The Proliferation And Activation Of Aspirin On Cultured Hepatic Stellate Cells Line T6and Related Molecular Mechanisms

Objective To explore the impact and related mechanisms of the proliferation and activation of HSC through different concentration by design in vitro aspirin on cultured hepatic stellate cells line T6,to provide further theoretical basis for aspirin’s clinical application.Methods HSC-T6were cultured in DMEM medium which contain10%fetal bovine serum in37℃、5%CO2incubator. The impact of aspirin on cell proliferation was detected in different concentration (1,5,10,15mmol/L)with MTT.Morphological change of apoptosis cells were observed by light microscope.In accordance with the test results, the experiment was divided into the blank control group,aspirin group (1,3,5mmol/L), HSC-T6cells were intervened in vitro for24h,48h,72h, and a-SMA, Col I, COX-2,NF-κ B,PPAR-γ expressions were investigated with RT-PCR method.HSC was intervened in vitro for48h by1μmol/L LPS group、1μmol/L LPS combined different concentration ASA(1,3,5mmol/L)COX-2,NF-KB,PPAR-y expressions were investigated with RT-PCR method.Results1. Effect of aspirin on proliferation of HSC-T6cellsThe inhibitory rate of aspirin on HSC-T6cell proliferation increased the inhibitory rate of cell proliferation was not significantly (P>0.05).After48hour and72hour, aspirin can significantly inhibit HSC-T6proliferation, there are time-and concentration-dependent manner.2. Effect of aspirin on the mRNA expression of α-SMA、 Col I、 COX-2、 NF-κB、 PPAR-γ genes.(1) The expressions of α-SMA、 Col I、 COX-2and NF-κ B mRNA were significantly decreased compared with the control group after aspirin groups on HSC-T6for24h,48h,72h (P<0.05), and the reduction was significantly with enhanced concentration;And significantly decreased with the increase of the concentration between the two groups (P＜0.05).(2) Compared with the control group,the expression of PPAR-γ genes was significantly higher that showed significant differences among aspirin groups for24h,48h(P<0.05),but there was no significant differences between the two groups (P<0.05);Compared with the control group, there was no significant differences1mmol/L aspirin group for72h (P<0.05);Compared with the aspirin group (3,5mmol/L), there was a significant differences1mmol/L aspirin group for72h (P<0.05).(3) The expression of α-SMA/PPAR γ mRNA was significantly decreased compared with the control group after aspirin groups on HSC-T6for24h,48h,72h (P＜0.05), and the reduction was significantly with enhanced concentration, there was a significant difference between the two groups (P＜0.05)3. Effect of aspirin combined with LPS on the mRNA expression of COX-2、NF-κB、 PPAR-γ genes.(1) Compared with the control group,COX-2and NF-κ B mRNA expressions were significantly increased in LPS group (P <0.05);Compared with LPS group and the control group,the degree of inhibition of COX-2and NF-κB mRNA expressions were higher with ASA concentrations increased in LPS combined with ASA group(P <0.05).(2) Compared with the control group,PPAR-γ mRNA expression was significantly increased (P<0.05) in LPS group;Compared with the LPS group and the control group,the expression level of PPAR-gamma mRNA was stronger with increasing ASA concentration in LPS combined with ASA group.Conclusions1. Aspirin can inhibit the proliferation of HSC-T6cells.2. Aspirin can affect the activation of HSC-T6cells may be related to reduce the synthesis of liver fibrosis α-SMA and Col Ⅰ secretion.3. The study showed that the molecular mechanism of proliferation of aspirin on HSC-T6cells may be related to down-regulation NF-κ B mRNA and up-regulation of PPAR-γ mRNA.