| Alcohol consumption in the world is increasing year by year.The number of deaths due to alcohol consumption accounts for 5.9% of the total deaths each year.Among them,acute alcoholism and alcoholic liver injury are the most harmful.Therefore,the development of anti-alcoholism and liver protection drugs is imminent.It is an important research direction to extract safe and effective antioxidant substances from natural plants to develop anti-alcoholism and liver-protection drugs.Clinacanthus nutans is a common medicinal and food plant in Southeast Asian countries.It has various physiological functions related to the principle of anti-alcoholism and liver protection,such as antioxidant,anti-oxidative stress,anti-inflammatory,etc.Flavonoids,polyphen ols and polysaccharides in Clinacanthus nutans leaves have the potential of anti-alcoholism and liver protection.Based on this,this paper first analyzed the nutrient components of the Clinacanthus nutans leaves.Then screened the anti-alcoholism functional components of Clinacanthus nutans leaves extract,and compared the antioxidant ability of the selected components.Then,the liver protection function of anti-alcoholism components was explored with the model of acute liver injury induced by alcohol and carbon tetrachloride.Finally,the components were separated by various chromatographic columns to obtain monomers,which provided theoretical basis for the rational development of the resources of Clinacanthus nutans leaves.The main contents and results of this paper are as follows:Firstly,the main nutrient components of the dried leaves of Clinacanthus nutans were determined.The results showed that the content of ash,protein,fat and total dietary fiber were 13.5%,25.3%,2.3% and 22.4% respectively.Total flavonoids 11.04 mg/g,polyphenols 12.62 mg/g,and polysaccharides 42.66 mg/g.It contained 7 essential amino acids and 10 non-essential amino acids.The values of EAA/NEAA and EAA/TAA were 0.62 and 0.38 respectively,which were equivalent to the ideal values recommended by FAO/WHO.At the same time,it contained 14 kinds of fatty acids,the linolenic acid accounted for 55%.In addition,it contained potassium(26898.1 mg/kg),calcium(21701.4 mg/kg),iron(574.6 mg/kg),zinc(36.9 mg/kg),selenium(1.33 mg/kg)and other trace elements.Secondly,the Clinacanthus nutans leaves was extracted with 75% ethanol to get the ethanol extract(P1),the residue was extract with the deionized water to get the aqueous extract(P2).The anti-alcoholism effect of P1 and P2 was tested by animal experiments.P2 performed better,and then separated by resin,three parts were obtained: A(water elution),B(10-50% alcohol elution)and C(70-90% alcohol elution).B showed the best effect of anti-alcoholism,the latency of drunkenness was significantly prolonged(P < 0.01),the sleeping time of drunkenness was significantly shortened(P < 0.05).The ADH and ALDH activities in liver of mice were significantly increased(P < 0.01 or P < 0.05),and the blood alcohol content of mice was significantly decreased by B(P < 0.01).The antioxidant activities of P1 and P2 were compared by ABTS,DPPH and FRAP antioxidant assays.The results showed that the antioxidant ability of P1 was stronger than that of P2,B and C was stronger than A.This indicated that the antioxidant activity of Clinacanthus nutans leaves was related to their antioxidant ability.Then,two acute liver injury models induced by alcohol and carbon tetrachloride were used to study the liver protection function of B.It was found that the hair color,stool and mental state of mouse pretreated with B were better than those of the model.The enlargement of liver and spleen was significantly inhibited by B(P < 0.01 or P < 0.05).B significantly reduced the activity of AST and ALT in serum(P < 0.05),and maintained the structure of hepatocyte membrane and mitochondria.Compared with the model group,the activities of SOD and GPX in the liver of group B were significantly increased(P < 0.05),the amount of MDA was significantly decreased(P < 0.05),the antioxidant capacity of the liver was improved,and the lipid peroxidation was effectively inhibited.It showed that B could protect the liver.Finally,B was separated by MCI column,and B1 and B2 were obtained.Then B1 and B2 were separated by ODS columns to get monomer compound.Three monomer compounds obtained from B: I,II,III.Two monomer compounds were obtained from B2: IV,V.The structure of the monomer compounds were analyzed by NMR.The result showed that compound I and IV was 3',4',7-trihydroxy-5-hydroxymethyl-6-C-beta-D-glucose dihydroflavonol glycosides,compound II was Indazole,compound III was Bartsioside,compound V was 2,3-dihydrosorbitol 3-oxo-beta-D-galactoside.I,II and V were first found in Clinacanthus nutans.In a word,Clinacanthus nutans leaves are rich in nutrient,which is a good source for supplementing essential amino acids and mineral elements such as calcium,iron,potassium,zinc and selenium.The Baltisides(iridoid terpenes)and dihydroflavones in Clinacanthus nutans leaves may be the functional ingredients of anti-alcoholism and liver protection,their effects and related mechanisms need further study. |
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