紫草素对HepG2细胞增殖、凋亡和PI3K/Akt/NF-κB信号通路蛋白表达的影响*

doi:10.3969/j.issn.1672-5069.2020.04.005

摘要/Abstract

摘要： 目的研究紫草素对肝癌HepG2细胞增殖、凋亡和磷脂酰肌醇3-激酶(PI3K)/蛋白激酶(Akt)/核转录因子-κB(NF-κB)信号通路的影响。方法以0(对照)、1、2.5和5 μmol/L紫草素分别处理对数生长期的HepG2细胞48 h,分别采用MTT法检测细胞增殖,使用流式细胞仪检测细胞凋亡,采用Western blot法检测凋亡相关蛋白(Bax、Bcl-2、Caspase-3)、细胞自噬相关蛋白(LC3-I、LC3-II、p62)和PI3K/Akt/NF-κB通路蛋白表达情况。结果自小剂量、中剂量到大剂量,紫草素处理HepG2细胞增殖抑制率分别为(23.7±3.5)%、(36.2±6.1)%和(56.9±8.3)%,均显著高于对照组[(0.0±0.0)%,P<0.05],细胞凋亡率分别为(19.2±5.3)%、(37.4±7.6)%和(58.6±8.8)%,均显著高于对照组[(2.5±1.2)%,P<0.05];细胞凋亡相关蛋白表达Bax/Bcl-2比值和Caspase-3相对表达量分别为(1.3±0.2)和(2.7±0.3)、(8.2±0.6)和(0.45±0.10)和(0.78±0.16)和(0.95±0.21),显著高于对照组[分别为(0.6±0.1)和(0.18±0.06),P<0.05];细胞自噬相关蛋白表达LC3-II/LC3-I比值为(1.25±0.08)、(1.43±0.10)和(1.76±0.22),显著高于对照组[(0.96±0.08),P<0.05],p62相对表达水平分别为(0.81±0.09)、(0.62±0.15)和(0.43±0.08),显著低于对照组[(1.06±0.05),P<0.05]; PI3K、Akt和p65蛋白表达水平分别为【(0.64±0.16)、(0.51±0.12)和(0.32±0.06)】、(【0.54±0.17)、(0.37±0.05)和(0.05±0.01)】和【(0.63±0.15)、(0.52±0.10)和(0.36±0.09)],均显著低于对照组[分别为(0.84±0.13)、(0.76±0.15)和(0.89±0.11),P<0.05]。结论紫草素可能通过抑制PI3K/Akt/NF-κB信号通路关键蛋白表达促进HepG2细胞凋亡和自噬,从而具有抑瘤作用。

关键词: HepG2细胞, 紫草素, 磷脂酰肌醇3-激酶, 蛋白激酶, 核转录因子-κB, 体外

Abstract: Objective The aim of this study was to investigate the effects of shikonin, a herbal medicine, on proliferation,apoptosis and phosphatidylinositol 3-kinase (PI3K)/protein kinase (Akt)/nuclear transcription factor-κB (NF-κB) signaling pathway protein expression in HepG2 cells in vitro. Methods HepG2 cells in logarithmic growth phase were treated with different concentration [0(control), 1, 2.5 and 5 μmol/L of shikonin for 48h. The proliferation inhibition rate of HepG2 cells was detected by MTT assay, the apoptosis by flow cytometry and the expression of apoptosis-related proteins (Bax, Bcl-2, Caspase-3), autophagy-related proteins (LC3-I, LC3-II, p62) and PI3K/Akt/NF-κB proteins were detected by Western bloting. Results After intervention for 48 h, the proliferation inhibition rates of cells from low, middle and high dose of shikonin were (23.7±3.5)%, (36.2±6.1)% and (56.9±8.3)%, all significantly higher than [(0.0±0.0)%, P<0.05]in the control, and theapoptosis rates were (19.2±5.3)%, (37.4±7.6)% and (58.6±8.8)%, also significantly higher than [(2.5±1.2)%, P<0.05]in the control; the apoptosis-related protein expression Bax/Bcl-2 ratio and relative expression of Caspase-3 were (1.3±0.2) and (2.7±0.3), (8.2±0.6) and (0.45±0.10), and (0.78±0.16) and (0.95±0.21), all significantly higher than [(0.6±0.1) and (0.18±0.06), respectively, P<0.05]in the control; the autophagy-related protein expression LC3-II/LC3-I ratio were (1.25±0.08), (1.43±0.10) and (1.76±0.22), significantly higher than [(0.96±0.08), P<0.05]in the control, while the relative expression of p62 were (0.81±0.09), (0.62±0.15), (0.43±0.08), significantly lower than that [(1.06±0.05), P<0.05]in the control, and the PI3K, Akt and p65 protein expression were [0.64±0.16), (0.51±0.12) and (0.32±0.06)], [(0.54±0.17), (0.37±0.05) and (0.05±0.01), and [(0.63±0.15), (0.52±0.10) and (0.36±0.09)], all significantly lower than [(0.84±0.13), 0.76±0.15) and (0.89±0.11), respectively, P<0.05]in the control. Conclusion Shikonin promotes apoptosis and autophagy of HepG2 cells in vitro, which might be related to the inhibition of PI3K/Akt/NF-κB signaling pathway protein expression.

Key words: HepG2 cells, Shikonin, phosphatidylinositol 3-kinase, Protein kinase, Nuclear transcription factor-κB, In vitro

马艳华, 黄芬, 王文尖. 紫草素对HepG2细胞增殖、凋亡和PI3K/Akt/NF-κB信号通路蛋白表达的影响^*[J]. 实用肝脏病杂志, 2020, 23(4): 471-475.

Ma Yanhua, Huang Fen, Wang Wenjian. Effects of shikonin on proliferation,apoptosis and PI3K/Akt/NF-κB signaling pathway protein expression in HepG2 cells in vitro[J]. Journal of Practical Hepatology, 2020, 23(4): 471-475.

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紫草素对HepG2细胞增殖、凋亡和PI3K/Akt/NF-κB信号通路蛋白表达的影响^*

Effects of shikonin on proliferation,apoptosis and PI3K/Akt/NF-κB signaling pathway protein expression in HepG2 cells in vitro

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