选择性环氧合酶-2抑制剂对HSC-T6细胞增殖的抑制作用

目的 观察选择性环氧合酶-2抑制剂NS-398对肝星状细胞系HSC-T6细胞增殖的抑制作用,并探讨其可能的机制. 方法采用不同浓度的NS-398作用于HSC-T6,四甲基偶氮唑盐法检测HSC-T6细胞增殖情况,乳酸脱氢酶法检测NS-398对HSC-T6细胞的毒性作用、流式细胞仪检测HSC-T6细胞周期的改变,以EliVisionTMplus细胞免疫化学法检测HSC-T6中COX-2、增殖细胞核抗原(PCNA)蛋白表达的变化.多组数据间的比较采用单因素方差分析或独立样本T检验,计数资料采用χ2检验. 结果在20～160μamol/L浓度范围内,NS-398以剂量和时间依赖性地抑制HSC-T6增殖(P<0.01).90、120、150 μmol/L的NS-398作用HSC-T6 48 h后,细胞周期分析结果显示S期细胞稍增多,G2/M期细胞显著增加,各组之间细胞周期构成差异具有统计学意义(χ2=12.35,P<0.05).以120 μmol/L NS-398作用HSC-T6 48 h后,PCNA阳性细胞百分比为28.91%±0.11%,与对照组(85.99%±0.13%)比较,差异具有统计学意义(P<0.01);COX-2的阳性细胞百分比为13.80%±0.43%,与对照组(14.07%±0.59%)比较,差异无统计学意义(P>0.05).结论 NS-398可以显著抑制HSC-T6增殖,使其发生G2/M期阻滞,其作用具有剂量与时间依赖性,NS-398可能通过抑制HSC中PCNA的表达而抑制其增殖,但小影响COX-2的表达.

The role and mechanisms of cyclooxygenase-2 inhlbitors on the proliferation of hepatic stellate cell

Objective To observe the effects of NS-398 on proliferation of hepatic stellate cells (HSCs) in vitro, and to investigate the possible molecule mechanism. Methods HSCs were incubated with different concentrations of NS-398. The effects of NS-398 on cell proliferation was detected by MTT colormetric assay. The cell cycle of HSCs was analyzed by Flow Cytometry (FCM), cyclooxygenase-2 (COX-2) and proliferating cell nuclear antigen (PCNA) proteins in HSCs were detected by immunocytochemistry. Re-suits Administration of 20-160 μmol/L NS-398 significantly inhibited HSCs proliferation in dose-depen-dent manner compared with the control group (P<0.01). After treated with NS-398 at concentrations of 90, 120, and 150 μmol/L for 48 h, the number of HSCs in G2/M phase increased and the number of HSCs in G0/G1 phase decreased (P<0.05); Incubated with 120 μmol/L NS-398 for 48 h, percentage of masculine cell of PCNA was 28.91%±0.11%, which was significantly lower than that of the control group (85.99%±0.13%) (P<0.01). Percentage of masculine cell of COX-2 was 13.80%±0.43%, which was not signifi-cantly different from that of the control group (14.07%±0.59%) (P>0.05). Conclusions NS-398 could inhibit the proliferation of HSC-T6 and arrest HSCs in G2/M phase. Down-regulation of PCNA protein may partially accounted for the proliferation inhibition effect on HSCs induced by NS-398.