Effect of TBBPA-DHEE exposure on rat adrenal pheochromocytoma PC12 cells and its mechanism
ZENG Zhengjia1, FENG Weiwei1, DING Yangyang2, QIAN Xian1, LUO Mengna1, MAO Guanghua1, YANG Liuqing3, WU Xiangyang1
1. School of the Environment and Safety Engineering, 2. Laboratory Animal Research Center, 3. School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang Jiangsu 212013, China
Abstract：Objective： To study the effect of tetrabromobisphenol A bis (2-hydroxyetyl) ether (TBBPA-DHEE) on rat adrenal pheochromocytoma PC12 cells and mechanisms. Methods： PC12 cells were randomly divided into 5 groups: control group, solvent control group (0.05% dimethyl sulfoxide), TBBPA-DHEE low-dose group (5 μg/mL), medium-dose group (20 μg/mL) and high-dose group (35 μg/mL). Experimental exposure lasted for 48 hours according to corresponding groups. MTT assay was used to analyze the half maximal inhibitory concentration (IC50) of TBBPA-DHEE on PC12 cells. The activity of superoxide dismutase (SOD), and contents of malondialdehyde (MDA) and reactive oxygen species (ROS) in PC12 cells were detected by corresponding assay kits. In addition, the contents of nitric oxide (NO) and Ca2+ in cell culture medium were also detected. The content of PC12 cells voltage-gated calcium channel (VGCC) was determined by ELISA method. The expression of proteins in calcium signaling pathway was determined by Western blotting. Results： TBBPA-DHEE (≥20 μg/mL) exposure significantly inhibited the viability of PC12 cells, with IC50 of 33.4 μg/mL. Compared with the control group, 5, 20 and 35 μg/mL TBBPA-DHEE groups PC12 cells SOD activity, MDA and ROS contents were significantly increased (P<0.05), the contents of NO and Ca2+ in the cell culture medium were significantly increased (P<0.01 or P<0.05), and the content of VGCC was markedly decreased (P<0.01); the relative expression levels of p-CaMKⅡ, p-ERk1/2 and p-CREB proteins in calcium ion signaling pathway of 5 μg/mL TBBPA-DHEE group were remarkably decreased (P<0.01); the relative expression levels of CaM, MKP-1, p-CaMKⅡ, p-ERk1/2 and p-CREB proteins in 20 and 35 μg/mL TBBPA-DHEE groups were significantly decreased (P<0.01). Conclusion: Exposure to TBBPA-DHEE could significantly inhibit the viability of PC12 cells and lead to cellular oxidative damage. TBBPA-DHEE may cause calcium dyshomeostasis by down-regulating the expression of VGCC, and then disrupt the expression of calcium signaling pathway related proteins, thus producing neurotoxicity.
［Key words］tetrabromobisphenol A bis (2-hydroxyetyl) ether (TBBPA-DHEE); rat adrenal pheochromocytoma cell (PC12); voltage-gated calcium channel (VGCC); oxidative stress; neurotoxicity
基金资助:国家自然科学基金面上项目(21976072)
通讯作者:
吴向阳，教授，博士生导师，E-mail： wuxy@ujs.edu.cn
作者简介: 曾政嘉(1997—)，男，硕士研究生
引用本文:
曾政嘉， 冯伟伟， 丁阳阳，等. TBBPA-DHEE暴露对大鼠肾上腺髓质嗜铬细胞瘤PC12细胞的影响及机制[J]. 江苏大学学报:医学版, 2023, 33(03): 225-230，244.
ZENG Zhengjia, FENG Weiwei, DING Yangyang, et al. Effect of TBBPA-DHEE exposure on rat adrenal pheochromocytoma PC12 cells and its mechanism. Journal of Jiangsu University(Medicine Edition), 2023, 33(03): 225-230，244.
