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目的研究自噬在槲皮素(quercetin,Que)诱导的人肝癌细胞系SMMC-7721死亡中的作用。方法采用四甲基偶氮唑蓝法测定槲皮素对SMMC-7721细胞的抑制作用;单丹磺酰戊二胺、免疫荧光法观察给药后自噬的发生;乳酸脱氢酶(LDH)漏出率、蛋白印迹法观察槲皮素诱导的SMMC-7721细胞死亡的分子机制。结果槲皮素对SMMC-7721生长有显著抑制作用,呈明显的时间、剂量依赖性;槲皮素可激活SMMC-7721细胞的LC3的表达、诱导自噬的发生;乳酸脱氢酶漏出率实验结果表明,在槲皮素给药前用3-甲基腺嘌呤(3-MA)阻断自噬或者氯喹碱化溶酶体,可增强槲皮素对SMMC-7721细胞的细胞毒作用;蛋白印迹结果显示,槲皮素可使SMMC-7721细胞溶酶体组织蛋白酶B的表达明显增强。结论槲皮素能明显抑制SMMC-7721细胞的生长,并诱导其发生自噬;自噬激活能降低槲皮素对SMMC-7721细胞的毒性;自噬的发生与溶酶体酶组织蛋白酶B表达增强相关。
Objective To investigate the role of autophagy in the quercetin (Que) -induced death of human hepatoma cell line SMMC-7721. Methods The inhibitory effect of quercetin on SMMC-7721 cells was determined by MTT assay. The monosandamyl pentamethylene diamine was used to detect the occurrence of autophagy after administration. The levels of lactate dehydrogenase (LDH) Leakage rate and Western blotting were used to observe the molecular mechanism of quercetin-induced SMMC-7721 cell death. Results Quercetin significantly inhibited the growth of SMMC-7721 cells in a time-and dose-dependent manner. Quercetin activated the expression of LC3 in SMMC-7721 cells and induced the release of autophagy. Lactate dehydrogenase leakage rate test The results showed that 3-methyladenine (3-MA) blocking autophagy or chloroquine alkaline lysate before quercetin administration increased the cytotoxic effect of quercetin on SMMC-7721 cells; protein Western blotting results showed that quercetin significantly enhanced the expression of lysosomal cathepsin B in SMMC-7721 cells. Conclusion Quercetin can significantly inhibit the growth of SMMC-7721 cells and induce autophagy. Autophagy activation can reduce the toxicity of quercetin to SMMC-7721 cells. The occurrence of autophagy and the expression of lysosomal enzyme cathepsin B Enhance related.