Traditional Chinese medicine composition, its extract and application

Abstract

The present invention relates to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition, its extract and application. The present invention provides a traditional Chinese medicine composition, which is made from the following raw materials in parts by weight: 3-30 parts of hawthorn, 1-30 parts of chicken gizzard-membrane, 1-30 parts of American ginseng, 5-50 parts of houttuynia cordata, 3-60 parts of black beans, 5-30 parts of wild jujube seeds, 2-30 parts of allium macrostemon bunge, 5-30 parts of malt, 1-30 parts of seabuckthorn, 1-30 parts of galangal and 1-30 parts of turmeric. The present invention discovers that the traditional Chinese medicine composition has the function of anti-hepatic fibrosis, extracts its active ingredients, develops a drug with the function of regulating the expression of key cytokines in the process of hepatic fibrosis occurrence, and clarifies its action effect at the cellular and molecular levels, and has broad application prospects in the treatment or adjuvant treatment of hepatic fibrosis and cirrhosis diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition, its extract and application. Background Art

[0002] Liver fibrosis is an important pathological stage in the progression of various chronic liver diseases. It is the result of the continuous wound healing response of the liver to various chronic injuries, mainly manifested by the abnormal proliferation and deposition of the extracellular matrix (ECM), accompanied by the necrosis and apoptosis of hepatocytes. Persistent liver injury leads to abnormal liver structure, and the liver parenchyma is replaced by scar tissue, forming liver cirrhosis, and even developing into liver cancer and liver failure. This not only seriously affects the quality of life and prognosis of patients, but also increases the social and economic burden, and is a public health problem that cannot be ignored. At present, the treatment of anti-liver fibrosis in Western medicine is mainly targeted at the treatment of the primary disease cause and anti-inflammatory and liver-protecting treatments, and there is no clear chemical or biological drug for clinical use in anti-liver fibrosis.

[0003] The activation of hepatic stellate cells (HSCs) is the core link in the occurrence and development of liver fibrosis. After the liver is continuously damaged, HSCs are activated and transformed into myofibroblast-like cells. α-Smooth Muscle Actin (α-SMA) is considered to be a specific marker of myofibroblasts. Activated HSCs significantly express α-SMA, and at the same time secrete and release a large number of pro-inflammatory and profibrotic cytokines. For example, profibrotic factors COL3A1, COL1A1, EDN1, FGF7, FGF2, WNT3, MAP3K5; pro-inflammatory factors NFKB1, TLR2; anti-liver fibrosis cytokines NR4A1, BMP6. Therefore, for this chronic and complex disease of liver fibrosis, developing drugs that can regulate the expression of multiple important cytokines involved in the progression of liver fibrosis is of great significance for the treatment and clinical application of liver fibrosis. Summary of the Invention

[0004] In view of this, the present invention provides a traditional Chinese medicine composition, its extract and application. The present invention provides a traditional Chinese medicine composition, which is made from the following raw materials in parts by weight: 3-30 parts of hawthorn, 1-30 parts of chicken gizzard-membrane, 1-30 parts of American ginseng, 5-50 parts of houttuynia cordata, 3-60 parts of black beans, 5-30 parts of wild jujube seeds, 2-30 parts of allium macrostemon bunge, 5-30 parts of malt, 1-30 parts of seabuckthorn, 1-30 parts of galangal and 1-30 parts of turmeric. The present invention discovers a traditional Chinese medicine composition with anti-liver fibrosis function through experiments, extracts its active ingredients, develops a drug that can regulate the expression of key cytokines during the occurrence of liver fibrosis, and clarifies its effect at the cellular and molecular level.

[0005] To achieve the above-mentioned invention object, the present invention provides the following technical solutions:

[0006] The present invention provides a traditional Chinese medicine composition, which is made from raw materials in the following parts by weight:

[0007] 3 - 30 parts of hawthorn, 1 - 30 parts of chicken gizzard membrane, 1 - 30 parts of American ginseng, 5 - 50 parts of houttuynia cordata, 3 - 60 parts of black beans, 5 - 30 parts of wild jujube seeds, 2 - 30 parts of Chinese chives, 5 - 30 parts of malt, 1 - 30 parts of seabuckthorn, 1 - 30 parts of galangal, and 1 - 30 parts of turmeric.

[0008] In some specific embodiments of the present invention, the traditional Chinese medicine composition is made from raw materials in the following parts by weight:

[0009] 30 parts of hawthorn, 20 parts of chicken gizzard membrane, 15 parts of American ginseng, 50 parts of houttuynia cordata, 3 parts of black beans, 5 parts of wild jujube seeds, 2 parts of Chinese chives, 5 parts of malt, 1 part of seabuckthorn, 1 part of galangal, and 1 part of turmeric; or

[0010] 3 parts of hawthorn, 1 part of chicken gizzard membrane, 1 part of American ginseng, 5 parts of houttuynia cordata, 60 parts of black beans, 30 parts of wild jujube seeds, 20 parts of Chinese chives, 5 parts of malt, 1 part of seabuckthorn, 1 part of galangal, and 1 part of turmeric; or

[0011] 3 parts of hawthorn, 1 part of chicken gizzard membrane, 1 part of American ginseng, 5 parts of houttuynia cordata, 3 parts of black beans, 5 parts of wild jujube seeds, 2 parts of Chinese chives, 30 parts of malt, 20 parts of seabuckthorn, 15 parts of galangal, and 20 parts of turmeric; or

[0012] 16.5 parts of hawthorn, 10.5 parts of chicken gizzard membrane, 8 parts of American ginseng, 27.5 parts of houttuynia cordata, 3 parts of black beans, 5 parts of wild jujube seeds, 2 parts of Chinese chives, 5 parts of malt, 1 part of seabuckthorn, 1 part of galangal, and 1 part of turmeric; or

[0013] 3 parts of hawthorn, 1 part of chicken gizzard membrane, 1 part of American ginseng, 5 parts of houttuynia cordata, 31.5 parts of black beans, 17.5 parts of wild jujube seeds, 11 parts of Chinese chives, 5 parts of malt, 1 part of seabuckthorn, 1 part of galangal, and 1 part of turmeric; or

[0014] 3 parts of hawthorn, 1 part of chicken gizzard membrane, 1 part of American ginseng, 5 parts of houttuynia cordata, 3 parts of black beans, 5 parts of wild jujube seeds, 2 parts of Chinese chives, 17.5 parts of malt, 10.5 parts of seabuckthorn, 8 parts of galangal, and 10.5 parts of turmeric; or

[0015] 30 parts of hawthorn, 20 parts of chicken gizzard membrane, 15 parts of American ginseng, 50 parts of houttuynia cordata, 60 parts of black beans, 30 parts of wild jujube seeds, 20 parts of Chinese chives, 5 parts of malt, 1 part of seabuckthorn, 1 part of galangal, and 1 part of turmeric; or

[0016] 3 parts of hawthorn, 1 part of chicken gizzard-membrane, 1 part of American ginseng, 5 parts of houttuynia, 60 parts of black beans, 30 parts of wild jujube seeds, 20 parts of Chinese scallion, 30 parts of malt, 20 parts of seabuckthorn, 15 parts of galangal, and 20 parts of turmeric; or

[0017] 30 parts of hawthorn, 20 parts of chicken gizzard-membrane, 15 parts of American ginseng, 50 parts of houttuynia, 3 parts of black beans, 5 parts of wild jujube seeds, 2 parts of Chinese scallion, 30 parts of malt, 20 parts of seabuckthorn, 15 parts of galangal, and 20 parts of turmeric; or

[0018] 30 parts of hawthorn, 30 parts of chicken gizzard-membrane, 30 parts of American ginseng, 30 parts of houttuynia, 30 parts of black beans, 30 parts of wild jujube seeds, 30 parts of Chinese scallion, 30 parts of malt, 30 parts of seabuckthorn, 30 parts of galangal, and 30 parts of turmeric.

[0019] In some specific embodiments of the present invention, the traditional Chinese medicine composition includes any of the following ratios and is made from the following raw materials in parts by weight:

[0020]

[0021] The present invention also provides an extract of the traditional Chinese medicine composition, which is obtained by crushing the traditional Chinese medicine composition, performing Soxhlet extraction, concentrating, and freeze-drying.

[0022] In some specific embodiments of the present invention, the Soxhlet extraction is carried out using a 90% ethanol aqueous solution with a volume 15 times that of the sample.

[0023] In some specific embodiments of the present invention, the preparation method of the extract includes the following steps:

[0024] (I) Take the traditional Chinese medicine composition and crush it;

[0025] (II) Add an ethanol aqueous solution and perform standard Soxhlet extraction;

[0026] (III) Concentrate the extract;

[0027] (IV) Transfer it to a freeze-dryer and perform freeze-drying under reduced pressure for 24 h to obtain a freeze-dried powder of the extract.

[0028] In some specific embodiments of the present invention, the standard Soxhlet extraction time is 3 h.

[0029] In some specific embodiments of the present invention, adding the ethanol aqueous solution includes adding a 90% ethanol aqueous solution with a volume 15 times that of the sample.

[0030] In some specific embodiments of the present invention, the conditions for concentrating the extract include concentrating for 20 min to a volume of 2 mL.

[0031] The present invention also provides the traditional Chinese medicine composition; or the application of the extract in any of the following items:

[0032] (I), preparing a drug for preventing and / or treating liver fibrosis; and / or

[0033] (II), preparing a drug for preventing and / or treating liver cirrhosis; and / or

[0034] (III), preparing a drug for replenishing qi and nourishing yin; and / or

[0035] (IV), preparing a drug for promoting blood circulation to remove blood stasis; and / or

[0036] (V), preparing a drug for clearing heat and detoxifying; and / or

[0037] (VI), preparing a drug for invigorating the spleen and promoting digestion; and / or

[0038] (VII), preparing a drug for resolving turbidity and reducing lipid; and / or

[0039] (VIII), preparing a product for inhibiting the contraction of type I collagen gel.

[0040] In some specific embodiments of the present invention, the prevention and / or treatment of liver fibrosis includes regulating the expression of key cytokines of liver fibrosis;

[0041] The key cytokines of liver fibrosis include any one or more of TLR2, NR4A1, COL3A1, COL1A1, EDN1, NFKB1, BMP6, FGF7, FGF2, WNT3 or MAP3K5.

[0042] In some specific embodiments of the present invention, the regulation of key cytokines of liver fibrosis includes:

[0043] (I), inhibiting pro-fibrotic factors: any one or more of TLR2, COL3A1, COL1A1, EDN1, NFKB1, FGF7, FGF2, WNT3 or MAP3K5; and / or

[0044] (II), activating anti-fibrotic factors: NR4A1 and / or BMP6.

[0045] In some specific embodiments of the present invention, the application includes:

[0046] (I), inhibiting the activation of HSC; and / or

[0047] (II), reducing the production of extracellular matrix.

[0048] The present invention also provides a drug, which includes any of the following:

[0049] (I), the traditional Chinese medicine composition; and / or

[0050] (II), the extract.

[0051] In some specific embodiments of the present invention, the dosage form of the drug includes any one or more of oral decoctions, tablets or pills.

[0052] In some specific embodiments of the present invention, the administration method of the drug includes oral administration and / or injection.

[0053] The present invention includes, but is not limited to, achieving the following beneficial effects:

[0054] The present invention discovers a traditional Chinese medicine composition with anti-hepatic fibrosis function, extracts its active ingredients, develops a drug with the function of regulating the expression of key cytokines during the occurrence of hepatic fibrosis, and clarifies its action effect at the cellular and molecular levels, showing broad application prospects in the treatment or adjuvant treatment of hepatic fibrosis and cirrhosis diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art.

[0056] Figure 1 Showing the effects of each single herb in the traditional Chinese medicine composition on the expression of key genes of hepatic fibrosis;

[0057] Figure 2 Showing the effect of the traditional Chinese medicine composition on the expression of the COL1A1 gene;

[0058] Figure 3 Showing the effect of the traditional Chinese medicine composition on the expression of the COL3A1 gene;

[0059] Figure 4 Showing the effect of the traditional Chinese medicine composition on the contraction of type I collagen gel;

[0060] Figure 5 Showing the regulation of the traditional Chinese medicine composition with different ratios on the genes related to hepatic fibrosis;

[0061] Figure 6 Showing the regulation of the traditional Chinese medicine composition and after removing the drug on the genes related to hepatic fibrosis. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0062] The present invention discloses a traditional Chinese medicine composition, its extract and application. Those skilled in the art can draw on the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all regarded as included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate changes and combinations to the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0063] Existing technical solution:

[0064] In the traditional Chinese medicine formula, each single herb, including hawthorn, chicken gizzard membrane, American ginseng, houttuynia cordata, black bean, wild jujube seed, Chinese chive, malt, seabuckthorn, galangal, and turmeric, is selected in a certain proportion to prepare an extract, which can significantly regulate the expression of key cytokines TLR2, NR4A1, COL3A1, COL1A1, EDN1, NFKB1, BMP6, FGF7, FGF2, WNT3, or MAP3K5 involved in the occurrence of fibrosis.

[0065] The key point of the existing technical solution lies in the newly discovered effects of the extracts of each single herb in the traditional Chinese medicine formula in anti-fibrosis, as well as their applications in the treatment and adjuvant treatment of liver fibrosis diseases. The functions and indications of the traditional Chinese medicine formula described in the present invention are: replenishing qi and nourishing yin, promoting blood circulation to remove blood stasis, clearing heat and detoxifying, strengthening the spleen and promoting digestion. It is applicable to liver fibrosis and liver cirrhosis, with clinical manifestations such as fatigue, discomfort or pain in the liver area, dark red or dull tongue, varicose veins under the tongue, and stringy and thready pulse. The formula explanation and mechanism of action are as follows:

[0066] This formula replenishes qi and nourishes yin, promotes blood circulation to remove blood stasis, clears heat and detoxifies, strengthens the spleen and promotes digestion, and reduces turbidity and lipid. In the formula, hawthorn promotes digestion and strengthens the stomach, and promotes blood circulation to remove blood stasis; turmeric promotes blood circulation to remove blood stasis. The two herbs together are the monarch herbs of this formula. Seabuckthorn promotes digestion and strengthens the spleen, and promotes blood circulation to remove blood stasis; American ginseng replenishes qi and promotes the production of body fluid; wild jujube seed nourishes the heart and tonifies the liver; malt promotes digestion and strengthens the spleen. The four herbs together are the minister herbs of this formula. Chinese chive promotes qi movement and relieves stagnation; black bean benefits essence and nourishes blood; galangal promotes qi movement and dissipates binds to relieve pain; chicken gizzard membrane promotes digestion and strengthens the stomach; houttuynia cordata clears heat and detoxifies and promotes diuresis. The above herbs together are the assistant herbs. The combination of all herbs achieves the effects of replenishing qi and nourishing yin, promoting blood circulation to remove blood stasis, clearing heat and detoxifying, and strengthening the spleen and promoting digestion.

[0067] Although there are many research formulas of traditional Chinese medicine for anti-liver fibrosis, the research mechanism of the formula's effects is not clearly explained, and the research often focuses on a single herb, with certain limitations, and the overall research is not systematic and in-depth enough.

[0068] The purpose of the present invention:

[0069] To discover a traditional Chinese medicine composition with anti-liver fibrosis function, extract its active ingredients, develop a drug with the ability to regulate the expression of key cytokines during the occurrence of liver fibrosis, and clarify its effect at the cellular and molecular levels, with the expectation of applying it to the treatment or adjuvant treatment of liver fibrosis and liver cirrhosis diseases.

[0070] Unless otherwise specified, the raw materials and reagents used in the traditional Chinese medicine composition, its extracts, and applications provided by the present invention can be purchased from the market.

[0071] The following further elaborates the present invention in conjunction with examples:

[0072] Example 1 Preparation of the traditional Chinese medicine formula

[0073] 1. A traditional Chinese medicine formula with anti-fibrotic function, which is composed of the following components by weight: 3 - 30 parts of hawthorn, 1 - 30 parts of chicken gizzard-membrane, 1 - 30 parts of American ginseng, 5 - 50 parts of houttuynia cordata, 3 - 60 parts of black beans, 5 - 30 parts of wild jujube seeds, 2 - 30 parts of allium macrostemon bunge, 5 - 30 parts of malt, 1 - 30 parts of seabuckthorn, 1 - 30 parts of galangal, 1 - 30 parts of turmeric.

[0074] By weight, each ratio includes the following components:

[0075]

[0076]

[0077] 2. Preparation method of the traditional Chinese medicine formula: Weigh 8 - 10 g of each of the formula components of hawthorn, chicken gizzard-membrane, American ginseng, houttuynia cordata, black beans, wild jujube seeds, allium macrostemon bunge, malt, seabuckthorn, galangal, and turmeric. After pulverizing and combining them evenly, add 15 times the volume of 90% ethanol aqueous solution, and perform standard Soxhlet extraction for 3 h. The extract is concentrated at 60 °C for 20 min to a volume of 2 mL, transferred to a freeze dryer for freeze-drying under reduced pressure for 24 h to obtain the freeze-dried powder of the extract. Weigh a certain mass of the freeze-dried powder, dissolve it in DMSO to prepare the mother liquor of the extract, and store it at -20 °C.

[0078] Example 2 Utilize HT 3 S (High-Throughput Targeted Transcriptome Screening, HT 3 S) technology to detect the effects of each single-herb extract on the expression of TLR2, NR4A1, COL3A1, COL1A1, EDN1, NFKB1, BMP6, FGF7, FGF2, WNT3, and MAP3K5 genes based on LX-2 cells

[0079] The gene introduction is shown in Table 1. The specific experimental process is as follows:

[0080] 1. Seed LX-2 cells at 2500 cells / well in a 384-well plate. After 24 h of cell attachment and growth, add 0.8 μL of the traditional Chinese medicine extract with a final concentration of 100 μg / mL to the drug treatment wells, and add the corresponding volume of solvent to the blank control group. Each group is set with 3 replicates. After incubation at 37 °C for 24 h, the cells are lysed.

[0081] 2. Use a high-throughput pipetting platform to perform RNA annealing reaction in the cell lysate to bind the probe to mRNA. Obtain the corresponding mRNA by magnetic bead capture method, use T4 ligase to connect the paired probes complementary to mRNA (probe sequences are shown in Table 2), perform PCR amplification and library construction after elution, and finally complete sequencing on the Illumina Xten platform.

[0082] 3. Data processing: The R package DESeq2 was used to calculate the P-values and fold change values (Fold Change, FC) of genes. The log2 logarithm of the FC value was taken. Genes with |log2FC| > 0.58 and P < 0.05 were considered differentially expressed genes. When log2FC > 0, it indicates upregulation of gene expression, and when log2FC < 0, it indicates downregulation of gene expression. The experimental results are shown in Table 3, Table 4 and Figure 1 .

[0083] Table 1 Research results on the relationship between genes and liver fibrosis

[0084]

[0085]

[0086] Table 2 Probe sequences of liver fibrosis-related genes

[0087]

[0088] Table 3 Regulation of liver fibrosis-related genes by each herb in the traditional Chinese medicine composition (log2FC)

[0089]

[0090] Table 4 Regulation of liver fibrosis-related genes by each herb in the traditional Chinese medicine composition (P-value)

[0091]

[0092]

[0093] The results are as Figure 1 shown in Table 3 and Table 4. In the traditional Chinese medicine composition, Ziziphus jujuba var. spinosa, Houttuynia cordata, Crataegus pinnatifida, and Hordeum vulgare jointly downregulated the expression of the pro-liver fibrosis gene EDN1, and among them, Crataegus pinnatifida had the most significant regulation on this gene; Ziziphus jujuba var. spinosa, Houttuynia cordata, and Crataegus pinnatifida jointly downregulated the expression of the pro-liver fibrosis gene COL3A1, and among them, Ziziphus jujuba var. spinosa had the most significant regulation on this gene; Glycine max, Alpinia officinarum, and Hippophae rhamnoides jointly downregulated the expression of the pro-liver fibrosis gene NFKB1, and among them, Alpinia officinarum had the most significant regulation on this gene; Ziziphus jujuba var. spinosa and Curcuma longa jointly upregulated the expression of the anti-liver fibrosis gene NR4A1, and among them, Curcuma longa had the most significant regulation on this gene; in addition, Gallus gallus domesticus specifically upregulated the expression of the anti-liver fibrosis gene BMP6. Hippophae rhamnoides, Allium macrostemon, Panax quinquefolius, Houttuynia cordata, and Glycine max specifically downregulated the expression of the pro-liver fibrosis and pro-inflammatory genes FGF7, MAPK35, WNT3, FFG2, and TLR2, respectively. The combined use of multiple herbs can inhibit liver fibrosis and inflammatory responses from multiple aspects, and the role of each herb is unique and indispensable.

[0094] Example 3: Inhibitory effect of the traditional Chinese medicine composition extract on the expression of genes promoting liver fibrosis: Using qPCR technology, the effect of the traditional Chinese medicine composition extract on the expression of genes promoting liver fibrosis, namely COL1A1 and COL3A1, was investigated.

[0095] The specific experimental procedure is as follows (taking LX-2 cells as an example):

[0096] 1. LX-2 cells were seeded at a density of 150,000 cells per well in a 12-well plate. After 24 hours of cell attachment and growth, a TGFβ1 inducer with a final concentration of 10 ng / μL and a traditional Chinese medicine composition extract with a ratio of 100 μg / mL were added. Meanwhile, a solvent control group was set up, and the cells were treated for 48 hours.

[0097] 2. RNA was extracted using a kit and reverse-transcribed into cDNA.

[0098] 3. qPCR reaction procedure: 95°C for 3 minutes; (95°C for 3 seconds; 60°C for 30 seconds) for 40 cycles.

[0099] Among them, the primer sequences are as follows:

[0100]

[0101]

[0102] 4. Data processing: Taking the gene expression level of the blank control group as 1, a gene expression level greater than 1 in the drug treatment group indicates upregulation of gene expression, and less than 1 indicates downregulation. (#, p < 0.01 compared with the control group; **, p < 0.01 compared with the induction group; *, p < 0.05 compared with the induction group.) The results of three qPCR experiments are shown in Table 5 and Figures 2 - 3 .

[0103] Table 5 qPCR results of the traditional Chinese medicine composition

[0104]

[0105] The qPCR results showed that the traditional Chinese medicine composition could significantly downregulate the expression of COL1A1 and COL3A1 genes.

[0106] Example 4: Effect of the traditional Chinese medicine composition on the contraction of type I collagen gel: By preparing type I collagen gel, the effect of the traditional Chinese medicine composition extract on the contraction of type I collagen gel was observed.

[0107] The specific experimental procedure is as follows (taking LX-2 cells as an example):

[0108] 1. LX-2 cells were digested with trypsin, centrifuged to discard the supernatant, resuspended in culture medium by pipetting, counted, and the cell suspension was adjusted to 2.0×10 6 / mL;

[0109] 2. Preparation of type I collagen gel: On ice, mix type I rat tail collagen, 1 M NaOH, 10×PBS, and MilliQ according to the dosages shown in Table 6;

[0110] 3. Mix the cell suspension and type I collagen gel quickly at a volume ratio of 1:4. Add the cell-gel mixture to a 12-well plate, 1 mL per well, and incubate in a cell culture incubator at 37 °C for 1 h to allow the collagen to solidify. Then add 1 mL of medium to each well;

[0111] 4. After 24 h, add the inducer TGFβ1 with a final concentration of 10 ng / μL and the extract of the traditional Chinese medicine composition at 400 μg / mL. At the same time, set up a solvent control group. Take vertical photos of each well with a camera at a fixed height every 24 h to record the gel shrinkage, and continuously record for 48 h;

[0112] 5. Data processing: Use Image J software to measure the gel area and calculate the gel shrinkage rate. The calculation formula is: Gel shrinkage rate = [1 - (gel area / gel initial area)] × 100%. (#, p < 0.05 compared with the control group; ##, p < 0.01 compared with the control group; **, p < 0.01 compared with the induction group.) The results of three experiments are shown in Tables 6 - 7 and Figure 4 。

[0113] Table 6 Preparation method of gel solution

[0114]

[0115] Table 7 Effect of traditional Chinese medicine composition on the shrinkage of type I collagen gel

[0116]

[0117] The results of the gel shrinkage experiment showed that compared with the model group, the extract of the traditional Chinese medicine composition could significantly inhibit the shrinkage of type I collagen gel at the 24 h and 48 h time points.

[0118] Example 5 Regulation of traditional Chinese medicine compositions with different ratios on genes related to liver fibrosis: (Taking LX-2 cells as an example)

[0119] Refer to the experimental method in Example 2. According to the experimental results in Table 3, calculate the regulation of the traditional Chinese medicine composition on genes related to liver fibrosis based on 10 ratios. The results are shown in Table 8 and Figure 5 。At 10 ratios, the traditional Chinese medicine composition could regulate genes related to liver fibrosis, and the regulatory effects of ratios 7 - 10 were relatively stronger.

[0120] Table 8 Regulation of each herb in traditional Chinese medicine compositions with different ratios on genes related to liver fibrosis (log2FC)

[0121]

[0122]

[0123] Regulation of liver fibrosis-related genes after the lack of drugs in the traditional Chinese medicine composition in Example 6: (Taking LX-2 cells as an example)

[0124] Referring to the experimental method in Example 2, according to the experimental results in Table 3, the log2FC of the regulation of genes by individual drugs in Formulation 10 was added respectively to calculate the log2FC of the regulation of genes by the entire formula. As shown in Table 9 and Figure 6 As shown, after the sea buckthorn was missing from the traditional Chinese medicine composition, the pro-hepatic fibrosis gene FGF7 that could be inhibited by the original composition of the traditional Chinese medicine composition could not be down-regulated, and at the same time, the inhibitory effect on the pro-hepatic fibrosis NFKB1 gene was weakened; after the houttuynia cordata was missing from the traditional Chinese medicine composition, the inhibitory effects on the pro-hepatic fibrosis genes COL3A1 and EDN1 were weakened, and at the same time, the up-regulating effect on the anti-hepatic fibrosis gene NR4A1 was weakened; after the American ginseng was missing from the traditional Chinese medicine composition, the pro-hepatic fibrosis gene WNT3 that could be inhibited by the original composition of the traditional Chinese medicine composition could not be down-regulated.

[0125] Therefore, the traditional Chinese medicine composition randomly lacking sea buckthorn or (and) houttuynia cordata or (and) American ginseng has a worse regulatory effect on liver fibrosis-related genes than the original traditional Chinese medicine composition (the bold and underlined part), which proves the uniqueness of the role of each drug in the traditional Chinese medicine composition.

[0126] Table 9 Regulation of liver fibrosis-related genes by the traditional Chinese medicine composition and after the lack of drugs (log2FC)

[0127]

[0128]

[0129] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for preventing and / or treating liver fibrosis, characterized in that, it is made from the following raw materials in parts by weight: 30 parts of hawthorn, 30 parts of chicken gizzard-membrane, 30 parts of American ginseng, 30 parts of houttuynia cordata, 30 parts of black beans, 30 parts of wild jujube seeds, 30 parts of allium macrostemon bunge, 30 parts of malt, 30 parts of seabuckthorn, 30 parts of galangal and 30 parts of turmeric.

2. An extract of the traditional Chinese medicine composition according to claim 1, characterized in that, the extract is prepared by crushing the traditional Chinese medicine composition, Soxhlet extraction, concentration and freeze-drying.

3. Use of the traditional Chinese medicine composition according to claim 1 or the extract according to claim 2 in the preparation of a drug for preventing and / or treating liver fibrosis.

4. A drug for preventing and / or treating liver fibrosis, characterized in that, (I), it is made from the traditional Chinese medicine composition according to claim 1; or (II), it is made from the extract according to claim 2.

5. The drug according to claim 4, characterized in that, the dosage form of the drug is an oral preparation or an injection preparation.

6. The drug according to claim 4, characterized in that, the oral preparation is an oral decoction, tablets or pills.

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