Zhang Weiping, Chen Lihong, Shen Yuxian, Xu Jianming
Department of Gastroenterology, The First Affiliated Hospital of Anhui Medical University, Key Laboratory of Gastroenterology of Anhui Province, China; The First Affiliated Hospital of AUTCM, Anhui, China.
Department of Gastroenterology, The First Affiliated Hospital of Anhui Medical University, Key Laboratory of Gastroenterology of Anhui Province, China.
Toxicol In Vitro. 2016 Oct;36:186-196. doi: 10.1016/j.tiv.2016.07.017. Epub 2016 Jul 26.
Endoplasmic reticulum (ER) stress-induced cell injury plays an important role in the development of drug-induced liver injury (DILI). However, little is known about the contribution of ER stress to RFP-induced cell injury. In our study, L02 cells were treated with different concentrations of RFP for different time intervals, and cell apoptosis, the survival rate, and the gene and protein expression of GRP78, PERK, ATF4, and CHOP were measured. Additionally, L02 cells were transfected with CHOP-siRNA or a CHOP-over expression plasmid or administered 4-PBA before treatment with RFP. We found that RFP increased the cell apoptosis rate, decreased cell survival, and increased the protein and gene levels of GRP78, PERK, ATF4 and CHOP in both a dose-dependent and a time-dependent manner. Following the transient knockdown of CHOP and treatment with RFP, cell apoptosis decreased and the survival rate increased. Overexpression of CHOP produced the opposite effects. Treatment with 4-PBA decreased the protein and gene expression of GRP78, PERK, ATF4 and CHOP. Additionally, 4-PBA reduced cell apoptosis, increased cell survival and decreased the level of ALT, AST, AKP, LDH and ATP in the cell culture supernatant. These results indicate that 4-PBA alleviates RFP-induced injury in L02 cells via inhibition of the PERK-ATF4-CHOP pathway.
内质网（ER）应激诱导的细胞损伤在药物性肝损伤（DILI）的发生发展中起重要作用。然而，关于内质网应激对RFP诱导的细胞损伤的作用知之甚少。在我们的研究中，用不同浓度的RFP处理L02细胞不同时间间隔，检测细胞凋亡、存活率以及GRP78、PERK、ATF4和CHOP的基因和蛋白表达。此外，在用RFP处理前，将L02细胞转染CHOP-siRNA或CHOP过表达质粒或给予4-PBA。我们发现RFP以剂量和时间依赖性方式增加细胞凋亡率、降低细胞存活率，并增加GRP78、PERK、ATF4和CHOP的蛋白和基因水平。CHOP短暂敲低并经RFP处理后，细胞凋亡减少，存活率增加。CHOP过表达产生相反的效果。4-PBA处理降低了GRP78、PERK、ATF4和CHOP的蛋白和基因表达。此外，4-PBA减少细胞凋亡，增加细胞存活率，并降低细胞培养上清液中ALT、AST、AKP、LDH和ATP的水平。这些结果表明，4-PBA通过抑制PERK-ATF4-CHOP途径减轻RFP诱导的L02细胞损伤。
