Bone Marrow Mesenchymal Stem Cells Modulate The Expression Of Death Receptor-5in Hepatic Stellate Cells

Objective:To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on the protein expression of death receptor-5and Caspase-3in hepatic stellate cells(HSCs)by co-cultured,and explore the possible mechanisms involved.Methods:BMSCs were isolated from bone marrow in rats and grown,propagated in culture flask in the passages3.Primary cultured HSCs and fibroblast cells were recoveried and activated morphologically.The co-culture of BMSCs with HSCs was performed by using a6-well Transwell membranes.HSCs were seeded in the lower chamber with BMSCs and fibroblast cells(2×105cells/mL)were seeded onto the Transwell membrane of the inner chamber.Four groups were divided randomly:①SCs control group②Fibroblast control group③BMSCs co-culture group④Experimental group.Cell proliferation and apoptosis were determined by MTT assay and flow cytometry,respectively.Cell supernatants were harvested to determine the concentration of tumor necrosis factor-related apoptosis-inducing ligand by enzyme-linked immunosorbent assay(ELISA).The protein expressions of Caspase-3and death receptor-5in HSCs were determined by Western blot.The mRNA expression of death receptor-5was evaluated with RT-qPCR. Results:1.The inhibitory rates of HSCs proliferation with MSCs culture were significant higher than that of control groups and Experimental group at times24h,48h and72h(all P<0.01).2.After24h,48h and72h of co-culture,the apoptosis rates in BMSCs co-culture group were10.17%、15.15%and21.18%.The increased apoptosis rates of HSC in the MSCs group were significantly higher than those in the other three groups(all P<0.01).3.The concentrations of tumor necrosis factor-related apoptosis-inducing ligand in co-culture supernatants in each group at48h were the highest.And the concentrations of tumor necrosis factor-related apoptosis-inducing ligand in MSCs co-culture group were lower than the MSCs control group.4.The protein expressions of caspase-3in HSCs in BMSC co-culture group were higher than the other groups(all P<0.01).And the protein expressions of caspase-3in HSCs in BMSC co-culture group at72h were higher than that of24h or48h.5.The protein death receptor-5in HSCs in BMSC co-culture group were higher than the other groups(all P<0.01).And the protein expressions of death receptor-5in HSCs in BMSC co-culture group at72h were higher than that of24h or48h.6.The mRNA expression of death receptor-5in BMSCs co-culture group was higher than the other groups(P<0.01).And there was no significant difference between the HSCs control group and the fibroblast control group(P>0.05).Conclusions:BMSCs promote apoptosis,inhibit proliferation and increase the protein expressions of caspase-3and death receptor-5in rats activated HSCs perhaps in an tumor necrosis factor-related apoptosis-inducing ligand paracrine-dependent manner.