对叶百部总生物碱保护H_(2)O_(2)诱导V79细胞损伤的作用及机制研究  

Study on the protective effects and mechanism of total alkaloids from Stemona Tuberosa Lour by H_(2)O_(2)-induced injury in V79 cells

作　　者：韦卫宁 许立拔 王孝勋 杨雅钧 何石燕 周彩燕 WEI Wei-ning;XU Li-ba;WANG Xiao-xun;YANG Ya-jun;HE Shi-yan;ZHOU Cai-yan(College of Pharmacy, Guangxi University of Chinese Medicine, Nanning 530200, China;Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine, Nanning 530200,China)

机构地区：[1]广西中医药大学药学院,广西南宁530200 [2]广西壮瑶药重点实验室,广西南宁530200

出　　处：《陕西科技大学学报》2022年第4期74-80,共7页Journal of Shaanxi University of Science & Technology

基　　金：国家自然科学基金项目(81360640);广西南宁市科学研究与技术开发计划项目(20193120);广西壮瑶药协同创新中心科研项目(桂教科研〔2013〕20号);广西壮瑶药重点实验室科研基金项目(桂科基字[2014]32号);广西中医药大学研究生教育创新计划项目(YCXJ2021013)。

摘　　要：采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基溴化四唑(MTT)法测定对叶百部总生物碱(0、10、22、50、112和250μg/mL)和过氧化氢(0、0.5、1.0、2.0、3.0、4.0、5.0、6.0、7.0和8.0 mmol/L)对V79细胞的细胞毒性,并使用细胞活力测定、Hoechst 33258染色法、流式细胞术和Western blot法等方法观察对叶百部总生物碱对H_(2)O_(2)(5.0 mmol/L,24 h)所致V79细胞损伤的保护作用.结果显示,对叶百部总生物碱对V79细胞无明显细胞毒性,而H_(2)O_(2)抑制细胞增殖作用明显,呈剂量相关性,其IC_(50)值为4.42±0.45 mmol/L,而对叶百部总生物碱能显著降低H_(2)O_(2)(5.0 mmol/L,24 h)诱导的V79细胞毒性(P<0.05,P<0.01),可显著拮抗H_(2)O_(2)所致V79细胞凋亡作用(P<0.05,P<0.01),并能显著上调V79细胞中的Bcl-2蛋白表达,下调Bax和激活型半胱氨酸蛋白酶-3(Cleaved Caspase-3)蛋白表达(P<0.05,P<0.01).故认为对叶百部总生物碱对H_(2)O_(2)所致V79细胞凋亡具有良好的保护作用,其机制可能与下调Bax、Cleaved Caspase-3的表达和上调Bcl-2表达有关.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to measure the cytotoxicity of total alkaloids from Stemona Tuberosa Lour(0,10,22,50,112 and 250μg/mL)and H_(2)O_(2)(0,0.5,1.0,2.0,3.0,4.0,5.0,6.0,7.0 and 8.0 mmol/L),and cell viability assay,Hoechst 33258 staining,flow cytometry and Western blot assay were used to evaluated the protective effects of the total alkaloids against H_(2)O_(2)(5.0 mmol/L for 24 h)to V79 cells.The results shows that total alkaloids from Stemona Tuberosa Lour had no cytotoxicity to V79 cells,whereas H_(2)O_(2) had dose-dependent cytotoxicity in inhibiting the proliferation of V79 cells,the IC_(50) value was 4.42±0.45 mmol/L;as well as the total alkaloids significantly decreased cytotoxicity to V79 cells induced by H_(2)O_(2)(P<0.05,P<0.01),inhibiting apoptosis of V79 cells induced by H_(2)O_(2)(P<0.05,P<0.01),up-regulating the expression of protein Bcl-2,and down-regulating the expression of Bax and Cleaved Caspase-3 in V79 cells(P<0.05,P<0.01).All in all,Total alkaloids of Stemona Tuberosa Lour have good protective effects against apoptosis in V79 cells induced by H_(2)O_(2),probably through downregulation of Bax and Cleaved Caspase-3 levels and upregulation of Bcl-2 level.

关 键 词：总生物碱 对叶百部 凋亡 V79细胞 H_(2)O_(2) 

分 类 号：R285[医药卫生—中药学]