The Effect Of Hepatocyte Growth Factor On Proliferation And Apoptosis Of Hepatic Stellate Cells

Objective To evaluate the proliferation and apoptosis of rat hepatic stellate cells (HSCs) via hepatocyte growth factor (HGF) activated by hepatocyte growth factor activator (HGFA) in the co-culture system of Bone marrow mesenchymal stem cells (BMSCs) and HSCsMethods Cells were divided into the following groups:①HSCs control group:HSCs co-cultured with fibroblast cells.②HSCs blank group:HSCs cultured alone.③BMSCs blank group:BMSCs cultured alone.④Experimental group:HSCs+BMSCs.⑤HGFA intervention group:HSCs was intervented by70ng/ml of HGFA. Using transwell insert establish co-culture system in the plastic plate, The cultured period was maintained and cells were performed at24h,48h and72h. At the different time point, dynamic cells morphous were observed under the inverted phase contrast microscope. the surface markers of BMSCs and the apoptosis rate of HSCs were detected by flow cytometry. The expression of a-SMA of HSCs was immunohistochemically evaluated. The actived HGF (HGF-α chain) was evaluated by immunofluorescence staining. The proliferation of HSCs was tested by MTT assay. The concentration of HGF and HGFA were detected by enzyme-linked immunosorbent assay(ELISA).Results①MTT method showed no inhibitory action on HSCs proliferation rate stimulated by HGF alone compared to the HSCs blank group (p>0.05). while stimulated by HGFA with the concentration of70ng/ml for24h, The proliferation rate of HSCs was found significantly inhibited compared with that of the HSCs blank group (0.26±0.00VS0.13±0.04, P<0.01), as shown by does-dependent reduction.②The expression of HGF-α in experimental group (24h,48h:30.10±3.74,34.30±1.24) and HGFA intervention group (24h,48h:32.74±1.72,36.47±2.07) showed no difference at24h and48h (P>0.05), while lower in experimental group than in HGFA intervention group at72h (37.24±1.03VS40.44±0.77, P<0.05), and these groups were higher than all the control group at any time (P<0.05).③The apoptosis rate of HSCs in experimental group increased in24h,48h and72h, compared with HGFA intervention group, and this increase reached to its peak in72h (40.77%±1.16%VS33.35%±2.04%, P<0.01), this increased rate was higher in the above two group than in the control group at different time point (P<0.01).④The concentration of HGF in both experimental group and HGFA intervention group was time-dependent reduction, and lower than HSCs control group (P<0.05).⑤The concentration of HGFA in experimental group was higher than blank group at different time point(P<0.05).Conclusion The co-culture system of BMSCs and HSCs can inhibit the proliferation of HSCs and induce its apoptosis by promoting the secretion of HGFA and activating HGF.