Influence Of Human Umbilical Cord Mesenchymal Stem Cells On Hepatic Stellate Cells Proliferation

Objective To explore the mechanism of human umbilical cord mesenchymal stem cells (UCMSCs) to regulate the proliferation of hepatic stellate cells (HSCs-LX2) under co-culture in vitro.Methods Human UCMSCs and HSCs in the experimental group were cultured in the plastic culture plate(6holes) to establish the upper and lower double-cell co-culture system. HSCs were cultured alone as negative control group. Cell proliferation and apoptosis were determined by MTT assay and flow cytometry, respectively. Cell supernatants were harvested to determine the concentration of transforming growth factor betal (TGF-β1) by enzyme-linked immunosorbent assay (ELISA). The TGF-β1, Smad3and Smad7mRNA expression in HSC was determined by reverse transcription-polymerase chain reaction (RT-PCR) and the level of TGF-β1, Smad3and Smad7protein by Western blot..Results HSCs co-cultured with UCMSCs significantly inhibited HSC proliferation compared with the negative control group at24,48hours(P<0.05); Flow cytometry showed that HSCs apoptosis of co-culture group was increased compared with negative control group, and48h is more obvious (P<0.05). The concentrations of TGF-β1in co-culture supernatants in the experimental group was significantly lower than those in the supernatants of HSCs cultured alone, and48h is more obvious (P<0.05). After HSCs co-cultured with BMSCs for48hours, the expression of TGF-β1, Smad3mRNA and protein was reduced and the expression of Smad7mRNA and protein was increased compared with negative control group (P<0.05).Conclusion UCMSCs inhibited the proliferation of HSCs, possibly through inhibiting the TGF-β1and Smad3expression, increasing Smad7protein expression.