[PERK-eIF2α-ATF4-CHOP通路在双酚A诱导TM4细胞凋亡中的作用]
[The role of PERK-eIF2α-ATF4-CHOP pathway in the apoptosis of TM4 cells induced by bisphenol A].
作者信息
Liu Shuxia, Zhang Ling, Liu Yunhao, Wang Lifang, Quan Chao
机构信息
School of Public Health, Medical College, Hubei Province Key Laboratory of Occupational Hazard Identification and Control, Wuhan University of Science and Technology, Wuhan 430065, China.
School of Public Health, Medical College, Hubei Province Key Laboratory of Occupational Hazard Identification and Control, Wuhan University of Science and Technology, Wuhan 430065, China Central Laboratory, Wuhan Pulmonary Hospital, Wuhan 430030, China.
出版信息
Wei Sheng Yan Jiu. 2023 Jul;52(4):591-597. doi: 10.19813/j.cnki.weishengyanjiu.2023.04.012.
OBJECTIVE
To investigate the effects of bisphenol A(BPA) on the proliferation and apoptosis of mouse testicular sertoli cells(TM4 cells) and the role of PERK-eIF2α-ATF4-CHOP pathway.
METHODS
TM4 cells were treated with different concentrations of BPA(0, 25, 50, 100 μmol/L) and 100 μmol/L BPA combined with protein kinase R-like ER kinase(PERK) inhibitor GSK2656157 for 24 h, and the apoptosis of TM4 cells was observed by TUNEL staining. The expression levels of Bax, Bcl-2, cleaved Caspase-3, GRP78 and PERK-eIF2α-ATF4-CHOP pathway-related proteins were detected by Western blot.
RESULTS
The apoptosis rate of TM4 cells in 25, 50 and 100 μmol/L BPA exposed groups was increased to 3.31%±0.34%, 7.51%±1.10% and 14.58%±0.91%, respectively, which was significantly higher than that in control group(0.73%±0.03%, P<0.05). Compared with the control group(1.00), cleaved Caspase-3 protein expression of TM4 cells in the 25, 50 and 100 μmol/L BPA exposed groups increased to 1.49±0.11, 1.59±0.12, 2.42±0.24, respectively; the ratio of Bax/Bcl-2 increased to 2.06±0.19, 3.94±0.034, 6.14±0.71, respectively; the protein expression of GRP78 increased to 1.29±0.06, 1.39±0.06, 1.92±0.17, respectively; the expression of p-PERK protein was increased to 1.64±0.03, 2.52±0.09, 2.80±0.11, respectively; the expression of p-eIF2α protein was increased to 1.79±0.05, 2.48±0.10, 4.77±0.32, respectively; ATF4 protein expression was increased to 2.51±0.03, 3.24±0.14 and 7.45±0.51, respectively; CHOP protein expression was increased to 1.44±0.01, 3.20±0.11 and 3.80±0.11, respectively, and all the differences were statistically significant(P<0.05). Compared to 100 μmol/L BPA group, the expression level of p-PERK, p-eIF2α, ATF4, CHOP, cleaved Caspase-3 protein and the ratio of Bax/Bcl-2 in 100 μmol/L BPA+10 μmol/L GSK2656157 group were decreased to 2.17±0.11, 1.81±0.13, 1.71±0.23, 2.18±0.22, 1.43±0.03, 2.22±0.13, respectively; the apoptosis rate of TM4 cells was also decreased to 7.28%±0.47%, all the differences were statistically significant(P<0.05).
CONCLUSION
BPA can induce apoptosis of TM4 cells by activating endoplasmic reticulum stress and regulating PERK-eIF2α-ATF4-CHOP pathway.
目的
探讨双酚A（BPA）对小鼠睾丸支持细胞（TM4细胞）增殖和凋亡的影响以及PERK-eIF2α-ATF4-CHOP通路的作用。
方法
用不同浓度的BPA（0、25、50、100μmol/L）及100μmol/L BPA联合蛋白激酶R样内质网激酶（PERK）抑制剂GSK2656157处理TM4细胞24小时，采用TUNEL染色观察TM4细胞凋亡情况。通过蛋白质印迹法检测Bax、Bcl-2、裂解的Caspase-3、GRP78及PERK-eIF2α-ATF4-CHOP通路相关蛋白的表达水平。
结果
25、50和100μmol/L BPA暴露组TM4细胞凋亡率分别升至3.31%±0.34%、7.51%±1.10%和14.58%±0.91%，显著高于对照组（0.73%±0.03%，P<0.05）。与对照组（1.00）相比，25、50和100μmol/L BPA暴露组TM4细胞裂解的Caspase-3蛋白表达分别升至1.49±0.11、1.59±0.12、2.42±0.24；Bax/Bcl-2比值分别升至2.06±0.19、3.94±0.034、6.14±0.71；GRP78蛋白表达分别升至1.29±0.06、1.39±0.06、1.92±0.17；p-PERK蛋白表达分别升至1.64±0.03、2.52±0.09、2.80±0.11；p-eIF2α蛋白表达分别升至1.79±0.05、2.48±0.10、4.77±0.32；ATF4蛋白表达分别升至2.51±0.03、3.24±0.14和7.45±0.51；CHOP蛋白表达分别升至1.44±0.01、3.20±0.11和3.80±0.11，差异均有统计学意义（P<0.05）。与100μmol/L BPA组相比，100μmol/L BPA+10μmol/L GSK2656157组p-PERK、p-eIF2α、ATF4、CHOP、裂解的Caspase-3蛋白表达水平及Bax/Bcl-2比值分别降至2.17±0.11、1.81±0.13、1.71±0.23、2.18±0.22、1.43±0.03、2.22±0.13；TM4细胞凋亡率也降至7.28%±0.47%，差异均有统计学意义（P<0.05）。
结论
BPA可通过激活内质网应激并调节PERK-eIF2α-ATF4-CHOP通路诱导TM4细胞凋亡。
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