The Culture Supernatant Of Bone Marrow Mesenchymal Stem Cells On Treatment Of Liver Fibrosis Induced By Hepatic Stellate Cells

Objective Liver fibrosis is a repair reaction result that is due to a variety of acute or chronic inflammatory stimulation liver damage.Hepatic stellate cells(HSC)activation is considered to be a central part of liver fibrosis.The studies has been proved that bone marrow mesenchymal stem cells(BMSCs)transplantation therapy can improve or even reversal of liver fibrosis,but its mechanism is still unclear.This study through the culture supernatant of human bone marrow mesenchymal stem cells(HBMSCs)with HSC,to observe cell cycle of HSC and to detect the expression of matrix metalloproteinases-1(MMP-1),matrix metalloproteinases-2(MMP-2)and tissue inhibitors of metalloproteinase-1(TIMP-1).Methods Under the aseptic conditions,to extract liver cirrhosis volunteers'bone marrow 5ml,using density gradient centrifugation separation was used to collect and identify HBMSCs,training in 25cm2 the bottle of plastic,and HBMSCs culture supernatant was collected after 7d,added in HSC and then HSC were cultured 24h and 48h.Flow cytometry was to observe cell cycle of HSC,gelatin zmography method was to detect the expression of MMP-2 protein,ELISA was to detect the expression of MMP-1 and TIMP-1 protein,Real-time quantitative PCR method was to detect the expression of MMP-2,TIMP-1 mRNA.Results HSC in co-culture group has a significant increase in G1 phase cells,S phase cells were significantly reduced compared with HSC in alone group after 48h.Gelatin zmography method result are shown compared with HSC in alone group after 24h,48h,expression of MMP-2 in HSC of the co-culture group decreased significantly,differences were statistically significant(p<0.01).ELISA results are shown that expression of MMP-1 in HSC of the co-culture group increased significantly and expression of TIMP-1 in HSC of the co-culture group decreased significantly,compared with the HSC in alone group and differences were statistically significant(p<0.05).Real-time quantitative PCR result are shown compared with HSC in alone group after 24h,48h,expression of MMP-2 mRNA and TIMP-1 mRNA in HSC of the co-culture group decreased significantly,differences were statistically significant(p<0.01).Conclusion Liver cirrhosis volunteers' BMSCs has the effect of against hepatic fibrosis.According to the experimental results indicated that HBMSCs can stop HSC activation,play the roles to improve and treatment of liver fibrosis,also for further use patients with liver cirrhosis' BMSCs autologous transplantation in the treatment of liver cirrhosis provides an important theory basis.