters liver, kidney and spleen meridians. For treating lung heat, liver heat, kidney vessel damage, testicle swelling and dropping, seminal emission, yang fistula, lumbago, skelalgia, nausea, etc., the records in Benjing are: the pennycress with pungent flavor, slightly warm and nontoxic. Has effects in improving eyesight, relieving eye pain, removing obstruction of qi in the eyes, tonifying five internal organs, and improving essence and light. Treat low back pain of heart and abdomen. It is suitable for long-term use for reducing weight and resisting aging. The 'Thlaspis arvense' recorded in the 'medical treatise on liver accumulation and red and swollen eyes' in the 'medical property treatise'. Record in the famous physicians bibliography: ' Thlaspi arvense seed is nontoxic and treats heart and abdominal lumbago. Thlaspis arvense is recorded in food materia medica, and can clear liver, improve eyesight and treat white diarrhea caused by intestinal deficiency and cold air invasion. Modern researches find that the pennycress mainly contains volatile oil, reducing sugar and various enzymes, has the effects of resisting depression, tumors and nerve oxidative damage, can assist monarch drug in relieving liver heat, preventing and treating liver diseases, clearing heat and removing toxicity, clearing liver and improving eyesight, and cooperates with monarch drug fiveleaf akebia to dredge stagnated liver qi. Meanwhile, the traditional Chinese medicine composition has the effects of conditioning intestinal tracts, stimulating the appetite and tonifying the spleen, complementing the defects of other internal organs, playing a role of assisting monarch and minister medicines and serving as an adjuvant medicine.
A messenger drug: gou Gu Zi, named Gong Lao Zi. Fructus Ilicis Cornutae Immaturum has effects of nourishing yin, replenishing vital essence, promoting blood circulation, and removing blood stasis, and can be used as nourishing and detoxicating medicine for treating yin deficiency and internal heat. The south medicine "Chinese herbal medicine" mentions that: bitter, astringent and slightly warm. It enters liver, kidney and spleen meridians, tonifies liver and kidney, strengthens tendons and activates collaterals, and astringes and controls lower energizer. It can be used for treating asthenia, low fever, arthralgia and myalgia, metrorrhagia, leukorrhagia, and diarrhea. The original menstrual flow is recorded in the original book: it is a special medicine for promoting blood circulation, removing blood stasis and strengthening muscles and bones. It is also the essential herb for replenishing marrow and astringing essence and blood. "Jiangsu province plant medicine records: it is indicated for yin deficiency with internal heat, and is used as a nourishing and heat-relieving herb, and has the same action as Zhi Zhen Zi. The Chinese herbal medicines commonly used in Shanghai are mentioned as follows: to tonify liver and kidney, stop diarrhea. It is indicated for asthenia with low fever, metrorrhagia, and diarrhea. Modern researches show that the Chinese holly fruit mainly contains chemical components such as fatty oil, alkaloid, saponin, tannin, bitter substance and the like, and has the effects of resisting carcinogenesis, promoting cancer, resisting angiogenesis, promoting blood circulation, removing blood stasis, eliminating dampness and relieving pain. The Chinese holly fruit, as a guiding drug, has the effect of harmonizing the drugs and eliminating pathogens. The other medicines have the main effects of promoting blood circulation to remove blood stasis, clearing damp and strengthening spleen, and breaking blood stasis and dissipating stagnation, and the Chinese holly fruit is used as a relative yin nourishing medicine, so that yin can be prevented from being excessively injured by other medicines and damaging yin fluid while the blood circulation is promoted and the blood stasis is dissipated.
In conclusion, fiveleaf akebia fruit, as a monarch drug, has the main effects of soothing the liver and harmonizing the stomach, promoting blood circulation to arrest pain, and softening and resolving hard mass in the compound preparation. Tang Dynasty famous physicians Meng Shen: "Thick intestines and stomach" is good for people to eat, lower triple energizer, remove bad smell and make children eat better. Unblock the twelve main meridians. It can be seen that fiveleaf akebia fruit not only plays a main role in treating hepatic fibrosis, but also has the effects of regulating qi, harmonizing stomach, eliminating dampness and tonifying spleen, and has a certain conditioning effect on the whole human body, and is used as a monarch drug. The polygonum multiflorum thumb is used as a ministerial drug, is nontoxic, mainly plays a role in removing stasis and dissipating stagnation in the compound preparation, has the functions of dissipating heat and removing food retention, warming the middle warmer, resisting wind cold and the like, can effectively solve the problem of poor appetite caused by eating the drug, warms and nourishes the spleen and stomach and improves the immunity of a human body, and is used as the ministerial drug. The pennycress is mild in medicine property and has the effects of tonifying kidney, benefiting qi and harmonizing stomach, the problem that the kidney is damaged due to long-term taking of the traditional Chinese medicine can be solved, and the whole medicine becomes mild in medicine property and reduces side effects on a human body while assisting monarch medicine to dredge liver stasis and strengthen spleen and stomach. The Chinese holly fruit is used as a guiding drug and can play a role in harmonizing various drugs, the Chinese holly fruit is matched with the monarch drug fiveleaf akebia fruit to treat main symptoms of the Chinese holly fruit except for treating the accompanying symptoms in the prescription, the curative effect of the monarch drug fiveleaf akebia fruit is improved, and in addition, the Chinese holly fruit is used as a relative yin nourishing drug and can prevent yin injury while promoting blood circulation and removing blood stasis.
In a word, the traditional Chinese medicine compound preparation is prepared on the basis of 'the human body is an organic whole', reasonably applies the theories of 'treatment based on syndrome differentiation', 'holism concept' and the like and according to the principle of monarch, minister, assistant and guide drug administration, and has clear components and accurate efficacy. The preparation is non-toxic and harmless to a human body, has a remarkable treatment effect on hepatic fibrosis, allows a hepatic fibrosis patient to have a larger space for selecting a medicament, gives more treatment hopes to the patient, brings certain economic benefits to related enterprises, and promotes the rapid development of related industries. Meanwhile, the research and development of the preparation also promotes the traditional Chinese medicine culture to a certain extent, so that more people can accept and use the medical product.
Drawings
FIG. 1 shows the effect of a Chinese herbal compound preparation on the serum liver injury index of a fibrotic rat, wherein FIG. 1-1 shows the effect of the Chinese herbal compound preparation on serum ALT, FIG. 1-2 shows the effect of the Chinese herbal compound preparation on serum AST, FIG. 1-3 shows the effect of the Chinese herbal compound preparation on serum ALP, and FIG. 1-4 shows the effect of the Chinese herbal compound preparation on serum hydroxyproline;
FIG. 2 shows the effect of the compound Chinese medicinal preparation on the liver fibrosis index of the fibrosis rat serum, wherein FIG. 2-1 shows the effect of the compound Chinese medicinal preparation on the serum LN, FIG. 2-2 shows the effect of the compound Chinese medicinal preparation on the serum HA, FIG. 2-3 shows the effect of the compound Chinese medicinal preparation on the serum IV-C, and FIG. 2-4 shows the effect of the compound Chinese medicinal preparation on the serum PC-III;
FIG. 3 shows the effect of the compound Chinese medicine preparation on the liver structure of a rat with liver fibrosis;
FIG. 4 shows the effect of the compound Chinese medicine preparation on collagen deposition in liver tissue of liver fibrosis rat.
Detailed Description
The preparation of the compound Chinese medicinal preparation and the efficacy of treating hepatic fibrosis are further illustrated by the following examples.
Example 1: preparation of oral liquid
Taking 30g of fiveleaf akebia fruit, 20g of polygonum multiflorum, 10g of pennycress and 5g of Chinese holly fruit, adding 10 times of 60% ethanol, heating, refluxing and extracting for 3 times, each time for 2 hours, filtering, combining the filtrates, concentrating under reduced pressure (the temperature is 60-70 ℃, the pressure is 0.08 Mpa) to the relative density of 1.05-1.10 (the temperature is 60-70 ℃), adding 1% aspartame and 2% citric acid, stirring uniformly, standing for more than 12 hours, filtering, subpackaging and sterilizing to obtain the traditional Chinese medicine.
Example 2: capsule preparation
20g of fiveleaf akebia fruit, 30g of polygonum fruit, 5g of pennycress and 8g of Chinese holly fruit are taken, 10 times of 60% ethanol is added, and the mixture is heated and refluxed for extraction for 3 times, and each time lasts for 2 hours. Mixing the extractive solutions, recovering ethanol, concentrating, evaporating to dryness, drying under reduced pressure (60-70 deg.C and 0.08 mpa), pulverizing into fine powder, adding ethanol, granulating, drying, and making into capsule.
Example 3: preparation of granules
40g of fiveleaf akebia fruit, 10g of polygonum multiflorum, 15g of pennycress and 3g of Chinese holly fruit are taken, 10 times of 60% ethanol is added, heating reflux extraction is carried out for 3 times, 2 hours of each time, filtration is carried out, filtrate is combined, reduced pressure concentration (the temperature is 60-70 ℃ and the pressure is 0.08 Mpa) is carried out until the relative density is 1.05-1.10 (the temperature is 60-70 ℃), 2% aspartame is added, stirring is carried out, spray drying is carried out, dry granulation is carried out, and subpackaging is carried out by an aluminum-plastic composite membrane to obtain the traditional Chinese medicine.
Example 4: research on pharmacological action of Chinese herbal compound preparation for treating hepatic fibrosis
Animal model preparation and dosing regimen: 50 male SD rats (with the weight of 200g-220 g) are randomly divided into a normal control group, a model group and low, medium and high dose groups (150 mg/kg, 300mg/kg and 600mg/kg oral liquid) of the traditional Chinese medicine compound preparation prepared in the embodiment 1 of the invention, and each group comprises 10 rats. After the rats are sent to a GLP experimental animal center, free food and water are supplied to the rats, the rat feed meets the Chinese standard (LAD 1000), and the rat water meets the requirements of environmental and facility of experimental animals (GB 14925-2010). Rats are given 14 hours of light and 10 hours of standard light cycle in the dark, the room temperature is controlled between 18-26 ℃ relativelyHumidity is maintained at 40-70%, noise is controlled below 85 dB, ammonia concentration is maintained below 20PPm, and ventilation is carried out for 8-12 times/hr. All rats were acclimatized for 1 week before subsequent experiments. Mixing CCl 4 Mixing with olive oil at a volume of 1:1 to 50% 4 The solution is used for establishing a hepatic fibrosis model. 150mg/kg, 300mg/kg, 600mg/kg of a herbal compound oral liquid was prepared for drug treatment as described in example 1. The remaining 4 groups of rats except the normal control group were intraperitoneally injected at a dose of 1ml/kg 50% CCl 4 Solution, 3 times per week for 8 weeks, normal control rats were injected intraperitoneally with the same volume of olive oil at the same time point, at a similar location, as a blank. Intraperitoneal injection of 50% CCl 4 After 4 weeks of the solution, the low, medium and high doses of the compound traditional Chinese medicine preparation are respectively infused into the stomach according to the doses of 150mg/kg, 300mg/kg and 600mg/kg, and the traditional Chinese medicine compound preparation is infused 1 time per day for 4 weeks. After all experiments were completed, each rat was weighed and recorded for anesthesia by intraperitoneal injection of 50mg/kg pentobarbital solution following strict standard protocol (SOP) for experimental animal sacrifice (rats, rabbits), blood was rapidly collected using a 0.05mm inner diameter capillary, then the liver was separated by dissection of the abdominal cavity, a picture of the complete liver of each rat was taken using a camera, and each liver was weighed and recorded using an analytical balance. After the collected blood sample is kept still for 2 hours at room temperature, the blood sample is centrifuged for 15 minutes at 3500 rpm by using a centrifuge, and the supernatant is sucked and stored at the temperature of minus 20 ℃ for subsequent serological index detection. A small portion of each rat liver was fixed in 10% formalin for subsequent histopathological examination. We find that the traditional Chinese medicine compound preparation can obviously reduce the levels of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase of blood serum of a rat in a hepatic fibrosis model, reduce the collagen deposition of a fibrotic liver and improve the pathological injury of the fibrotic liver, and the result is shown in the attached figures 1-4 in detail.
4.1 Effect of Compound preparation of Chinese herbs on fibrosis rat serum liver injury index
The biochemical instrument is used for detecting the fibrosis rat serum liver injury index. The results show that the serum ALT level of the normal control group rat is 149 +/-1.8U/L, the AST level is 34 +/-2.1U/L, the ALP level is 217 +/-2.4U/L, the hydroxyproline level is 21 +/-3.2U/L, the liver injury indexes are all at lower levels, the serum ALT level of the model group rat is 431 +/-2.4U/L, the AST level is 191 +/-2.6U/L, the ALP level is 514 +/-2.9U/L, the hydroxyproline level is 78 +/-1.7U/L, the liver injury indexes are all obviously increased, the traditional Chinese medicine compound preparation can dose-dependently reduce the serum ALT, AST, ALP and hydroxyproline levels of the fibrosis rat, the serum ALT level of the high-dose group rat is 173 +/-2.5U/L, the AST level is 47 +/-2.3U/L, the ALP level is 257 +/-1.8U/L, the hydroxyproline level is 22 +/-3.1U/L, and the traditional Chinese medicine compound preparation can improve the liver injury indexes (effective graph).
Table 1: the detection results (x + -s) of the serum liver injury indexes ALT, AST, ALP and HYP of the rats in each group
(in comparison with the normal control group, ### P<0.001, P compared to model group<0.05,**P<0.01,***P<0.001)
4.2 Effect of Compound preparation of Chinese herbs on fibrosis rat serum hepatic fibrosis index
The kit is used for detecting the hepatic fibrosis marker of the serum of the fibrosis rat. The results show that the serum LN level of normal control group rats is 51 + -2.4 mug/L, the HA level is 109 + -4.1 mug/L, the IV-C level is 18 + -2.7 mug/L, the PC-III level is 23 + -1.8 mug/L, the liver fibrosis index is all at a lower level, the serum LN level of model group rats is 169 + -4.7 mug/L, the HA level is 328 + -5.5 mug/L, the IV-C level is 61 + -3.6 mug/L, the PC-III level is 73 + -2.1 mug/L, the liver fibrosis index is all significantly increased, the Chinese herbal compound preparation can dose-dependently reduce the fibrosis rat serum LN, HA, IV-C, PC-III level, the high dose group rats serum level is 56 + -3.4 mug/L, the HA level is 126 + -1.8U/L, the IV-C level is 22 + -1.3U/L, the PC-III level is 25U/L, and the pathological damage level is effectively improved by the Chinese herbal compound preparation (the pathological damage is shown in the picture).
Table 2: the liver fibrosis indexes LN, HA, IV-C, PC-III of each group of rats (x + -s)
(in comparison with the normal control group, ### P<0.001, P compared to model group<0.05,**P<0.01,***P<0.001)
4.3 Effect of Compound preparation of Chinese traditional medicine on liver Structure of hepatic fibrosis rat
Taking the livers of rats in each group in the animal experiment, and observing by macroscopic observation, the livers of the rats in a normal group are bright red and have no tissue lesion, while the livers of the rats in a model group are whitish and have severe fibrosis tissue lesion; the Chinese medicinal compound preparation can improve CCl dose-dependently 4 Causing pathological changes of liver tissues of rats, wherein the improvement degree of the high-dose traditional Chinese medicine compound preparation prepared in the embodiment 1 of the invention on the liver tissue injuries is most obvious. We also performed histopathological examination of groups of rat livers using HE staining. The results show that the liver lobules of the rats in the normal group have complete structures, the liver cells are arranged regularly, and no inflammatory cell infiltration exists; the liver of the rat in the model group has pseudolobule, the arrangement of liver cells is disordered, the fibrous connective tissue hyperplasia is obvious, and inflammatory cell infiltration is serious; in the treatment group of the traditional Chinese medicine compound preparation, with the increase of the administration dosage, the false lobules of the rat liver gradually disappear, the liver cells gradually recover and are arranged regularly, the fibrous connective tissue is obviously reduced, and the inflammatory cell infiltration basically disappears, which indicates that the traditional Chinese medicine compound preparation can obviously improve the structural change of the fibrotic liver (figure 3).
4.4 the effect of the compound Chinese medicine preparation on the collagen deposition in the liver tissue of the liver fibrosis rat.
Taking rat livers of each group in the animal experiment, and examining the improvement effect of the traditional Chinese medicine compound preparation on the collagen deposition of the fibrosis liver by using Sirius red staining and massson staining. The liver tissue fixed in 10% formalin solution was taken out, placed in a fume hood to select an appropriate target position, carefully separated into appropriate size tissue blocks with a scalpel, placed in a dehydrator for dehydration with gradient alcohol in sequence, the dehydrated tissue blocks were placed in an embedding machine for standardized paraffin embedding, and then the wax blocks were placed on a paraffin microtome for sectioning with a slice thickness of 4 μm. The tissue was fixed with a glass slide and placed in a 60 ℃ oven for baking. Taking out after water baking and wax baking and roasting for standby at normal temperature. Tissue sections were subjected to conventional Sirius red staining and massson staining to investigate liver collagen deposition. The results show that the liver of the normal group of rats has no collagen deposition basically, the liver of the model group of rats has severe collagen deposition, and the Chinese herbal compound preparation with different doses can reduce the collagen deposition, which indicates that the Chinese herbal compound preparation can reduce the collagen deposition of the fibrosis liver (figure 4).
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. A traditional Chinese medicine compound preparation for treating hepatic fibrosis is characterized by being prepared from the following raw material medicines in parts by weight: 20-40 parts of fiveleaf akebia fruit, 10-30 parts of Polygonum flaccidum fruit, 5-15 parts of Thlaspis arvense seed and 3-8 parts of Chinese holly fruit.
2. The compound traditional Chinese medicine preparation for treating hepatic fibrosis as claimed in claim 1, which is prepared from the following raw material medicines in parts by weight: 20 parts of fiveleaf akebia fruit, 30 parts of Polygonum multiflorum fruit, 5 parts of Thlaspi arvense and 8 parts of Chinese holly fruit.
3. The compound traditional Chinese medicine preparation for treating hepatic fibrosis as claimed in claim 1, which is prepared from the following raw material medicines in parts by weight: 40 parts of fiveleaf akebia fruit, 10 parts of Polygonum flaccidum fruit, 15 parts of Thlaspi arvense seed and 3 parts of Chinese holly fruit.
4. The compound traditional Chinese medicine preparation for treating hepatic fibrosis as claimed in claim 1, which is prepared from the following raw material medicines in parts by weight: 30 parts of fiveleaf akebia fruit, 20 parts of Polygonum flaccidum fruit, 10 parts of Thlaspis arvense seed and 5 parts of Chinese holly fruit.
5. The compound Chinese medicinal preparation for treating hepatic fibrosis according to claim 1, wherein the preparation method comprises the following steps: collecting fructus Akebiae, fructus Polygoni Caespitosi, semen Thlaspis, and fructus Ilicis Cornutae, adding 10 times of 60% ethanol, heating and reflux-extracting for 3 times, each for 2 hr, mixing extractive solutions, recovering ethanol, adding appropriate adjuvant, and making into oral liquid by conventional method.
6. The compound Chinese medicinal preparation for treating hepatic fibrosis according to claim 1, wherein the preparation method comprises the following steps: taking fiveleaf akebia fruit, polygonum fruit, pennycress and Chinese holly fruit, adding 10 times of 60% ethanol, heating and refluxing for 3 times, extracting for 2 hours each time, combining extracting solutions, recovering ethanol, concentrating and evaporating to dryness, drying under reduced pressure, crushing into fine powder, adding appropriate auxiliary materials, and preparing into granules, tablets, capsules or micro pills by using a conventional preparation method.
7. The compound Chinese medicinal preparation for treating hepatic fibrosis according to claim 6, wherein the drying under reduced pressure in the preparation method is performed at 60-70 deg.C and under 0.08mpa.
8. The use of the compound Chinese medicinal preparation of claim 1 in the preparation of a medicament for treating hepatic fibrosis.
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| CN103083573A (en) * | 2013-01-20 | 2013-05-08 | 崔英坤 | Traditional Chinese medicine used for treating alcoholic fatty liver |
| CN103393857A (en) * | 2013-08-19 | 2013-11-20 | 张成娟 | Drug for curing hepatitis and preparation method thereof |
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| CN102961564A (en) * | 2012-12-16 | 2013-03-13 | 陈淑香 | Pharmaceutical composition for treating alcoholic cirrhosis |
| CN103083573A (en) * | 2013-01-20 | 2013-05-08 | 崔英坤 | Traditional Chinese medicine used for treating alcoholic fatty liver |
| CN103393857A (en) * | 2013-08-19 | 2013-11-20 | 张成娟 | Drug for curing hepatitis and preparation method thereof |
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