| In China,ethnic minority people have developed their own national medical culture based on local natural resources.In the Tujia area of Enshi and Yichang,Hubei Province,three Crassulaceae plant Sedum lineare Thunb(SL),Sedum sarmentosum Bunge(SS),and Sedum emarginatum Migo(SE)were used to treat liver disease.The Tujia people drink hard liquor,and the liquor hurts the liver.However,it is not clear whether these three plants are effective for alcoholic liver disease(ALD).Objective:To investigate the effects of three common Crassulaceae plants,Sedum lineare Thunb,Sedum sarmentosum Bunge,and Sedum emarginatum Migo which are commonly used in Tujia nationality,against ALD in mice,and initially screen their active ingredients.Methods:1.The study on the effect of SL on acute alcoholic fatty liver disease in miceThe acute alcoholic fatty liver disease(AFLD)animal model was established by feeding Kunming female mice with alcohol once every 12 h,a total of 3 times.The mice in treatment groups were receiving fresh homogenate of fresh plants,which obtained by crushing stems and leaves.The mice were sacrificed after the final gavage for 4 hours.The serum of mice was used to study the biochemical indexes of liver function.The serum concentrations of Aspartate aminotransferase(AST),Alanine aminotransferase(ALT),Triglyceride(TG),and Alkaline phosphatase(ALP)were determined by biochemical analyzer.For histological assays,one portion of liver tissue was subjected to oil red O staining to observe the accumulation of fat droplets,the other portion of liver tissue was cracked into homogenate to detect triglyceride.2.Therapeutic effect of SS on acute alcoholic fatty liver and the screening of active parts / componentsThe effect of SS on acute AFLD was tested by the same method as SL.After it was proved to be effective,the dried whole SS powder was heated under reflux to extract by solvents: petroleum ether,ethyl acetate,methanol and water,and the different polar extracting solution were heated and concentrated to obtain extracts.The aforementioned animal models were used to test and screen the extracts of various parts of SS and the anti-alcoholic fatty liver disease effect of sarmentosin,a known hepatoprotective component.3.Therapeutic effect of SE on early alcoholic hepatitis and screening of active partsThe same method was used as SL to test the effect of SE on acute AFLD.After it was proved effective,the dried SE powder was extracted under heating and reflux by different polar solvents: petroleum ether,ethyl acetate,methanol and water.The extracting solution were heat and concentrated to obtain an extract.It was found that the content of polysaccharides in the water extract was relatively high,and we used polysaccharides instead of water extracts for experiment.Considering that polysaccharides generally have a good immunomodulatory effect,we tested the therapeutic effect of each part of SE in a mouse model of early alcoholic hepatitis(AH).Mice were given 5 g/kg of alcohol every 12 hours,a total of 9times,and lipopolysaccharide LPS was injected intraperitoneally at the 3rd and 7th times.After 4 hours of the last gavage,the mice were sacrificed.The serum was used to detect the liver function of biochemical indicators,and the changes of liver function indicators in the serum were analyzed.The liver tissue was used for H & E staining to observe the distribution and damage of lipid droplets in the liver tissue.A part of the liver tissue homogenate was used to detect the triglyceride content,which was used to prove the lipid accumulation in the liver tissue.A small part of liver tissue was taken for transmission electron microscopy to observe the changes of the internal structure and shape of liver cells.Result:1.The SL could reduce the lipid accumulation in liver tissue caused by alcohol,but could not reduce the serum triglycerides level.Serum biochemical indicators showed that compared with the blank control group,the model group had significantly higher serum triglyceride levels(p <0.05).Low doses of SL could improve the elevated serum triglyceride levels,but high doses had no therapeutic effect.Tissue oil red O staining results showed that after treatment,the distribution of lipid droplets in liver tissue was significantly reduced.2.The SS could effectively inhibit the increase of serum triglyceride level and liver lipid accumulation caused by alcohol,and its active ingredient was a large polar part.SS homogenate can effectively reduce alcohol-induced hyperlipidemia.Liver tissue homogenization results showed that high dose of SS was more effective than low dose in reducing triglyceride accumulation in liver tissue.Oil red O staining of liver tissue further confirmed this result.The extracts of small polar solvents petroleum ether and ethyl acetate have no therapeutic effect on the rise of triglycerides in serum and liver tissue caused by alcohol,while methanol extract and aqueous extract can effectively reverse the rise in serum triglycerides caused by alcohol,and aqueous extract had a better inhibitory effect on the accumulation of triglycerides in liver tissue,and liver function indicators also showed that it played a protective role in the liver.H & E staining of tissue sections also reflected its effectiveness against alcohol-induced liver tissue damage and inflammatory foci.3.The SE could effectively fight the increase of serum triglyceride level caused by alcohol and the accumulation of lipid in liver tissue and the production of inflammatory foci.Its effective part against early alcoholic hepatitis was polysaccharideThe high dose of SE had a certain reversal effect on the increase of serum triglycerides caused by alcohol,and it had a better effect on the accumulation of lipid droplets in liver tissues.Oil red O staining proved this result.In all parts,the crude polysaccharide did not significantly reverse the increase of serum triglyceride levels in early AH,but it could significantly alleviate the increase of inflammatory foci in liver tissue.From the above experimental results,the effects of the three plants against ALD were not exactly the same.For the effect of treating alcohol-induced hyperlipidemia,SS was superior to SE,while SL had no therapeutic effect on the increase of serum triglycerides levels.For the treatment of lipid accumulation in liver tissue caused by alcohol,SE was superior to SS better than SL,and sarmentosin was not the active ingredient of SS against ALD.Therefore,further separation of the components of SS and SE was performed,and the parts with better therapeutic effects of both were aqueous extract,and the content of polysaccharides in the aqueous extract was found to be higher by testing.Conclusion:This experiment focused on three plants of Crassulaceae,SL,SS and SE to treat ALD.SL was less effective in treating AFLD,and SS and SE played an effective role in reducing the levels of triglycerides serum and liver.Compared with other extracts,the aqueous extract had the best therapeutic effect on reversal liver damage caused by alcohol.The polysaccharides of SE had effect against early AH,but the material basis of Crassulaceae anti-alcoholic fatty liver disease was not yet clear.This experiment had laid a certain foundation for further exploring the material basis and mechanism of Crassulaceae plants against ALD. |
