The Effect Of Sini Ruangan Recipe On The Four Items Of Liver Fiber, MMP-1 And TIMP-1 Of Human Hepatic Stellate Cells Under The Action Of Acetaldehyde

Objective:Observation the effects of Sini Ruangan recipe on Liver fiber four(CG,HA,LN,PC?)and MMP-1,TIMP-1 in human HSC under the action of acetaldehyde.To investigate the mechanism and effect of anti liver fibrosis.Methods:1.LX-2 cells(1x105/ml)were cultured into a sterile 96-hole culture plate,200ul/hole,and cultured in incubator for 24 hours,until the cells covered with monolayers,pour out the culture medium.Different concentrations of acetaldehyde were added to the normal medium for 24 hours,48 hours,72 hours(change the liquid once per 24 hours),the OD value of cells was measured by CCK-8 method,and the degree of damage of LX-2 cells by acetaldehyde was analyzed.The concentration and time of acetaldehyde with the highest cell survival rate were selected as the injury concentration and time.2.LX-2 cells were inoculated into the sterile 96-well plate,and the cells covered with monolayers.The LX-2 cells randomly divided into Normal group(culture of normal culture medium),Acetaldehyde group(cell culture medium containing250mmol/L acetaldehyde),cultured 72 hours(change the liquid once per 24hours).Two groups of cell culture liquid supernatant was collected to be measured.Then,the LX-2 cells in Acetaldehyde group were divided into Tuoli group,Sini group,Qingre group and Fuzheng group;Sini group,Qingre group,Fuzheng group make use of the corresponding drug-containing serum culture,Tuoli group and Normal group make use of the normal medium culture,cultured for 24 hours,the cell culture liquid supernatant was collected to be measured.3.Used ELISA kit to detecting the liver fiber and MMP-1 TIMP-1 in the supernatant of cell culture medium.Results:1.Different concentrations and time of acetaldehyde damage to LX-2 cells was difference,least damage,the cell survival rate was the highest in 250mmol/L acetaldehyde culture for 72 hours.2.Observation of 200 times under inverted microscope in laboratory,compared with Normal group,the density of cells in Acetaldehyde group was obviously sparse,cell morphology remains unchanged.3.Compared with Normal group,Acetaldehyde group on PC?was increased significantly(P?0.01);had no difference in the content of CG,HA,LN,MMP-1 and TIMP-1(P>0.05).4.Compared with Tuoli group,the levels of HA,PC? and TIMP-1 decreased significantly(P?0.01,P?0.05),CG,LN and MMP-1 were no statistical difference(P>0.05)in Sini group.5.Sini group,Qingre group,Fuzheng group had no difference among the three groups(P>0.05).Conclusion:Sini Ruangan recipe can reduce the expression of indicators on HA,PC? and TIMP-1 in damaged LX-2 cells,which may play a role in antagonizing liver fibrosis.