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目的:探讨山楂提取物体外对肝癌细胞株HepG2凋亡的影响。方法:山楂乙醇提取,并采用不同质量浓度(0.1,0.2,0.4,0.8 g·L-1)山楂提取物处理HepG2细胞24,48,72 h。四甲基偶氮唑盐比色(MTT)法检测提取物对肝癌细胞活力的影响;流式细胞术检测提取物处理细胞48 h后的凋亡率;实时定量荧光PCR(real-time PCR)检测半胱氨酸蛋白酶-3(Caspase-3),B淋巴细胞瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax)基因表达;免疫印迹法(Western blot)检测激活型Caspase-3(CleavedCaspase-3),Bcl-2,Bax蛋白表达。结果:山楂提取物对HepG2细胞的活力有显著抑制作用(P<0.05),并以浓度及时间依赖的形式诱导细胞凋亡。山楂提取物可以显著促进Bax和Cleaved-Caspase-3基因和蛋白表达,抑制Bcl-2基因和蛋白表达。结论:山楂提取物抑制肝癌细胞HepG2的增殖,促进其发生凋亡,与Bax和Cleaved-Caspase-3的表达增加,Bcl-2/Bax降低有关。
Objective: To investigate the effect of hawthorn extract on the apoptosis of HepG2 hepatocellular carcinoma cell line in vitro. Methods: The hawthorn ethanol was extracted and the HepG2 cells were treated with hawthorn extracts of different concentrations (0.1, 0.2, 0.4 and 0.8 g · L -1) for 24, 48 and 72 h. The MTT assay was used to detect the effect of the extract on the viability of hepatocellular carcinoma cells. Flow cytometry was used to detect the apoptosis rate of the cells treated with extract for 48 h. The real-time quantitative PCR (real-time PCR) The expressions of caspase-3, Bcl-2 and Bcl-2 related protein B (Bax) were detected by Western blot. 3 (Cleaved Caspase-3), Bcl-2, Bax protein expression. Results: Hawthorn extract significantly inhibited the viability of HepG2 cells (P <0.05), and induced apoptosis in a time-and concentration-dependent manner. Hawthorn extract can significantly promote Bax and Cleaved-Caspase-3 gene and protein expression, inhibit Bcl-2 gene and protein expression. CONCLUSION: Hawthorn extract can inhibit the proliferation of HepG2 hepatocarcinoma cells and induce its apoptosis, which is related to the increase of Bax and Cleaved-Caspase-3 expression and the decrease of Bcl-2 / Bax.