| Objective: Control and precaution of liver fibrosis caused by liver injury induced by hepatitis B virus(HBV)play an important role in treating chronic hepatitis B(CHB).Jigucao is derived from the dry herb of Abrus cantoniensis Hance(AC)which belongs to Leguminosae family and Abrus genus species.It is recorded by Chinese Pharmacopoeia,possessing the function of removing-dampness,Removing-aurigo,heat-clearing,detoxification,dispersing stagnated hepatoqi to alleviate pain.It can be used to cure jaundice induced damp-heat,pectoraalgia,gastral cavity swelling pain,and acute mastitis in clinically.Preliminary study have also certificated that AC could treat liver fibrosis.It still exists safe risk for lack of systematic evaluation of chemical compounds and mechanism.Therefore,the project plans to perform a pharmacognostical study and activity evaluation of anti-hepatic fibrosis of AC to provide scientific basis for reasonable exploitation and use of AC.Method:1、Resource distribution and pharmacognostics property of AC were studied by hunting different database,including Flora,Chinese Virtual Heribarium,VIP,Wan-fang database,Pubmed,combining with field survey and market research.2、Paraffin section of stem and leave of AC and AM were used to analysis their difference by microscopic examination.3、The difference of AC and AM were comparative analysis in the level of essential oils and fatty acids.LC-MS analysis of ethyl acetate extracts part and n-butyl alcohol extract part of AC.4、molecular docking and ADMET(Absorption,Distribution,Metabolism,Excretion and Toxicity)echnology were applied to screen active ingredients of 243 chemical compounds of Abrus species and prognosised active constituents class of AC.5、Active compounds screening of anti-hepatic fibrosis of AC using HSC-T6 cell in vitro.6、Active compounds screening of anti-hepatic fibrosis of AC on the level of prevention and treatment through using Kunming mice(KM)to make immuno-hepatic fibrosis model in vivo.Result:1、 Investigation results indicated that AC mainly distributes in Guangxi and Guangdong districts in China,it also distributes in Hunan,Hainan and Xianggang districts in China.It generally grows warmth,wet and drought-tolerant,intolerant to cold and poorly drained.2 、 According to morphological and microscopic identification,we establish characteristic identification key of Abrus cantoniensis Hance and Abrus mollis Hance based on their leaflet,appearance,plant weight,secondary xylem conduit and palisade cell of leaf.It can be used to correctly distinguish them.3、Comparative analysis of essential oils of AC and AM showed that their main common compounds is(±)-α-terpinyl acetate(24.30% and 20.07%).Besides,methyleugenol(22.22%),anethole(14.08%),acetophenol(3.07%),and paeonol(2.41%)of AC is not detected in AM,as α-ethyl-caproic acid is not detected in AC.4、Comparative analysis of fatty acids of AC and AM showed that only three common ingredients are palmitic acid(27.12% and 19.31%),octadecanoic acid(5.22% and 9.6%),and pentadecanoic acid(0.22% and 0.34%).Besides,linoleic acid(16.89%)and stigmastanol(1.68%)of AC is not detected in AM,stigmasterol(6.81%)and campesterol(1.34%)are not detected in AC.5、We identified 36 and 26 chemical compounds from LC-MS analysis of ethyl acetate extract part and n-butyl alcohol extract part.Its main constituents are organic acid and flavonoids together with many different ingredients in ethyl acetate extract part.There are exsited in four ingredients(triterpenoid saponins,flavonoids,alkanoids,and saccharides)containing pharmacological activity.6、According to molecular docking and ADMET forecastion,Triterpenoids & saponins and organic acids possess better pharmacodynamics activity due to high docking score,beter water solubility,well human intestinal sbsorption,non-hepatotoxicity,no cytochrome P4502D6 inhibition,good blood brain barrier permeability,and low protein biding ratio.But alkaloids has weak effect due to their hepatotoxicity,rejection capability of cytochrome P450 2D6,and worse blood-brain barrier permeability,this result should be paid attention in development and research.7、According to the result of anti-hepatic fibrosis in vitro,petroleum ether part of AC opssesses optimal inhibition effect to HSC-T6(P<0.001),the inhibiting rate was 87%.8、According to the results of anti-hepatic fibrosis in vivo,n-butyl alcohol part and petroleum ether part of AC has better curative effect in preventing and treating the formation of liver fibrosis,respectively.Comparing to model group,they can significantly reduce glutamic oxalacetic,glutamic pyruvic transaminase of serum.At the same time,they can obviously decrease hydroxyproline,monoamine oxidase,laminin,hyaluronic acid,Type Ⅲ procollagen,collage type-Ⅳ,and transforming growth factorβ1(P<0.05).9、According to the result of anti-hepatic fibrosis in vitro,linoleic acid opssesses optimal inhibition effect to HSC-T6(P<0.001),the inhibiting rate was 90%.Conclusion:1、According to resources investigation,we clear and definite that AM mainly distribute Guangxi and Guangdong.And its fake stamp and adulterant include Abrus mollis Hance,Abrus precatorius L.,Desmodium microphyllum(Thumb)DC.,Breynia vitis-idaed(Burm.f.)C.E.C.Fischer.2、According to morphological identification and microscopic identification of AC and AM,we explicit morphological and microscopy difference.This provides the basis for identifying correctly AC.3、According to component analysis of different parts,virtual screening of known chemical compounds,and experiments in vitro and in vivo,the result showed that n-butyl alcohol part and petroleum ether part of AC has better curative effect in preventing and treating the formation of liver fibrosis,respectively(P<0.05).What’s more,we verify that organic acids of AC have anti-liver fibrosis through in vitro experimental method... |
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