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目的:观察选择性环氧合酶-2抑制剂NS-398对肝星状细胞(HSC)凋亡及凋亡相关基因的影响。方法:采用不同浓度的NS-398作用于HSC-T6,应用流式细胞术和透射电镜检测HSC凋亡,细胞免疫化学法检测凋亡HSC中凋亡相关基因p53、bcl-2蛋白表达的变化。结果:90、120、150μmol/LNS-398作用HSC-T6 48h后,流式细胞术检测到HSC凋亡,各组凋亡指数分别为(10.5±0.015)%、(13.2±0.010)%和(17.3±0.012)%,与对照组(4.3±0.006)%比较,差异有统计学意义(P<0.01);透射电镜观察到细胞皱缩、核染色质浓缩沿核膜排列等细胞凋亡特征性改变;以120μmol/L NS-398作用HSC-T6 48h后,凋亡相关蛋白p53、bcl-2的阳性细胞百分率分别为(82.06±0.14)%、(34.20±0.77)%,与对照组(14.06±0.24)%、(96.66±0.79)%比较,差异有统计学意义(P<0.01)。结论:NS-398可以剂量依赖性诱导HSC凋亡;上调凋亡相关基因p53的表达,下调bcl-2的表达可能是NS-398诱导HSC凋亡的机制之一。
Objective: To observe the effect of NS-398, a selective cyclooxygenase-2 inhibitor, on apoptosis and apoptosis-related genes in hepatic stellate cells (HSC). Methods: The HSC-T6 cells were treated with NS-398 in different concentrations. The apoptosis of HSC-T6 cells was detected by flow cytometry and transmission electron microscopy. The expressions of p53 and bcl-2 protein were detected by immunocytochemistry . Results: HSC-T6 treated with 90, 120 and 150μmol / L NS-398 for 48h, the apoptosis rate of HSC was detected by flow cytometry, the apoptotic indexes were (10.5 ± 0.015)%, (13.2 ± 0.010)% and 17.3 ± 0.012)%, compared with the control group (4.3 ± 0.006)%, the difference was statistically significant (P <0.01); the characteristic of cell apoptosis such as cell shrinkage and nuclear chromatin condensation along the nuclear membrane was observed by transmission electron microscope (82.06 ± 0.14)% and (34.20 ± 0.77)%, respectively. The percentage of positive cells of apoptosis-related protein p53 and bcl-2 in 120μmol / L NS-398 treated HSC- ± 0.24)%, (96.66 ± 0.79)%, the difference was statistically significant (P <0.01). Conclusion: NS-398 can induce HSC apoptosis in a dose-dependent manner. Upregulation of p53 gene expression and down-regulation of bcl-2 may be one of the mechanisms of HSC apoptosis induced by NS-398.