舒芬太尼对体外培养HepG 2 细胞增殖及细胞凋亡的影响

目的了解阿片类镇痛药舒芬太尼对体外培养肝癌细胞株HepG2细胞增殖、细胞周期及细胞凋亡的影响。方法体外培养人肝癌HepG2细胞,实验组在RPMI-1640培养液中加入不同浓度舒芬太尼（0.0001,0.001,0.01,0.1,1,10,20μmol·L-）1,对照组RPMI-1640培养液中不加舒芬太尼,分别孵育48小时后,采用MTT比色法检测HepG2细胞增殖活性,采用流式细胞技术检测舒芬太尼对HepG2细胞周期的影响,用Hoechst33258染色方法分析鉴定舒芬太尼对HepG2细胞凋亡的影响。结果舒芬太尼浓度≥1μmol·L-1时,细胞增殖活性较对照组显著下降（P〈0.05）;随着舒芬太尼浓度增加,G1期细胞比例逐渐增高,S期细胞比例逐渐降低。用Hoechst33258方法染色后在表面荧光显微镜下发现实验组部分HepG2细胞发生凋亡,当舒芬太尼浓度≥0.1μmol·L-1时,荧光显微镜下一个视野内凋亡细胞占总细胞数的比例较对照组显著增加（P〈0.05）。结论舒芬太尼在临床常用剂量范围内对肝癌细胞株HepG2细胞增殖和凋亡无明显影响,当浓度≥1μmol·L-1时可以抑制人肝癌细胞株HepG2细胞增殖,主要将细胞周期阻滞在G1期,当舒芬太尼浓度≥0.1μmol·L-1时可引起肝癌HepG2细胞凋亡。

The effect of sufentanil on the proliferation and apoptosis of human liver carcinoma cell line HepG2 in Vitro

Objective： To evaluate the effects of sufentanil on the proliferation, cell cycle and apoptosis of human liver carcinoma cell line HepG2 in vitro. Method Human liver carcinoma line HepG2 cells were cultured in vitro. The HepG2 cells of the test group were incubated in the RPMI-1640 medium with sufentanil at different concentration（0.0001, 0.001, 0.01, 0.1, 1, 10, 20μmol·L-1）, and the HepG2 cells of the control group were incubated in the RPMI-1640 medium for 48 hours. The level of the cell proliferation was evaluated with MTT method, and the cell cycle was detected with flow cytometry （FCM）, and the morphological changes of apoptosis cell was observed by fluorescence microscopy after staining by Hoechst33258. Results Compared with control group, the level of the cell proliferation in test group was apparently decreased（P﹤0.05）. With the concentration of sufentanil increasing, the ratio of G1 phase of HepG2 cells was significantly enhanced and the ratio of S phase of HepG2 cells was significantly decreased. Under the fluorescent microscopy after staining by the Hoechst33258, it showed part of HepG2 cells apoptosis in test group, and the rate of apoptotic cells was significantly increased when the concentration of sufentanil was over 0.1μmol·L-1 （P 0.05）. Conclusion The data suggested that sufentanil would not inhibit the level of the HepG2 cells proliferation and would not result in apoptosis when the concentration of sufentanil was in general range. Sufentanil could inhibit the level of the HepG2 cells proliferation when the concentration of sufentanil was over 1μmol·L-1, the inhibitory effect of sufentanil on HepG2 cells growth might be mediated by blocking cell cycle progression from G1 to S phase. Furthermore , sufentanil could result in human liver carcinoma HepG2 cells apoptosis when the concentration of sufentanil was over 0.1μmol·L-1.