The Role Of Nimesulide On The Rassf1Gene Expression In Rat Hepatic Stellate Cells

ObjectiveTo explore the effect of nimesulide on the expression of rassf1in TSC-6cells.Methods1.The logarithmic growth phase HSC-T6cell were cultured in RPMI-1640medium containing10%calf serum,37℃,5%CO2incubator. For6-8hours, adding200μmol/L、400μmol/L nimesulide and the RPMI-1640medium respectively(eachgroup with parallel holes).2.COX-2(cyclooxygenase-2) were evaluated by Immunocytochemistry methodat48h.3.TSC-6cell growth was assayed by MTT method at24h,48h,72h;4.Apoptosis were detected by flow cytometry at48h;5.Rassf1expression were evaluated by Real-time fluorescent quantitative PCR at48h;Results1. Immunocytochemistry results showed cyclooxygenase2expression positiverate were70.13±2.40％,40.13±4.40％and30.36±5.81%respectively in control,200μmol/L and400μmol/L nimesulide treated cells at48h (P <0.01);2.MTT results showed that400μmol/L nimesulide inhibited the proliferation ofTSC-6cells at48h;3.Flow cytometry showed the apoptotic rate in control,200μmol/L and400μmol/L nimesulide treated cells were10.23±1.34,25.23±7.7,65.87±2.89respectively at48h(P <0.01);4.Real-time fluorescent quantitative PCR results showed the expression of rassf1in HSC-T6cells was increased by nimesulide (200，400μmol/L) treatment (P <0.01)at48h.Conclusions1. Nimesulide may inhibit growth and induce apoptosis as well as increase rassf1 expression of TSC-T6;2.The effect of antiproliferation and induce apoptosis of Nimesulide may beassociated with its increasing rassf1expression in HSC-T6.