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目的 研究复方甘草酸苷(SNMC)保护人肝母细胞瘤HepG2细胞,减轻细胞凋亡的作用和机制。 方法 肿瘤坏死因子α(TNFα)和放线菌素D(ActD)诱导HepG2细胞凋亡后,分别加入0、2、2 0、100、200、800μg／ml终浓度的SNMC作用12 h。流式细胞仪分析凋亡细胞百分率;电镜观察细胞超微结构的变化;琼脂糖凝胶电泳观察细胞内片段化DNA的形成;Western blot检测此过程中细胞内凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)及B细胞淋巴瘤／白血病-2基因(Bcl-2)、B细胞淋巴瘤／白血病-2基因关联x蛋白(Bax)的表达情况。 结果 不同浓度SNMC处理TNF α等作用的HepG2细胞12 h后,随药物浓度增加,HepG2细胞凋亡率降低,凋亡特有的细胞内片段化DNA形成减少,细胞内Caspase-3相对分子量32×103的酶原表达增加,17×103活性成分表达减少,同时Bcl-2表达增加,Bax表达减少。另外,电镜下可见模型组细胞呈现凋亡后典型核浓缩,而100μg／ml药物保护组未见此现象。 结论 SNMC可抑制TNF α和ActD诱导的HepG2细胞凋亡,其机制可能通过调控凋亡相关蛋白的表达而实现。
Objective To study the effect and mechanism of compound glycyrrhizin (SNMC) on protection of hepatoblastoma HepG2 cells and apoptosis. Methods HepG2 cells were induced by tumor necrosis factor α (TNFα) and actinomycin D (ActD), and then treated with SNMC at final concentration of 0, 2, 2, 100, 200, 800 μg / ml for 12 h. The percentage of apoptotic cells was analyzed by flow cytometry. The changes of cell ultrastructure were observed by electron microscope. The formation of fragmented DNA was observed by agarose gel electrophoresis. The protein level of apoptosis related protein cysteine Caspase-3, Bcl-2, and Bax expression in B cell lymphoma / leukemia-2 gene. Results After HepG2 cells were treated with different concentrations of TNF-α for 12 h, the apoptotic rate of HepG2 cells decreased and the number of apoptotic-specific fragmented DNA decreased. The relative molecular mass of Caspase-3 was 32 × 103 Of the enzyme expression increased, 17 × 103 active ingredient expression decreased, while Bcl-2 expression increased, Bax decreased. In addition, under the electron microscope, the cells in the model group showed typical nuclear condensation after apoptosis, while the 100 μg / ml drug-protective group did not observe this phenomenon. Conclusion SNMC can inhibit the apoptosis of HepG2 cells induced by TNFα and ActD. The mechanism may be through regulating the expression of apoptosis related proteins.