二氢姜黄素体外处理BRL-3A细胞对细胞凋亡通路的影响

doi:10.3969/j.issn.1672-5069.2020.05.005

摘要/Abstract

摘要： 目的研究二氢姜黄素(DHC)对棕榈酸(PA)诱导的大鼠肝BRL-3A细胞线粒体凋亡通路的影响。方法建立PA诱导的非酒精性脂肪性肝炎(NASH)细胞模型,将细胞分为对照组(正常培养基)、模型组(0.25 mM PA处理)和DHC处理组(16、32和64μM DHC处理),检测细胞培养上清乳酸脱氢酶(LDH)含量,采用流式细胞仪检测细胞凋亡,检测细胞线粒体膜电位、凋亡相关蛋白酶Caspase-3和Caspase-9活性,采用Western blotting 法检测线粒体凋亡相关蛋白Bcl-2和Bax蛋白表达量。结果模型组细胞上清LDH、Caspase-3和Caspase-9活性分别为(100.1±4.2)%、(1.0±0.2)%和(1.0±0.1)%,均显著高于对照组,而线粒体膜电位为(0.5±0.1)%,显著低于对照组;32 μM DHC处理组LDH、Caspase-3和Caspase-9活性分别为(84.4±2.4)%、(0.8±0.1)%和(0.7±0.1)%,均显著低于模型组(P<0.05),而线粒体膜电位为[(0.6±0.1)%,显著高于模型组(P<0.05);与对照组比,模型组细胞凋亡率显著增加,而经DHC处理组,细胞凋亡率比模型组显著降低;模型组Bcl-2蛋白相对表达量为(1.0±0.1),显著低于对照组,而Bax蛋白相对表达量为(1.1±0.1),显著高于对照组;32 μM DHC处理组和64 μM DHC处理组Bcl-2蛋白相对表达量分别为(1.3±0.1)和(1.9±0.2),均显著高于模型组(P<0.05),而Bax蛋白相对表达量分别为(0.7±0.1)和(0.6±0.1),均显著低于模型组(P<0.05)。结论 DHC可能通过抑制线粒体介导的凋亡通路,缓解由PA诱导的BRL-3A细胞凋亡。

关键词: BRL-3A细胞, 二氢姜黄素, 非酒精性脂肪性肝炎, 线粒体, 凋亡通路 , ,

Abstract: Objective The aim of this study was to investigate the effects of dihyd rocurcumin (DHC) on apoptotic pathway in palmitic acid (PA)-induced BRL-3A cell apoptosis in vitro. Methods The non-alcoholic steatohepatitis (NASH) cell model was established by PA induction in BRL-3A cells. The survival rate of BRL-3A cells was detected by MTT assay. The cells were divided into control with normal medium, model (0.25 mM PA) and DHC intervention group with 16, 32, 64 μM DHC culture. The content of lactate dehydrogenase (LDH) in cell culture supernatant was detected. The apoptosis was detected by flow cytometry. The cell mitochondrial membrane potential (MMP), activity of apoptosis-related protease Caspase-3 and Caspase-9 were detected and the protein expression of mitochondrial apoptosis-related proteins Bcl-2 and Bax was detected by Western blotting. Results The supernatant LDH level, activities of Caspase-3 and Caspase-9 in model group were (100.1 ± 4.2)%, (1.0 ± 0.2)% and (1.0 ± 0.1)%, much higher than in the control, while the MMP was lower than that in control group ; the LDH, Caspase-3 and Caspase-9 in 32 μM DH-intervened cells were (84.4±2.4)%, (0.8±0.1)% and (0.7±0.1)%, significantly lower than those in the model (P<0.05), whilethe MMP was (0.6±0.1)%, much higher than that in the model (P<0.05); the apoptotic rate in the model group was significantly increased as compared to that in the control, while it decreased greatly as compared with model in DHC-intervened group; the relative expression of Bcl-2 protein in model group was lower than that in control group , while the relative expression of Bax protein was higher than that in control group , but the relative expression of Bcl-2 protein in 32 μM DHC- treated group and 64 μM DHC-treated group were (1.3 ± 0.1) and (1.9 ± 0.2), much higher than those in the model group (P<0.05), and the relative expression of Bax protein were (0.7 ± 0.1) and (0.6 ± 0.1), much lower than those in the model group (P<0.05). Conclusion DHC might relief the apoptosis caused by PA inBRL-3A cells by inhibiting the mitochondria-mediated apoptotic pathway.

Key words: BRL-3A cells, Dihydrocurcumin, Non-alcoholic steatohepatitis, Mitochondria, Apoptotic pathway

吴刚, 陈训军, 张万里, 李芬. 二氢姜黄素体外处理BRL-3A细胞对细胞凋亡通路的影响[J]. 实用肝脏病杂志, 2020, 23(5): 622-625.

Wu Gang, Chen Xunjun, Zhang Wanli, et al.. Effects of dihyd rocurcumin on apoptotic pathway in palmitic acid-induced BRL-3A cell apoptosis in vitro[J]. Journal of Practical Hepatology, 2020, 23(5): 622-625.

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