CN110772547A - Application of Wenwangyibi extract in preparing medicine for treating hepatitis - Google Patents
Application of Wenwangyibi extract in preparing medicine for treating hepatitis Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A61K2236/30—Extraction of the material
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- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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Abstract
The invention discloses an application of a radix seu caulis Wednoniae extract in preparing a medicament for treating hepatitis, wherein a total extract, a water extraction and alcohol precipitation part and an ethyl acetate part which are obtained by decocting the radix seu caulis Wednoniae with water can obviously inhibit the secretion of inflammatory factors TNF- α and IL-1 β stimulated by PA, can inhibit the expression of mRNA of inflammatory factors TNF- α, IL-1 β and the like in AML12 cells in vitro, and can obviously inhibit the expression of mRNA of inflammatory factors TNF- α, IL-1 β and the like in mouse liver tissues induced by HFD in vivo.
Description
Technical Field
The invention relates to the field of medicines, and in particular relates to an application of a Wenwangzhipen extract in preparing a medicine for treating hepatitis.
Background
Liver is an organ which is often attacked by various pathogenic factors or diseases, and liver injury can cause metabolic disorders of various substances, resulting in disorder of various systems and diseases related thereto. Inflammatory liver injury is a common mechanism for the occurrence and development of various acute and chronic liver diseases, and the main pathological manifestations are liver inflammatory cell infiltration, increased expression of inflammatory factors, and massive necrosis and apoptosis of liver parenchymal cells. Inflammatory liver injury is usually an early manifestation of various liver diseases, and can further progress into hepatic fibrosis, liver cirrhosis and even liver cancer, and proper intervention and treatment at the stage are the key points for preventing irreversible lesion of the liver. There are many factors that cause inflammatory liver injury, and the common factors include alcohol, high lipid, virus, drug, autoimmunity, etc. With the improvement of the living standard of substances and the influence of modern life style, the incidence and the number of diseases of non-alcoholic fatty liver diseases and alcoholic liver diseases are increased dramatically, and the liver diseases become a main part of non-infectious liver diseases.
The non-alcoholic fatty liver disease is a clinical pathological syndrome for eliminating the liver lipid metabolism disorder caused by alcohol and other definite liver injury factors, is one of the common chronic liver diseases in China, and causes great medical burden. Nonalcoholic steatohepatitis (NASH) is a key stage in the progression of simple fatty liver to end-stage liver disease, and is characterized mainly by steatosis and inflammatory cell infiltration of liver cells. Western medicine mainly adopts drug therapy in treating NASH, including combined therapy of lipid-lowering, sugar-lowering, liver-protecting drugs, insulin sensitizer, bile acid secretion promoting drugs and anti-oxidation drugs, but clinical effect observation shows that the current clinical drugs have poor therapeutic effect and large side effect, so the research and development of new drugs are not slow.
The alcoholic liver disease is a liver injury disease caused by long-term excessive drinking, the incidence rate of the alcoholic liver disease is increased year by year, the second major liver injury reason after viral liver injury is formed in China, the pathogenesis of the alcoholic liver disease is complex, the toxic effect of ethanol, immune inflammatory reaction, oxidative stress, genetic factors and the like all participate in the pathophysiological process, the alcoholic hepatitis (ASH) is an important stage in the progress of the alcoholic liver disease, can be effectively reversed through the abstinence of alcohol and proper drug intervention and is a key for preventing the alcoholic liver disease from progressing, and although research on the ASH is carried out for decades, the drugs for treating the ASH, such as glucocorticoid, phosphodiesterase inhibitor, TNF- α antibody and the like, still have different problems of great side effect, poor curative effect, great dispute and the like in clinical treatment.
The traditional Chinese medicine generally has the action mechanism of multiple ways and multiple targets, and the advantage makes the traditional Chinese medicine become one of the directions for researching and developing the medicine for treating hepatitis. Because NASH and ASH share many common points in pathogenesis, traditional Chinese medicine mainly treats the two types of hepatitis by clearing heat and removing toxicity, soothing liver and regulating qi, and strengthening spleen and eliminating dampness. Commonly used prescriptions include Yinchengao decoction, heat-clearing, dampness-draining and swelling-reducing decoction, modified by Bupleurum root liver-soothing powder and phrenic stasis-removing decoction, etc.; the single medicine comprises bupleurum, salvia, peach kernel, pseudo-ginseng, scutellaria, forsythia, giant knotweed, oriental wormwood and the like, and the mechanism of the single medicine mainly relates to the aspects of regulating lipid metabolism, inhibiting oxidative stress reaction, improving intestinal barrier, improving inflammatory state and the like. The curative effect analysis of the traditional Chinese medicine for treating the hepatitis shows that the traditional Chinese medicine has definite curative effect on reducing the traditional Chinese medicine clinical syndrome integral of the patient. However, because the traditional Chinese medicine has defects in various aspects such as extraction process, optimal effective dose of compound medicine, complex metabolism in vivo, specific targeting mechanism of the medicine and the like, the standard traditional Chinese medicine for treating NASH and ASH does not exist at present.
The WENWANGYI Pen is a whole plant of Balanophora henryi hemsl (Balanophora henyi hemsl) and its similar species in Balanophora Henryi hemsl of Balanophoraceae, has effects of tonifying liver and kidney, clearing heat and detoxicating, relieving swelling and pain, etc., and is mainly used for treating diseases such as digestive tract hemorrhage, impotence, etc.
Disclosure of Invention
The invention provides an application of a Wenwang Yinbu extract in preparing a medicament for treating hepatitis, and the Wenwang Yinbu extract has a good protection effect on non-alcoholic steatohepatitis and alcoholic steatohepatitis.
The invention provides an application of a Wenwangzhipen extract in preparing a medicament for treating hepatitis.
Furthermore, the hepatitis is nonalcoholic steatohepatitis and alcoholic steatohepatitis.
Further, the extract of the radix astragali Sinici is a total extract (Extzw) obtained by decocting the radix astragali Sinici with water.
Further, the extract of the Strychnos Wenzhnensis is an extract (Frp) of a water extraction and alcohol precipitation part of the Strychnos Wenzhnensis, and is obtained by adding ethanol into a total extract obtained by decocting the Strychnos Wenzhnensis with water, and dissolving the precipitate obtained after the ethanol treatment with water again.
Further, the extract of radix seu herba Wenzenbachiae is an ethyl acetate part extract (Fre) of radix seu herba Wenzenbachiae, which is obtained by adding ethanol into a total extract obtained by decocting with water, collecting the supernatant after ethanol treatment, recovering ethanol, adding appropriate amount of water, adding ethyl acetate for extraction, and evaporating the extract to dryness.
Further, the extract of the King pen is an extract of a water extraction and alcohol precipitation part, and the hepatitis is non-alcoholic steatohepatitis and/or alcoholic steatohepatitis.
The invention also relates to a medicament for treating non-alcoholic steatohepatitis and/or alcoholic steatohepatitis, wherein the effective component of the medicament is the extract of the King Wen Yinbao.
Furthermore, the content of the extract of the Chinese king-appliance crayon in the medicine is 0.1-100 wt%.
The invention searches the existing reports related to the invention, and the inventor finds that the soup spring, the Zhouyanghua and the like report the research on the sobering action mechanism of the extract of the Tupistra chinensis and the Balanophora chinensis, and prepares an animal model by a method of intragastric perfusion of the commercially available white spirit with different concentrations, and finds that the Balanophora chinensis has the effects of relieving alcoholism and resisting alcoholism, and can protect the liver injury caused by alcohol. Compared with the research, the invention adopts the international standardization Lieber-De Carli liquid diet method (chrono-plus-binding improved) to prepare the Chronic ASH animal model, and the result shows that the extract of the king pen has good protective effect on alcoholic hepatitis, and the mechanism of the extract is related to that the extract of the king pen obviously enhances the intestinal shielding function, reduces the endotoxin entering the liver from the intestinal and interferes the intestinal-hepatic axis. Different from the previous reports, the finding shows that the extract of the Wenwangyibi pen plays a role in protecting ASH by protecting the intestinal tract shielding function damage induced by alcohol.
The biological activity of the reported King pen related to the invention is searched, and no report about the effect of the Balanophora stolonifera drug on the nonalcoholic steatohepatitis is found. The cause of non-alcoholic steatohepatitis is not well defined compared to alcoholic hepatitis. Several studies have shown that NASH occurs in association with various factors such as insulin resistance, genetic susceptibility, immunity, and metabolic disorders. The results of animal models prepared by high-fat diet feeding and in-vitro cell tests show that the King-Yinbao extract obviously reduces the inflammatory reaction of the liver in NASH and has good protection effect on NASH.
On the basis of confirming the effect of the water extract of the Wenwang-Yizhu in the treatment of NASH and ASH, different extracts are further obtained by methods of alcohol precipitation, ethyl acetate extraction and the like so as to screen, compare and confirm the effective parts of the extracts.
The inventor finds in research that various extracts obtained from the King pencils can remarkably inhibit the expression of inflammatory factor mRNA and protein of palmitic acid stimulated liver AML12 cells in vitro and remarkably reduce the expression of inflammatory factor mRNA in liver tissues in a high-fat diet induced non-alcoholic steatohepatitis mouse; simultaneously can inhibit lipid deposition of liver cells of mice with alcoholic hepatitis, resist inflammatory reaction, and has the effects of reducing enzyme and protecting liver. The extract obtained from Wenwang Stringbush can effectively inhibit the inflammatory reaction of liver in non-alcoholic steatohepatitis and alcoholic hepatitis, and has liver protection effect.
Through further research, the extracts of the pen of Shangwang, especially the total extract (Extzw), the water extraction and alcohol precipitation part (Frp) and the ethyl acetate part (Fre) prepared from the water extract can obviously inhibit the secretion of inflammatory factors TNF- α and IL-1 β stimulated by PA, the expression of inflammatory factor mRNA such as TNF- α and IL-1 β and the like in AML12 cells in vitro, and can obviously inhibit the expression of inflammatory factor mRNA such as TNF- α and IL-1 7 in mouse liver tissues induced by HFD in vivo.
Drawings
FIG. 1 is a high performance liquid chromatography characteristic spectrum of the extraction parts of the King pen of the embodiment, namely, the Extrz, Frp and Fre at 245nm and 210nm respectively.
Fig. 2 shows the effect of the extraction sites extrrz, Frp and Fre of wakame on AML12 cell growth in the example (xp <0.05, n ═ 3).
FIG. 3 shows the effect of the extraction sites Extrz, Frp and Fre of King pen on the secretion of TNF- α and IL-1 β from AML12 cells induced by PA (# p <0.05, compounded with Mock group; # p <0.05, compounded with PAgroup).
FIG. 4 shows the effect of the extract sites of Extrz, Frp and Fre of King-pen on the induction of AML12 cytokine mRNA expression by PA in the examples.
FIG. 5 shows the effect of the extract sites of Extrz, Frp and Fre of King-Wen-Stroke on the expression of mRNA for HFD-induced liver tissue inflammatory factor in mice
FIG. 6 is a graph showing the staining of paraffin sections HE of mouse liver tissues in examples. Wherein A is PF: a diet control group; b is AF: an alcoholic hepatitis model group; c is AF + Frp: drug + alcoholic hepatitis model group; d is PF + Frp: drug control group.
FIG. 7 is an oil red staining chart of paraffin sections of liver tissues of mice in the examples. Wherein A is PF: a diet control group; b is AF: c is an alcoholic hepatitis model group; AF + Frp: drug + alcoholic hepatitis model group; d is PF + Frp: drug control group.
FIG. 8 shows the serum ALT and AST activities of the mice in the examples. A, serum ALT activity; b, serum AST activity. PF: a diet control group; AF: an alcoholic hepatitis model group; AF + Frp: drug + alcoholic hepatitis model group; PF + Frp: drug control group. P < 0.05; p < 0.01.
FIG. 9 shows the mRNA expression of the liver tissue TNF- α -1 β -6 and NLRP3 detected by ELISA, B.RT-PCR and the content of TNF- α -1 β in the liver tissue.
FIG. 10 is a paraffin section HE staining of mouse colon tissue in example A; B. a fluorescence staining pattern of zonulin ZO-1 and Occludin of a colon tissue of a mouse; WB detecting the expression of zonulin ZO-1 and Occludin in colon tissue of a mouse; ELISA to detect serum LPS concentration. PF: a diet control group; AF: an alcoholic hepatitis model group; AF + Frp: drug + alcoholic hepatitis model group; PF + Frp: drug control group. P < 0.05; p < 0.01.
FIG. 11 shows the effect of Frp extracted from Wen king crayon on the expression of alcohol-induced human colon cancer cells Caco-2 claudin ZO-1 and Occludin. A. Carrying out immunofluorescence detection on the expression of the human colon cancer cell Caco-2 tight junction protein ZO-1; B. performing immunofluorescence detection on the expression of human colon cancer cell Caco-2 tight junction protein Occludin; RT-PCR detects the expression of Caco-2 tight junction protein ZO-1 and Occludin in human colon cancer cells. PF: a diet control group; AF: an alcoholic hepatitis model group; AF + Frp: drug + alcoholic hepatitis model group; PF + Frp: drug control group. P < 0.05; p < 0.01.
Detailed Description
The invention will be further elucidated with reference to the following examples.
Example (b):
1. sources of drugs
The Wenwang Yinbaibi medicine adopted by the research of the invention is collected in the mountain county of the temple in Shi Wei of Hubei province and is identified as the whole herb of the wild Balanophora Involurata of the mythimna rufii.
2. Extraction and preparation of medicine
Collecting whole plant of Wenwangxiangyinbei, drying, and pulverizing. Decocting with 8 times of water for 3 times, mixing decoctions, and concentrating under reduced pressure to obtain medicinal liquid with density of about 1.10 to obtain total extract (Extzw); adding ethanol into the liquid medicine under the condition of 55 ℃ heat preservation and stirring until the alcohol content is 70%; standing at low temperature overnight (12h), and centrifuging to separate out precipitate; washing the precipitate with appropriate amount of ethanol for several times, volatilizing ethanol, adding appropriate amount of water to dissolve the precipitate again to obtain water extraction and ethanol precipitation part (Frp); recovering ethanol from the supernatant, adding appropriate amount of water, extracting with equal volume of ethyl acetate for 3 times, mixing extractive solutions, and concentrating to dry to obtain ethyl acetate fraction (Fre).
3. Characteristic spectrogram of each extracted part of Wenwang pen
HPLC characteristic maps of the total extract (Extzw), the ethyl acetate part (Fre), the water extraction and alcohol precipitation part (Frp) and the like prepared from the water extract of the medicinal material of the Wenwangyi Diels are researched. The column used octadecylsilane chemically bonded silica as a packing material (250X 4.6mm, particle size 5 μm). The mobile phase system uses acetonitrile as a mobile phase A and water as a mobile phase B to carry out gradient elution, wherein the elution procedure is 0min and 5% of A is adopted; 10min, 15% A; 35min, 25% A; 50min, 30% A; 70min, 65% A. The column temperature was 30 ℃, the flow rate was 1.0ml/min, and the detection wavelengths were 254nm and 210 nm.
4. Material and method for researching NASH and ASH protection effect of each extracted part of Wenwang-Yinbao
4.1 cell culture and experimental animals, reagents:
AML12 cells were purchased from cell resource center of Shanghai Life sciences research institute of Chinese academy of sciences, cultured in DMEM containing 10% fetal calf serum, and placed at 37 deg.C and CO
2The culture was carried out in an incubator with a volume fraction of 5%. DMEM medium and fetal bovine serum were purchased from Gibco, USA; kunming mice were purchased from the Experimental animals center, university of three gorges.
4.2 cell proliferation assay:
AML12 cells were seeded in 96-well plates (1X 10)
4One/hole), after 24 hours of cell adherence, adding different mass fractions of ethyl acetate part (Fre), water extraction and alcohol precipitation part (Frp), total extract (Extzw) and the like of the Wenwangzhi pen extract respectively, wherein the mass fraction ranges are 25 mugmL-150. mu.g/mL. Then, the absorbance at 570nm (A570) was measured by the MTT method (5g/L MTT) on days 1, 2, 3 and 4 for each extracted part, and the experiment was repeated 3 times.
4.3 inflammatory cytokine secretion assay:
AML12 cells were seeded in 96-well plates (1X 10)
4One/well), after 24 hours of cell adherence, 50 mug/mL of ethyl acetate part (Fre) of Wenwangyi pen, water extraction and alcohol precipitation part (Frp), total extract (Extzw) and 250 mug M Palmitic Acid (PA) are respectively added, a zero adjusting hole, a reagent control group (Mock), a PA group, PA + extracts (Fre, Frp and Extzw) of Wenwang pen are arranged, each group has 4 holes, and after 24 hours of drug action, the secretion of TNF- α and IL-1 β of supernatant is detected by an ELISA kit.
4.4RT-PCR detection of cellular level expression of mRNA for the relevant inflammatory factor:
AML12 cell suspension was seeded in 6-well plates (2X 10)
5One/hole), after the cells adhere to the wall, 50 mu g/mL of Wangbaopen extract ethyl acetate part (Fre), water extraction and alcohol precipitation part (Frp), total extract (Extzw) and the like are respectively added to act on the cells for 12 hours, reagent control (Mock) groups, PA groups and PA + Wenwaopen extract groups are set, 250 mu M PA is added to jointly stimulate for 12 hours, total RNA is extracted according to a Trizol kit, reverse transcription is carried out to obtain cDNA, and then the expression of relevant inflammatory factor mRNA such as TNF- α, IL-1 β and the like is amplified by PCR.
4.5RT-PCR detection of expression of relevant inflammatory factor mRNA in mouse liver tissue:
the method comprises the following steps of providing 6-week-old male Kunming mice (20-23g) from an animal center of university of three gorges, approving experimental animal operation by ethical committee of the university of three gorges, dividing Kunming mice into a reagent control (Mock) group, a High Fat Diet (HFD) feeding group, an ethyl acetate part (Fre), a total extract (Extzw) and a water extraction and alcohol precipitation part (Frp) group after the Kunming mice are adaptively fed for 1 week, wherein each group of the mice contains 6 mice, each group of the mice freely drink water, the Mock group contains common feed, the Mock group contains High Fat Diet (HFD) feeding for 4 months, the Mock group contains a corresponding part extract of the King pen, and is perfused once a day for four months, the doses are 400mg/kg of Fre, 1000 mg/Frz of the express, 400 mg/kg. of the p of the Mock group and the HFD group contain the same solvent, the mRNA is extracted according to the same volume of the Mock and the mRNA is expressed by a PCR reagent kit, the PCR amplification factor is α, and the total inflammation is extracted after the Mock group is 366332.
4.6 preparation of animal model of alcoholic hepatitis:
male C57BL/6 mice (20-25g) 8 to 10 weeks old were provided by the animal center, university of three gorges; laboratory animal procedures were approved by the university of three gorges laboratory animal ethics committee. The C57BL/6 mice were randomly divided into diet control group, alcoholic diet model group, alcoholic diet + water extraction and alcohol precipitation part (400mg/kg), diet control + water extraction and alcohol precipitation part (400 mg/kg). An animal model of chronic alcoholic hepatitis is prepared by adopting a "chronic and binge ethanol feeding" mode. Lieber-Decalic liquid feed (Nantong Tellophile feed science and technology Co., Ltd., TP4030D) was adapted for 5 days. Starting on day 6, model mice were fed with Lieber-DeCarlic liquid diet containing 5% alcohol (V/V) (Nantong Telofu feed science Co., Ltd., TP4030C), and the control group remained unchanged. The mice in the diet control group and the alcoholic diet model group are fed in an isocaloric pairing way. During feeding period, the model group was directly gavaged with alcohol (5g/kg)1 to 2 times per week, while the control group was gavaged with 45% maltodextrin (9g/kg), and fed for 6 weeks. The drug treatment group is used for intragastric administration once a day at a fixed time until the end of molding while feeding the alcohol liquid feed. When the molding of the mice is finished, the mice of the model group are subjected to alcohol intragastric administration at one time, the mice of the control group are subjected to intragastric administration with 45% maltodextrin, the animals are sacrificed after 9 hours, serum is left, and liver tissues are detected.
4.7 mouse serum ALT, AST Activity and liver tissue-associated inflammatory factor expression assay:
the serum ALT and AST activity (Nanjing institute for bioengineering) and liver tissue homogenate TNF- α and IL-1 β content (Invitrogen) of the mice are determined according to the instruction of a kit.
4.8 detection of human colon carcinoma Cacao cell tight junction protein expression:
cacao cells are inoculated on a 6-well plate to prepare a Cacao cell slide, after the cells grow to form a film-forming structure, alcohol (100 mu mol/L) is added for culturing for 24 hours to induce cell damage, a drug treatment group is simultaneously added with 100 mu g/ml of King Wen's crabapple extract water extraction and alcohol precipitation part (Frp), the cell slide is taken out, and the expressions of cell tight junction proteins ZO-1 and Occludin are detected by an immunofluorescence method and RT-PCR.
4.9 statistical treatment
The experimental results are expressed as mean. + -. standard deviation
Shows, analysis using statistical software SPSS11.5, One-Way analysis of variance (One Way ANOVA) between groups, in p<A difference of 0.05 was significant.
5. Results of the study
5.1 extracting the characteristic maps of the parts Extrz, Frp and Fre by one pen of Wen king: the results are shown in FIG. 1.
5.2 Effect of the extract fractions of Extrz, Frp and Fre of King-Yinbao on the growth of AML12 cells
As shown in FIG. 2, in AML12 cells cultured in vitro, when the cells were treated with 100. mu.g/mL of the extract for 4 days and 150. mu.g/mL of the extract for 3 days, respectively, there was a certain reduction in cell growth (p <0.05) compared to the blank control group.
The above results show that: the extraction sites of the King pen, namely, Extrz, Frp and Fre, have no obvious influence on the growth of AML12 cells within the concentration range of less than 100 mu g/mL.
5.3 study of anti-inflammatory action of various extraction parts of Wenwang Yinbao in AML12 cells in vitro
As shown in the attached figures 3-4, after the water extraction and alcohol precipitation part (Frp), the total extract (Extzw) and the ethyl acetate part (Fre) in the extract of the Meretrix gracilistylus are respectively treated on AML12 cells, secretion of inflammatory factors TNF- α, IL-1 β and the like induced by PA (figure 3) can be remarkably inhibited, and the expression of mRNA (figure 4) of inflammatory related factors TNF- α, IL-1 β and the like in AML12 cells can be remarkably inhibited.
5.4 anti-inflammatory action research of various extraction parts of Wenwang Stringbush in vivo
The stomach of a mouse fed with the high-fat feed is perfused by using each extraction part of a pen of Wenwang for four months, as shown in figure 5, the water extraction and alcohol precipitation part (Frp), the ethyl acetate part (Fre) and the total extract (Extzw) can inhibit the expression of part of related inflammatory factors to a certain extent, and the Frp and Fre effects are more obvious, which shows that the Frp, Fre and Extzw in the extraction part of the pen of Wenwang have the function of resisting the high-fat diet induced inflammatory reaction in the mouse.
5.5 study of the protective action of the part extracted by water and precipitated by alcohol of Wenwangyi pen on alcoholic hepatitis
As shown in the attached figures 6-7, compared with a diet control group (PF), when the mice are fed with the alcoholic liquid feed for 6 weeks (AF), the liver of the mice has the phenomena of hepatocyte arrangement disorder, liver cord disappearance, massive lipid deposition in hepatocytes, vacuolation and death, inflammatory cell infiltration and the like, and the successful construction of a mouse model with chronic alcoholic hepatitis is shown. By intragastric administration of a water extraction and alcohol precipitation part (Frp) of the King pen, lipid deposition in liver cells of mice is obviously less, necrosis phenomenon is obviously relieved, and inflammatory cell infiltration is reduced. The result shows that the water extraction and alcohol precipitation part (Frp) of the Wenwangyi pen has a protection effect on liver cells of alcoholic hepatitis.
As shown in figure 8, the water extraction and alcohol precipitation part (Frp) of the Wenwang Stringbush has the effect of significantly reducing the activity of ALT and AST in the serum of the chronic alcoholic hepatitis mouse (p is less than 0.05), and the water extraction and alcohol precipitation part (Frp) of the Wenwang Stringbush has the effect of reducing the enzyme and protecting the liver of the chronic alcoholic hepatitis mouse.
5.6 extraction and alcohol precipitation of the part of Wenwang Stringbush to relieve the liver inflammation reaction of the mice with alcoholic hepatitis
As shown in figure 9, the water extraction and alcohol precipitation part (Frp) of the king crayon can obviously inhibit the expression of TNF- α -1 β -6 and NLRP3 in liver tissues of alcoholic hepatitis (figure 9), and the water extraction and alcohol precipitation part (Frp) of the king crayon is primarily described to reduce the liver inflammatory reaction of an alcoholic hepatitis mouse and play a role in liver protection.
The total extract (Extzw), the water extraction and alcohol precipitation part (Frp) and the ethyl acetate part (Fre) obtained by the extraction method have good protection effect on non-alcoholic steatohepatitis and alcoholic steatohepatitis, and can be independently processed or processed with other components to prepare various liver-protecting medicines or health-care products, the dosage form characteristics of the extract belong to any one of medicinal liquor, tincture, suspension, capsules, tablets, pills, dripping pills, powder and injection, and the extract of the Elsholtzia wenshurica is contained in the extract by 0.1-100% by mass.
5.7 the water extraction and alcohol precipitation part of Wenwangyibi plays a role in protecting alcoholic hepatitis by enhancing the intestinal shielding function
Chronic alcohol intake can damage intestinal barrier function, resulting in intestinal flora imbalance, and endotoxin entering liver through intestinal tract is an important cause of liver injury. The "gut-liver axis" plays a vital role in alcoholic hepatitis.
As shown in figure 10, the water extraction and alcohol precipitation part (Frp) of the Wenwang Stringbush is used for gastric lavage to remarkably protect the intestinal tract of an alcoholic hepatitis mouse, improve the expression of zon-1 and Occludin in the intestinal tract tissue, enhance the intestinal tract shielding function, reduce endotoxin entering the liver through the damaged intestinal tract and relieve liver damage.
As shown in the attached figure 11, the water extraction and alcohol precipitation part (Frp) of the king crayon is used for treating human colon cancer Cacao cells, the expression of alcohol-induced zon-1 and Occludin can be remarkably improved, and preliminary in vitro experiments prove that the water extraction and alcohol precipitation part (Frp) of the king crayon can enhance the shielding function of intestinal cells and further protect alcoholic hepatitis.
Claims (8)
1. The application of Wenwangyizhu extract in preparing medicine for treating hepatitis.
2. Use according to claim 1, characterized in that: the hepatitis is non-alcoholic steatohepatitis and alcoholic steatohepatitis.
3. Use according to claim 1, characterized in that: the extract of the Wenwang Stringbush is a total extract obtained by decocting Wenwang Stringbush with water.
4. Use according to claim 1, characterized in that: the extract of the radix astragali Sinici is the extract of the water extraction and alcohol precipitation part of the radix astragali Sinici, and is obtained by adding ethanol into the total extract obtained by water decoction and re-dissolving the precipitate obtained after ethanol treatment with water.
5. Use according to claim 1, characterized in that: the King Yinbao extract is an ethyl acetate part extract of the King Yinbao, and is obtained by adding ethanol into a total extract obtained by decocting the King Yinbao, taking supernatant obtained after ethanol treatment, recovering ethanol, adding water for dissolving, extracting with ethyl acetate, and evaporating an extract to dryness.
6. Use according to claim 1, characterized in that: the Wenwang Yinbu extract is an extract of a water extraction and alcohol precipitation part, and the hepatitis is non-alcoholic steatohepatitis and/or alcoholic steatohepatitis.
7. A medicine for treating non-alcoholic steatohepatitis and/or alcoholic steatohepatitis is characterized in that an effective component in the medicine is an extract of King Wen Ying pen.
8. The medicament of claim 7, wherein: the content of the extract of Wangbianpen in Chinese medicine is 0.1-100 wt%.
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| CN111733137A (en) * | 2020-07-13 | 2020-10-02 | 江南大学 | Method for screening medicine capable of preventing and/or treating alcoholic liver injury |
| CN112156118A (en) * | 2020-09-17 | 2021-01-01 | 三峡大学 | Application of Wenwangyizhu extract in preparation of kidney fibrosis resisting medicine |
| CN115590897A (en) * | 2022-10-31 | 2023-01-13 | 三峡大学（Cn） | Application of Wenwangyizhuyan extract in anti-aging |
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| CN115590897A (en) * | 2022-10-31 | 2023-01-13 | 三峡大学（Cn） | Application of Wenwangyizhuyan extract in anti-aging |
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