Fas/FasL通路对镉激活PC12细胞MAPK通路的影响

doi:10.11843/j.issn.0366-6964.2021.07.025

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (7): 2034-2043.doi: 10.11843/j.issn.0366-6964.2021.07.025

Fas/FasL通路对镉激活PC12细胞MAPK通路的影响

王莉^1,2, 陈洁^1,3, 闻双全^1,2, 邹辉^1,2, 顾建红^1,2, 刘学忠^1,2, 卞建春^1,2, 刘宗平^1,2, 袁燕^1,2*

1. 扬州大学兽医学院, 扬州 225009;
2. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009;
3. 南通市动物疫病预防控制中心, 南通 226011

收稿日期:2020-11-17 出版日期:2021-07-23 发布日期:2021-07-23
通讯作者: 袁燕,主要从事动物营养代谢病与中毒病研究,E-mail:yuanyan@yzu.edu.cn
作者简介:王莉(1994-),女,河南汝南人,硕士生,主要从事动物营养代谢病与中毒病研究,E-mail:707361277@qq.com
基金资助:
国家自然科学基金（31772808）；江苏高校优势学科建设工程资助项目（PAPD）；扬州大学优秀青年骨干教师资助项目；扬州大学校教改课题（YZUJX2019-51C）

Effect of Fas/FasL Pathway on MAPK Pathway Activated by Cadmium in PC12 Cells

WANG Li^1,2, CHEN Jie^1,3, WEN Shuangquan^1,2, ZOU Hui^1,2, GU Jianhong^1,2, LIU Xuezhong^1,2, BIAN Jianchun^1,2, LIU Zongping^1,2, YUAN Yan^1,2*

1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;
3. Nantong Animal Disease Prevention and Control Center, Nantong 226011, China

Received:2020-11-17 Online:2021-07-23 Published:2021-07-23

摘要/Abstract

摘要： 旨在研究Fas/FasL通路对镉激活PC12细胞MAPK通路的影响，用10 μmol L^-1醋酸镉（CdAc₂）分别处理插入非特异性序列PC12细胞与Fas基因沉默PC12细胞12 h；用10 μmol·L^-1CdAc₂与40 μmol·L^-1 Z-IETD-FMK （caspase-8特异性抑制剂）单独或联合处理PC12细胞12 h。通过Western blot检测Fas/FasL通路相关蛋白Fas、FasL、Fas相关死亡域蛋白（FADD）、Cleaved caspase-8、死亡结构域相关蛋白（Daxx）、凋亡信号调节激酶1（ASK1）的表达与MAPK通路相关蛋白ERK1/2、JNK1/2、p-ERK1/2、p-JNK1/2的表达；Hoechst33258荧光染色检测细胞凋亡形态学变化，流式细胞术检测细胞凋亡率。结果显示，Fas shRNA慢病毒极显著抑制镉引起的PC12细胞Fas/FasL通路相关蛋白表达量和凋亡率的升高（P<0.01），缓解镉引起的PC12细胞凋亡形态学变化；Fas shRNA与Z-IETD-FMK均能够极显著抑制镉引起的PC12细胞MAPK通路相关蛋白ERK1/2与JNK1/2磷酸化水平升高（P<0.01）。综上表明，Fas/FasL通路调控MAPK通路参与镉致PC12细胞凋亡。

关键词: 镉, PC12细胞, 凋亡, Fas/FasL通路, MAPK通路

Abstract: To study the effect of Fas/FasL pathway on cadmium-activated MAPK pathway in PC12 cells, PC12 cells with non-specific sequences inserted and Fas gene silenced were treated with 10 μmol·L^-1 CdAc₂ respectively for 12 h, PC12 cells were treated with 10 μmol·L^-1CdAc₂ and 40 μmol·L^-1 Z-IETD-FMK (caspase-8 specific inhibitor) alone or in combination for 12 h. The expression of Fas/FasL pathway-related proteins Fas, FasL, Fas-associated death domain protein (FADD), Cleaved caspase-8, Death domain associated protein (Daxx), Apptosis signal-regulating kinase 1 (ASK1) and MAPK pathway-related proteins ERK1/2, JNK1/2, p-ERK1/2, p-JNK1/2 were detected by Western blot. The morphological changes of apoptosis were detected by Hoechst33258 fluorescence staining, and the apoptosis rate was detected by flow cytometry. The results showed that Fas shRNA lentivirus significantly inhibited the increase of Fas/FasL pathway related-proteins expression and apoptosis rate in PC12 cells induced by cadmium (P<0.01), and alleviated the morphological changes of cell apoptosis caused by cadmium in PC12 cells. In addition, Fas shRNA and Z-IETD-FMK could significantly inhibit the increase of cadmium-induced phosphorylation levels of MAPK pathway-related proteins ERK1/2 and JNK1/2 in PC12 cells (P<0.01). Our results indicate that the regulation of Fas/FasL pathway on the MAPK pathway is involved in cadmium-induced apoptosis in PC12 cells.

Key words: cadmium, PC12 cell, apoptosis, Fas/FasL pathway, MAPK pathway

中图分类号:

S856.9

王莉, 陈洁, 闻双全, 邹辉, 顾建红, 刘学忠, 卞建春, 刘宗平, 袁燕. Fas/FasL通路对镉激活PC12细胞MAPK通路的影响[J]. 畜牧兽医学报, 2021, 52(7): 2034-2043.

WANG Li, CHEN Jie, WEN Shuangquan, ZOU Hui, GU Jianhong, LIU Xuezhong, BIAN Jianchun, LIU Zongping, YUAN Yan. Effect of Fas/FasL Pathway on MAPK Pathway Activated by Cadmium in PC12 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 2034-2043.

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Fas/FasL通路对镉激活PC12细胞MAPK通路的影响

Effect of Fas/FasL Pathway on MAPK Pathway Activated by Cadmium in PC12 Cells

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