Pharmaceutical composition and pharmaceutical preparation for improving non-alcoholic steatohepatitis and application

Abstract

The invention relates to the technical field of traditional Chinese medicines, and discloses a pharmaceutical composition for improving non-alcoholic steatohepatitis, which is prepared from the following components in parts by weight: 10 to 60 parts of radix bupleuri, 0 to 30 parts of radix scutellariae, 0 to 90 parts of radix salviae miltiorrhizae, 0 to 30 parts of cassia twig, 0 to 30 parts of rhizoma zingiberis, 0 to 45 parts of fructus crataegi, 0 to 45 parts of radix trichosanthis, 0 to 90 parts of raw oyster shell and 0 to 20 parts of honey-fried licorice root. The pharmaceutical composition can improve liver fatty degeneration, inflammation and fibrosis, reduce accumulation of liver TG, reverse increase of ALT, AST and ALP in serum, reduce the liver weight and liver index of non-alcoholic steatohepatitis and play a role in liver protection. The invention can provide drug choices for the preparation of drugs for treating non-alcoholic steatohepatitis.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and in particular to a pharmaceutical composition and a pharmaceutical preparation for improving non-alcoholic fatty hepatitis and applications thereof. Background Art

[0002] Non-alcoholic fatty liver disease (NAFLD) refers to a clinical pathological syndrome characterized by fat accumulation in liver cells other than alcohol and other clear interfering factors. Non-alcoholic steatohepatitis (NASH) is a late form of NAFLD, characterized by fatty degeneration, inflammation and fibrosis of the liver. Modern epidemiological evidence shows that the incidence of NASH is increasing year by year and is about to surpass viral hepatitis to become the most important cause of cirrhosis and liver cancer. In the early stages of NASH before the development of liver fibrosis, lifestyle interventions including weight loss, exercise, and dietary changes are the main treatment strategies. When liver fibrosis occurs, blood sugar-lowering, lipid-lowering and liver-protecting drugs are often used clinically to control the progression of NASH. However, these drugs have no definite effect on NASH and liver fibrosis, and have large adverse reactions. Currently, only Saroglitazar and Resmetirom (trade name Rezdiffra) can be used to treat NASH in the world. Saroglitazar was approved by the Indian Drug Administration for the treatment of NASH in March 2020, but this indication has not yet been approved by the US FDA; Resmetirom was approved by the FDA for the treatment of NASH in March 2024, but it is expensive and can cause liver damage and gallbladder-related adverse reactions. NASH is a chronic metabolic disease with a complex pathogenesis involving lipid metabolism disorders, inflammatory responses, and intestinal flora imbalances. It is difficult for a single target drug to fully solve the problem, and a large number of NASH patients are still facing a situation where there is no medicine to treat them.

[0003] TCM treatment of diseases follows the principle of syndrome differentiation and treatment. Non-alcoholic fatty hepatitis belongs to the category of "liver disease", "flank pain" and "accumulation" in TCM. TCM believes that improper diet, improper work and rest, emotional disorder, long-term illness and physical weakness, and insufficient endowment are the main causes of the disease. The disease is located in the liver, involving the spleen and kidney. Its pathological basis is related to phlegm, dampness, turbidity, blood stasis, heat, etc. The syndrome belongs to the deficiency of the root and the excess of the symptoms, with spleen deficiency as the root and phlegm turbidity and blood stasis as the symptoms. NASH can be divided into four TCM syndromes: liver depression and spleen deficiency syndrome, the treatment method is mainly to soothe the liver and strengthen the spleen; phlegm and dampness internal obstruction syndrome, the treatment method is to strengthen the spleen and replenish qi, resolve phlegm and remove dampness; damp-heat accumulation syndrome, the treatment method is to clear heat and remove dampness; phlegm and blood stasis syndrome, the treatment method is to activate blood circulation and remove blood stasis, and resolve phlegm and disperse the knot. The "Expert Consensus on Diagnosis and Treatment of Non-alcoholic Fatty Liver Disease in Traditional Chinese Medicine (2023)" recommends Xiaoyao Pills, Qianggan Capsules, Shugankang Capsules, etc. for the treatment of non-alcoholic fatty liver disease. These drugs are not specific medicines for the treatment of non-alcoholic fatty liver disease, and there is currently no specific Chinese medicine for non-alcoholic fatty liver disease.

[0004] The traditional Chinese medicines currently used in clinical treatment of non-alcoholic fatty liver disease include Xiaoyao Pills, Qianggan Capsules, Shugankang Capsules, etc. Among them, Xiaoyao Pills (Bupleurum, Angelica, White Peony, Stir-fried Atractylodes, Poria, Roasted Licorice, and Menthol) soothe the liver and strengthen the spleen, nourish blood and regulate menstruation, and are mainly used for depression, chest and flank pain caused by liver depression and spleen deficiency; Qianggan Capsules (Artemisia capillaris, Isatis root, Angelica, White Peony, Salvia miltiorrhiza, Curcuma, Astragalus, Codonopsis, Alisma, Polygonatum, Rehmannia root, Chinese yam, hawthorn, Liushenqu, qinjiao, licorice) clear away heat and dampness, nourish the spleen and blood, invigorate qi and relieve depression, and are used for chronic hepatitis, early liver cirrhosis, fatty liver, toxic hepatitis, etc. Shugankang capsule (astragalus, codonopsis, ophiopogon, angelica, white peony root, bupleurum, scutellaria, curcuma, fructus aurantii, poria, licorice, schisandra) invigorates qi and nourishes yin, softens the liver and strengthens the spleen, and is used for irritability, fatigue, loss of appetite, and chest and flank pain caused by liver depression and spleen deficiency. The above Chinese medicines can be used as adjuvant treatment for NASH, but their main indications are not NASH, which is not completely consistent with the pathogenesis of NASH, "phlegm, turbidity, and blood stasis". Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the pharmaceutical composition and pharmaceutical preparation for improving non-alcoholic fatty hepatitis and their application provided by the present invention provide a medication option for the treatment of non-alcoholic fatty hepatitis.

[0006] In order to achieve the above-mentioned purpose of the invention, the technical solution adopted by the present invention is: a pharmaceutical composition for improving non-alcoholic fatty hepatitis, the pharmaceutical composition is composed of the following components in weight ratio: 10-60 parts of bupleurum, 0-30 parts of scutellaria, 0-90 parts of salvia miltiorrhiza, 0-30 parts of cinnamon twig, 0-30 parts of dried ginger, 0-45 parts of hawthorn, 0-45 parts of trichosanthes, 0-90 parts of raw oyster, and 0-20 parts of roasted licorice.

[0007] Furthermore, the above-mentioned pharmaceutical composition for improving non-alcoholic fatty hepatitis is composed of the following components in the following weight ratio: 20-60 parts of bupleurum, 10-30 parts of scutellaria, 30-90 parts of salvia miltiorrhiza, 10-30 parts of cinnamon twig, 10-30 parts of dried ginger, 15-45 parts of hawthorn, 15-45 parts of trichosanthes, 30-90 parts of raw oysters, and 10-20 parts of roasted liquorice.

[0008] Furthermore, a pharmaceutical preparation for improving non-alcoholic fatty hepatitis, the active ingredient of the pharmaceutical preparation is the above-mentioned pharmaceutical composition.

[0009] Furthermore, the pharmaceutical preparation for improving non-alcoholic fatty hepatitis is a pharmaceutical preparation prepared by using an extract after mixing and extracting the pharmaceutical composition in proportion.

[0010] Furthermore, the above-mentioned pharmaceutical preparation is used to improve diseases or symptoms related to non-alcoholic fatty liver disease.

[0011] Furthermore, the above-mentioned pharmaceutical preparation is used to improve diseases or symptoms related to non-alcoholic fatty liver disease, and the symptoms of non-alcoholic fatty liver disease include liver fatty degeneration, liver inflammation and liver fibrosis, liver function damage and liver lipid accumulation.

[0012] The beneficial effects of the present invention are as follows: the saikosaponin compounds contained in bupleurum have anti-inflammatory and immunomodulatory effects, and can reduce the inflammatory response of the liver. Bupleurum can also reduce the accumulation of fat in liver cells by regulating the lipid metabolism pathway of the liver, thereby improving the symptoms of fatty liver.

[0013] Flavonoids such as baicalin and wogonin in Scutellaria baicalensis can reduce inflammation and oxidative stress in the liver.

[0014] Danshen has the effects of promoting blood circulation and removing blood stasis, and improving microcirculation. Its main active ingredients, tanshinone and tanshinone, can protect the liver by promoting liver cell regeneration and inhibiting liver fibrosis. Danshen can downregulate the TGF-β signaling pathway, thereby reducing the degree of liver tissue fibrosis.

[0015] Cinnamon twig has the functions of warming the meridians and promoting yang and transforming qi. Its volatile oil components can promote blood circulation, enhance the body's immune function, and may help reduce liver fat accumulation by improving insulin sensitivity.

[0016] Gingerol, the active ingredient in dried ginger, has antioxidant and anti-inflammatory effects, which can reduce oxidative stress and inflammation in the liver and further protect liver cells.

[0017] Hawthorn is rich in flavonoids and triterpenoids, which have lipid-lowering, anti-oxidation and lipid-improving effects. Hawthorn extract can reduce liver lipid deposition by regulating the expression of genes related to lipid metabolism.

[0018] Radix Trichosanthis has the effects of clearing away heat, promoting the production of body fluid, reducing swelling and dispersing nodules. Its active ingredient, trichosanthesin, may alleviate liver inflammation through anti-inflammatory and immunomodulatory effects.

[0019] Raw oysters are rich in trace elements such as zinc and selenium, have antioxidant and immune-enhancing effects, help protect liver cells from free radical damage, and inhibit the occurrence of liver fibrosis to a certain extent.

[0020] The main components of licorice, glycyrrhizic acid and glycyrrhetinic acid, have significant anti-inflammatory and immunomodulatory effects, and can inhibit liver inflammation and fibrosis through multiple signaling pathways.

[0021] This drug combination can improve liver function damage by reducing liver weight and liver index of non-alcoholic fatty liver disease, reversing the increase of ALT, AST and ALP in serum, reducing liver lipid accumulation, improving liver fatty degeneration, downregulating the expression of inflammatory factors, and regulating the expression of profibrotic factors in the liver, thereby improving non-alcoholic fatty liver disease and related diseases or symptoms.

[0022] The present invention uses bupleurum, scutellaria and salvia miltiorrhiza as the main drugs. Bupleurum soothes the liver and relieves depression, scutellaria clears away heat and detoxifies, and salvia miltiorrhiza promotes blood circulation and removes blood stasis. The three are combined to achieve the functions of soothing the liver, promoting blood circulation and reducing turbidity. The assistant drugs cinnamon twig, dried ginger and hawthorn warm the middle jiao, promote blood circulation, remove blood stasis and reduce fat. The adjuvant drugs trichosanthes and raw oyster can relieve the deficiency of body fluid caused by qi stagnation and heat in the liver and gallbladder meridians, water retention and phlegm and dampness obstruction. Compared with the traditional Chinese medicine currently used in clinical treatment of non-alcoholic fatty liver disease, this prescription has different drug composition and treatment method, is more in line with the pathogenesis of "phlegm, turbidity and blood stasis" of NASH, and has great potential to be developed into a drug for the treatment of NASH. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The figures are the effects of the pharmaceutical composition on the liver pathological tissues of NASH rats after HE (Hematoxylin-Eosin) staining, Oil Red O (Oil Red O) staining and Masson (Masson) staining respectively;

[0024] Figure 2 This is a diagram showing the effect of the pharmaceutical composition on liver pathological tissue (HE staining) of NASH rats;

[0025] Figure 3 This is a graph showing the effect of the pharmaceutical composition on liver fatty degeneration, lobular inflammation and ballooning degeneration scores and NAS comprehensive score in liver pathological tissue (HE staining) of NASH rats;

[0026] Figure 4 This is a diagram showing the effect of the pharmaceutical composition on liver pathological tissue (Oil Red O staining) of NASH rats;

[0027] Figure 5 This is an analysis diagram of the Oil Red O staining area of ​​the liver pathological tissue of NASH rats;

[0028] Figure 6 This is a diagram showing the effect of the pharmaceutical composition on liver pathological tissue (Masson staining) of NASH rats;

[0029] Figure 7 This is a diagram of fibrosis area analysis of liver pathological tissue of NASH rats under the influence of Masson staining;

[0030] Figure 8This is a graph showing the effect of the pharmaceutical composition on the mRNA expression levels of liver inflammatory factors TNF-α, IL-6, IL-1β, and MCP-1 in NASH rats;

[0031] Fig. 9 This graph shows the effect of the pharmaceutical composition on the mRNA expression levels of liver fibrosis factors TGFβ, Acta2, Col1a1, and Col1a2 in NASH rats. DETAILED DESCRIPTION

[0032] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0033] The present embodiment provides a pharmaceutical composition for improving non-alcoholic fatty hepatitis, the pharmaceutical composition is composed of the following components in weight ratio: 10-60 parts of bupleurum, 0-30 parts of scutellaria, 0-90 parts of salvia miltiorrhiza, 0-30 parts of cinnamon twig, 0-30 parts of dried ginger, 0-45 parts of hawthorn, 0-45 parts of trichosanthes, 0-90 parts of raw oyster, 0-20 parts of licorice. Preferably, the pharmaceutical composition is composed of the following components in weight ratio: 20-60 parts of bupleurum, 10-30 parts of scutellaria, 30-90 parts of salvia miltiorrhiza, 10-30 parts of cinnamon twig, 10-30 parts of dried ginger, 15-45 parts of hawthorn, 15-45 parts of trichosanthes, 30-90 parts of raw oyster, 10-20 parts of licorice.

[0034] The pharmaceutical composition of the present invention can improve liver damage caused by non-alcoholic fatty hepatitis, lower liver index, reduce the accumulation of triglycerides (TG) in the liver, alleviate liver fatty degeneration and inflammation, and inhibit liver fibrosis. Therefore, the present invention can lay a working foundation for the clinical drug development of non-alcoholic fatty hepatitis.

[0035] According to a typical embodiment of the present invention, the pharmaceutical preparation is a finished product prepared by mixing raw materials in proportion and then extracting the extract; optionally, the pharmaceutical preparation is a finished drug prepared by mixing the extract with excipients in proportion and then processing the extract into the extract.

[0036] In a typical embodiment of the present invention, the pharmaceutical preparation further includes pharmaceutically acceptable excipients, and the dosage form of the pharmaceutical preparation can be tablets, granules, capsules, pills, suppositories, powders, ointments, drops, aerosols, powder sprays, solutions, suspensions, syrups, mixtures, wine preparations, tea preparations, lozenges, freeze-dried powder injections, or emulsions; it can be a conventional preparation, a sustained-release preparation, a controlled-release preparation, and various microparticle delivery systems.

[0037] The pharmaceutical preparation of the present invention containing an effective dose can be prepared using pharmaceutical carriers familiar to those skilled in the art. For example, oral preparations (such as tablets, capsules, solutions or suspensions). Pharmaceutical preparation carriers include: binders used in oral preparations (such as starch, usually corn, wheat or rice starch, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinyl pyrrolidone), diluents (such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycerol), lubricants (such as silicon dioxide, talc, stearic acid or its salt, usually magnesium stearate or calcium stearate, and / or polyethylene glycol), and if necessary, disintegrants such as starch, agar, alginic acid or its salt, usually sodium alginate, and / or effervescent mixtures, solubilizers, stabilizers, suspending agents, pigments, flavoring agents, etc., preservatives, solubilizers, stabilizers, etc. used in injectable preparations; matrix, diluent, lubricant, preservative, etc. used in topical preparations. Pharmaceutical preparations can be administered orally and if certain drugs are unstable under gastric conditions they can be formulated as enteric coated tablets.

[0038] The term "effective dose" of the pharmaceutical preparation of the present invention refers to a sufficient amount of the compound to treat the disorder at a reasonable effect / risk ratio applicable to any medical treatment and / or prevention. However, it should be recognized that the total daily dose of the pharmaceutical preparation of the present invention must be determined by the attending physician within the scope of sound medical judgment. For any specific patient, the specific therapeutically effective dose level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex and diet; the administration time, route of administration and excretion rate of the specific compound used; the duration of treatment; drugs used in combination or concomitantly with the specific compound used; and similar factors known in the medical field. For example, the practice in the art is to start the dosage of the Chinese medicine pharmaceutical composition from a level lower than that required to obtain the desired therapeutic effect and gradually increase the dosage until the desired effect is obtained.

[0039] Example 1

[0040] Preparation of pharmaceutical composition Y1:

[0041] Weigh by weight 18 parts of bupleurum, 9 parts of scutellaria, 30 parts of salvia miltiorrhiza, 10 parts of cinnamon twig, 10 parts of dried ginger, 15 parts of hawthorn, 15 parts of trichosanthes, 30 parts of raw oyster, and 6 parts of roasted liquorice.

[0042] The specific process for preparing the pharmaceutical composition is as follows: weigh a corresponding amount of Chinese herbal medicine slices, add 10 times the amount of water and boil twice, each time for 1 hour, combine the decoctions, filter, concentrate the filtrate into a thick paste, dry under reduced pressure into a dry paste powder, crush, and prepare a solution of corresponding concentration before use.

[0043] Example 2

[0044] Preparation of pharmaceutical composition Y2:

[0045] Weigh by weight 18 parts of bupleurum, 9 parts of scutellaria, 10 parts of cinnamon twig, 10 parts of dried ginger, 15 parts of trichosanthes, 30 parts of raw oyster, and 6 parts of roasted liquorice.

[0046] The specific process for preparing the pharmaceutical composition is as follows: weigh a corresponding amount of Chinese herbal medicine slices, add 10 times the amount of water and boil twice, each time for 1 hour, combine the decoctions, filter, concentrate the filtrate into a thick paste, dry under reduced pressure into a dry paste powder, crush, and prepare a solution of corresponding concentration before use.

[0047] Pharmacodynamic evaluation:

[0048] Experimental animals: Male SD rats, 200-220g. 48 male SD rats were weighed and randomly divided into blank group, model group, Y1 low-dose group (7.45g / kg, raw drug amount / body weight), Y1 high-dose group (14.9g / kg, raw drug amount / body weight), Y2 group (10.2g / kg, raw drug amount / body weight), polyene phosphatidylcholine group (0.1425g / kg), 8 rats in each group, 4 rats in each cage, the model group and the drug group were fed with an equal amount of methionine and choline-deficient feed (MCD feed) per cage every day, and the blank group was fed with an equal amount of MCS control feed per cage every day. After feeding for 2 weeks, the drug group was given the corresponding drug treatment according to the above dose, and the blank group and the model group were given the corresponding volume of distilled water by gavage, and the drug administration period was 8 weeks.

[0049] Detection of liver weight and liver index: After 8 weeks of administration, the livers of rats were collected and the body weight and liver weight of rats were measured. Liver index = liver weight / body weight × 100%.

[0050] Detection of serum biochemical indices: After 8 weeks of administration, blood was collected from the abdominal aorta, and the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in serum were measured using a fully automatic biochemical analyzer.

[0051] Detection of liver TG: Take the largest lobe of the liver, accurately weigh 100 mg, cut it into pieces, put it in a 2 mL centrifuge tube, add 9 times the volume of anhydrous ethanol at a ratio of weight (g) to volume (mL) of 1:9, mechanically homogenize in an ice water bath, centrifuge at 2500r / min for 10 minutes, take the supernatant, and measure it using a triglyceride (TG) kit.

[0052] Male SD rats were anesthetized and liver samples were obtained. The largest lobe of the liver was fixed in 4% paraformaldehyde for 48 hours, paraffin sections and frozen sections were prepared, and liver HE staining, Masson staining and Oil Red O staining were performed for liver histopathological analysis. The NASH pathological diagnosis criteria were evaluated using the NAFLD activity score (NAS).

[0053] Effects on inflammatory factors and liver fibrosis indicators in rats:

[0054] Take liver tissue, weigh about 100 mg, cut it into pieces, put it in a 2 mL enzyme-free centrifuge tube, add 1 mL of trisphenol (Trizol) reagent, extract total RNA from the liver of each rat according to the Trizol method, use ultra-micro spectrophotometer to detect the concentration and purity of RNA, and use EvoM-MLVRT reverse transcription kit (reaction conditions: 1 μg Total RNA, 37 ° C, reverse transcription reaction 15 min, 85 ° C, reverse transcriptase inactivation 5 s, 4 ° C, ∞) to synthesize RNA into cDNA and store it in a -80 ° C refrigerator. Then, use Green Pro Taq HS premixed qPCR kit was used for PCR amplification (step 1: 95°C, pre-denaturation for 30 s; step 2: 95°C, denaturation for 5 s, 60°C, annealing for 30 s, 40 cycles; step 3: dissociation phase). GAPDH was used as the internal reference gene, and the expression level of the target gene was calculated using the 2-ΔΔCt method. The primer sequences are shown in Table 1.

[0055] Table 1:

[0056]

[0057] Effects on liver weight and liver index in rats with non-alcoholic steatohepatitis:

[0058] Compared with the blank group, the liver weight and liver index of the rats in the model group were significantly increased (P<0.001), increasing by about 50.79% and 67.98%, respectively; compared with the model group, the liver weight and liver index of the rats in the polyene phosphatidylcholine group, Y1 low-dose group, Y1 high-dose group and Y2 group were significantly decreased (P<0.05), the liver weight decreased by about 21.35%, 21.59%, 17.53% and 23.58%, respectively, and the liver index decreased by about 17.94%, 14.31%, 15.50% and 16.23%, respectively, as shown in Table 2.

[0059] Table 2:

[0060] Grouping Dosage (g / kg) Liver weight (g) Liver index (%) Blank Group -- 10.00±0.64 2.44±0.08 Model Group -- <![CDATA[15.07±1.73 ### ]]> <![CDATA[4.11±0.28 ### ]]> Polyene phosphatidylcholine group 0.1425 <![CDATA[11.86±0.54 ** ]]> <![CDATA[3.37±0.15 *** ]]> Y1 low dose group 7.45 <![CDATA[11.82±2.02 * ]]> <![CDATA[3.52±0.32 * ]]> Y1 high dose group 14.9 <![CDATA[12.43±0.84 ** ]]> <![CDATA[3.47±0.16 ** ]]> Y2 Group 10.2 <![CDATA[11.52±1.06 ** ]]> <![CDATA[3.44±0.19 ** ]]>

[0061] Note:" ### " Compared with the blank group, P < 0.001; " * ","** "," *** ” indicates P < 0.05, P < 0.01, and P < 0.001, respectively, compared with the model group.

[0062] Effects on serum biochemical parameters of rats with non-alcoholic fatty liver disease:

[0063] Compared with the blank group, the serum ALT, AST and ALP of the model group rats were significantly increased (P<0.001), which were increased by about 1.75 times, 0.62 times and 0.91 times respectively; after drug intervention, compared with the model group, the ALT, AST and ALP of the polyene phosphatidylcholine group, Y1 low-dose group, Y1 high-dose group and Y2 group were significantly reduced (P<0.05), and the ALT content was reduced by about 53.28%, 44.57%, 58.96% and 40.15% respectively, the AST content was reduced by about 36.58%, 30.46%, 35.97% and 28.90% respectively, and the ALP content was reduced by about 23.51%, 21.60%, 36.90% and 27.38% respectively, as shown in Table 3. The results showed that the Chinese medicine compositions Y1 and Y2 can improve the liver function damage of NASH rats.

[0064] Table 3:

[0065]

[0066] Note:" ### " Compared with the blank group, P < 0.001; " * "," ** "," *** ” indicates P < 0.05, P < 0.01, and P < 0.001, respectively, compared with the model group.

[0067] Effects on liver TG in rats with non-alcoholic steatohepatitis:

[0068] Compared with the blank group, the TG content in the liver of the model group rats increased significantly (P<0.001), increasing by about 1.16 times; after 8 weeks of drug treatment, compared with the model group, the TG content in the polyene phosphatidylcholine group, the low-dose Y1 group, the high-dose Y1 group and the Y2 group decreased significantly (P<0.001), decreasing by about 36.79%, 32.63%, 52.50% and 24.10%, respectively, as shown in Table 4. The results showed that both the Chinese medicine compositions Y1 and Y2 could reduce the lipid accumulation in the liver of NASH rats.

[0069] Table 4:

[0070] Grouping Dosage (g / kg) TG (mmol / gprot) Blank Group -- 6.57±0.89 Model Group -- <![CDATA[14.19±0.81 ### ]]> Polyene phosphatidylcholine group 0.1425 <![CDATA[8.97±0.85 *** ]]> Y1 low dose group 7.45 <![CDATA[9.56±1.2 *** ]]> Y1 high dose group 14.9 <![CDATA[6.75±0.94 *** ]]> Y2 Group 10.2 <![CDATA[10.77±0.66 *** ]]>

[0071] Note:" ### " Compared with the blank group, P < 0.001; "*** ” indicates P < 0.001 compared with the model group.

[0072] Effects on liver pathological tissues in rats with non-alcoholic steatohepatitis:

[0073] like Figure 2 As shown, the liver was stained with HE (Hematoxylin-eosin), and the results showed that the liver lobules of the blank group rats were clear and complete, the hepatocytes were arranged neatly and orderly, the hepatic cords and hepatic sinusoids were arranged radially around the central vein, and there was no fatty degeneration, ballooning and inflammatory cell infiltration; while the basic structure of the liver lobules of the model group rats was destroyed, the hepatocytes were arranged disorderly, a large number of fat vacuoles appeared in the cytoplasm, the nuclei were condensed and aggregated, the fat was severely degenerated, and accompanied by inflammatory infiltration and ballooning; the fat vacuoles in the drug group polyene phosphatidylcholine, low-dose Y1, high-dose Y1 and Y2 groups decreased, the inflammatory infiltration decreased, and the hepatocytes tended to be normal.

[0074] Image J software was used to quantitatively analyze the area of ​​fatty degeneration in the liver by HE staining, and NAS score was performed. Figure 3 As shown, Figure 3 The statistical test method uses the one-way analysis of variance (ANOVA) method to compare data. The P value is a measure of the probability of the observed result occurring in the hypothesis test. The smaller the P value, the higher the significance of the result. ### " Compared with the blank group, P < 0.001; " * "Indicates that compared with the model group, P < 0.05, which means that the results are significantly different;" ** "Indicates that compared with the model group, P < 0.01, which means that the results are highly significant;" *** ” indicates that compared with the model group, P<0.001, which means that the results are extremely significantly different. Figure 3 It can be seen that the scores of liver fatty degeneration, lobular inflammation and ballooning in the model group were significantly higher than those in the blank group, and the NAS comprehensive score was greater than 5, which indicated that the model had progressed to NASH according to the diagnostic criteria of NAFLD; after intervention with polyene phosphatidylcholine, low-dose Y1, high-dose Y1 and Y2 groups, the scores of liver fatty degeneration, lobular inflammation and ballooning and the NAS comprehensive score were significantly reduced.

[0075] like Figures 4-5 As shown, the liver was stained with Oil Red O. Figure 5The statistical test method used was one-way analysis of variance (ANOVA) for data comparison; the results showed that the blank group rats had basically no red lipid droplets, while the model group rats had a large number of red lipid droplets in the liver, and the lipid droplets in the drug group polyene phosphatidylcholine, low-dose Y1, high-dose Y1 and Y2 groups were significantly reduced. Image J software was further used to quantitatively analyze the area of ​​Oil Red O lipid droplet staining (Table 5). The results showed that compared with the model group, the lipid droplet staining area in the polyene phosphatidylcholine, low-dose Y1, high-dose Y1 and Y2 groups was significantly reduced (P<0.001), which indicates that the Chinese medicine compositions Y1 and Y2 can significantly reduce lipid accumulation in NASH rats.

[0076] Table 5:

[0077] Grouping Dosage (g / kg) Lipid droplet staining area Fibrosis area Blank Group -- 0±0％ 0.18±0.07％ Model Group -- <![CDATA[32.98±1.26% ### ]]> <![CDATA[5.15±1.17% ### ]]> Polyene phosphatidylcholine group 0.1425 <![CDATA[10.90±1.67% *** ]]> <![CDATA[1.69±0.25% * ]]> Y1 low dose group 7.45 <![CDATA[10.16±1.01% *** ]]> <![CDATA[0.64±0.07% ** ]]> Y1 high dose group 14.9 <![CDATA[7.65±1.45% *** ]]> <![CDATA[0.56±0.05% ** ]]> Y2 Group 10.2 <![CDATA[11.32±1.57% *** ]]> <![CDATA[0.80±0.05% ** ]]>

[0078] Note:" ### " Compared with the blank group, P < 0.001; " *** ” indicates P < 0.001 compared with the model group.

[0079] like Figures 6-7 As shown, the liver was stained by Masson staining, where Figure 7 The statistical test method used was single factor analysis of variance (ANOVA) for data comparison; the results showed that the liver tissue of the blank group rats had basically no fibrosis, while the liver tissue of the model group rats had significant fibrosis, with blue-stained collagen fibers centered in the portal area and spreading out in a star shape, with multiple areas connected to form a "bridge"; the liver tissue fibrosis of the drug group Y1 low-dose group, Y1 high-dose group, and Y2 group was significantly reduced, with only a small amount of blue-stained collagen fibers in the portal area, and no "bridge" structure was observed. The Masson staining area was further quantitatively analyzed using Image J software (Table 5). The results showed that compared with the model group, the fibrosis area of ​​the polyene phosphatidylcholine group (P<0.05), Y1 low-dose, Y1 high-dose, and Y2 groups was significantly reduced (P<0.01), indicating that polyene phosphatidylcholine, Chinese medicine composition Y1, and Y2 can significantly inhibit liver fibrosis in NASH rats, and the effects of Y1 and Y2 are better than polyene phosphatidylcholine.

[0080] Effects on inflammatory factors in the liver of rats with non-alcoholic steatohepatitis:

[0081] Compared with the blank group, the mRNA expression levels of inflammatory factors TNF-α, IL-6, IL-1β and MCP-1 in the model group were significantly increased (P<0.001). Compared with the model group, the mRNA expression levels of TNF-α, IL-6, IL-1β and MCP-1 in the drug group polyene phosphatidylcholine group, Y1 low-dose group, Y1 high-dose group and Y2 group were significantly decreased (P<0.01). Figure 8 This indicates that both the Chinese medicine compositions Y1 and Y2 can down-regulate the expression of inflammatory factors in NASH rats.

[0082] Effects of scleroderma on liver fibrosis factors in rats with nonalcoholic steatohepatitis

[0083] Compared with the blank group, the mRNA expression levels of profibrotic factors TGFβ, Acta2, Col1a1, and Col1a2 in the model group were significantly increased (P<0.001); compared with the model group, the mRNA expression levels of TGFβ in the polyene phosphatidylcholine group, Y1 low-dose group, Y1 high-dose group, and Y2 group were significantly decreased (P<0.01). Fig. 9 It shows that polyenylphosphatidylcholine, Chinese medicine composition Y1, and Y2 can significantly downregulate the expression of fibrosis factors in NASH rats, and some indicators of Y1 and Y2 groups are better than those of polyenylphosphatidylcholine group.

[0084] like Figure 1 As shown, based on the above-mentioned multi-faceted index evaluation results, it can be seen that the pharmaceutical composition of the present invention can improve liver function damage, liver lipid accumulation, liver fatty degeneration, liver inflammation and fibrosis caused by non-alcoholic fatty hepatitis. The present invention can provide drug options for the treatment of non-alcoholic fatty hepatitis and lay a working foundation for clinical drug development.

[0085] The above is only an embodiment of the present invention, and the common knowledge such as the known specific technical solutions or characteristics in the solution is not described in detail here. It should be pointed out that for those skilled in the art, several modifications and improvements can be made without departing from the technical solution of the present invention, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A pharmaceutical composition for improving non-alcoholic fatty hepatitis, characterized in that: The pharmaceutical composition is composed of the following components in weight ratio: 10-60 parts of bupleurum, 0-30 parts of scutellaria, 0-90 parts of salvia miltiorrhiza, 0-30 parts of cinnamon twig, 0-30 parts of dried ginger, 0-45 parts of hawthorn, 0-45 parts of trichosanthes, 0-90 parts of raw oyster, and 0-20 parts of roasted liquorice.

2. The pharmaceutical composition for improving non-alcoholic steatohepatitis according to claim 1, characterized in that: The pharmaceutical composition is composed of the following components in weight ratio: 20-60 parts of bupleurum, 10-30 parts of scutellaria, 30-90 parts of salvia miltiorrhiza, 10-30 parts of cinnamon twig, 10-30 parts of dried ginger, 15-45 parts of hawthorn, 15-45 parts of trichosanthes, 30-90 parts of raw oysters, and 10-20 parts of roasted liquorice.

3. A pharmaceutical preparation for improving non-alcoholic fatty hepatitis, characterized in that: The active ingredient of the pharmaceutical preparation is the pharmaceutical composition according to claim 1 or 2.

4. The pharmaceutical preparation for improving non-alcoholic steatohepatitis according to claim 3, characterized in that: The pharmaceutical preparation is prepared by mixing the pharmaceutical composition in proportion, extracting the mixture, and utilizing the extract.

5. Use of the pharmaceutical preparation according to any one of claims 3 to 4 in improving diseases or symptoms related to non-alcoholic fatty liver disease.

6. The use according to claim 5, characterized in that: Symptoms of nonalcoholic steatohepatitis include hepatic steatosis, hepatic inflammation and fibrosis, liver function damage, and hepatic lipid accumulation.

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