Triptolide-induced HepG2 Cells Hepatotoxicity Through Autophagy

LIU Chun-hui; ZHOU Ling-ling; MA Ke-xun; WANG Jing; FENG Zhe; ZHOU Xue-ping

doi:10.13422/j.cnki.syfjx.2017100099

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Triptolide-induced HepG2 Cells Hepatotoxicity Through Autophagy

更新时间：2024-01-04

- Triptolide-induced HepG2 Cells Hepatotoxicity Through Autophagy
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 10, Pages: 99-103(2017)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2017100099
  CLC： R285.1
- Published：2017
- 稿件说明：
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LIU Chun-hui, ZHOU Ling-ling, MA Ke-xun, et al. Triptolide-induced HepG2 Cells Hepatotoxicity Through Autophagy[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(10): 99-103.
DOI：

LIU Chun-hui, ZHOU Ling-ling, MA Ke-xun, et al. Triptolide-induced HepG2 Cells Hepatotoxicity Through Autophagy[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(10): 99-103. DOI： 10.13422/j.cnki.syfjx.2017100099.

摘要

目的：探讨雷公藤甲素（TP）对HepG2细胞的过氧化损伤及对自噬的调控作用。方法：将质量浓度为0.003 2，0.016，0.08，0.4，2 g·L-1的TP作用于体外培养的HepG2细胞48 h，另设空白组做对比，采用噻唑蓝（MTT）法检测细胞生长活性，采用酶联免疫吸附测定（ELISA）法检测超氧化物歧化酶（SOD），谷胱甘肽过氧化物酶（GSH-Px）的活性，采用蛋白质免疫印迹（Western blot）法检测自噬相关蛋白微管相关蛋白轻链3（LC3Ⅱ）与Beclin1蛋白的表达情况，用免疫荧光法检测自噬相关蛋白LC3的表达情况。结果：与空白组比较，随着TP质量浓度的增加，HepG2活性下降（P<0.05，P<0.01）；检测细胞上清中SOD，GSH-Px的活性，均较空白组降低（P<0.05，P<0.01）；Western blot表明TP可上调LC3Ⅱ与Beclin1蛋白水平的表达（P<0.05，P<0.01），免疫荧光法同样表明TP能够引起自噬相关蛋白LC3表达的增多（P<0.05，P<0.01），均具有一定的浓度依赖性。结论：一定质量浓度的TP可造成肝细胞损伤，其损伤机制可能与过氧化损伤以及过氧化损伤导致的自噬过度激活有关。

Abstract

Objective: To investigate the toxicity of triptolide on HepG2 cells and its modulating effects on autophagy. Method: HepG2 cells were cultured in vitro

and triptolide with the concentrations of 0.003 2

0.016

0.08

0.4

2 g·L-1 were added into the cells for 48 h. Compared with the control group

the cells viability was detected by MTT assay

ELISA methods was used to determine superoxide dismutase (SOD)

glutathion peroxidase (GSH-Px) activity

the expressions of the autophagy-related protein LC3Ⅱ and Beclin1 were detected by Western blot

and the expression of the autophagy-related protein LC3 was detected by immunofluorescence. Result: Compared with the control group

with the increase in the dose of triptolide

the HepG2 activity was decreased (P<0.05

P<0.01); the levels of SOD and GSH-Px expressions were lower than the control group (P<0.05

P<0.01); the expressions of autophagy-related protein LC3Ⅱ and Beclin1 were detected by Western blot (P<0.05

P<0.01)

and the expression of autophagy-related protein LC3 was also detected by immunofluorescence in a dose-dependent manner (P<0.05

P<0.01). Conclusion: Certain concentrations of triptolide could cause hepatotoxicity

which was related to peroxidation and overactivating autophagy caused peroxidation.

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