Effect of interfering RNA targeting plasminogen activator inhibitor-1 on the biological characters of hepatic stellate cells / 中华消化杂志

ABSTRACT

Objective To investigate the effect of synthetic small interfering RNA (siRNA) targe- ting plasminogen activator inhibitor-1 (PAI-1) on the biological characters of hepatic stellate cells (HSCs).Methods Synthetic siRNA targeting PAI-1 was transfected into HSC-T6 by lipofectamine package.Negative siRNA transfection and no transfection were used as negative control and blank con- trol respectively.After incubation with siRNA,total RNA and protein of HSC-T6 cells were extracted. The expression of PAI-1 gene was detected by reverse transcription-polymerase chain reaction and immuno- cytofluorescent.The proliferation of HSC-T6 was determined using MTT assay.HSC-T6 cycle and apoptosis were measured by flow cytometry.Content of typeⅠandⅢcollagen in the supernatants were determined by enzyme linked immunosorbent assay.Results The expressions of PAI-1 mRNA and pro- tein were markedly down-regulated in siRNA-transfected HSC-T6.The proliferation of HSC-T6 was inhibited by transfection of siRNA.The level of typeⅠcollagen in the supernatants decreased by (20.71?8.40)% and (37.97?6.40)%,repectively(F=42.69,P=0.0001) compared to those in the negative control at 48 and 72 hours.The level of typeⅢcollagen decreased by (35.98?4.60)% (F=105.52,P= 0.0001) at 72 hours.Conclusion Inhibition of PA-1 by siRNA may have a potential effect in prevention and treatment of hepatic fibrosis by inhibiting proliferation,promoting apoptosis of stellate cells.Therefore,it decreases the synthesis and the secretions of typeⅠandⅢcollagen.