ALOX15B-JNK在热应激诱导支持细胞氧化应激和凋亡中的作用

doi:10.11843/j.issn.0366-6964.2023.12.016

Abstract

Abstract: The objective of this study was to investigate whether arachidonate lipoxygenase type 15B (ALOX15B) participates in the production of malondialdehyde (MDA) as well as reactive oxygen species (ROS), regulates the apoptosis of porcine sertoli cells and determines the role of JNK in the process. In this experiment, porcine sertoli cells from the testicular tissues of 20 3-week-old three-way hybrid piglets was isolated and randomly divided into 3 groups with 3 replicates in each group. Porcine sertoli cells were treated at 44℃ for 30 min as the heat stress model. After siRNA transfection of ALOX15B and addition of ALOX15B inhabitor (baicalein) or JNK inhibitor, the apoptosis rates of the cells were detected by flow cytometry, the expressions of phosphorylated JNK, ALOX15B and p53 were detected by Western blotting, and the contents of 8-HETE and 15-HETE were analyzed by ELISA. The levels of MDA and ROS were detected by the kit. The results showed that the combined treatment of baicalein and heat stress decreased the content of 8-HETE and 15-HETE (P<0.01), reduced the levels of MDA and ROS(P<0.01), and inhibited the apoptosis rate induced by heat stress (P<0.01). Under heat stress, bai-calein and ALOX15B siRNA inhibited JNK phosphorylation induced by heat stress (P<0.01). SP600125 inhibited the expression of p53 downstream by decreasing the phosphorylation of JNK (P<0.01), and reduced the apoptosis induced by heat stress (P<0.01). Meanwhile, compared to the heat stress group, SP600125 combined with heat stress also gave negative feedback on the protein level of ALOX15B (P<0.01), decreased the content of lipid peroxides 8-HETE (P<0.01) and 15-HETE (P<0.05), and reduced the level of MDA and ROS (P<0.01). In conclusion, ALOX15B regulates apoptosis of porcine sertoli cells under heat stress; JNK participated in the effect of ALOX15B, and regulated apoptosis of porcine sertoli cells through p53, and could have the feedback on the levels of ALOX15B, 8-HETE, 15-HETE, MDA and ROS.

Key words: heat stress, sertoli cells, JNK, apoptosis, ALOX15B

CLC Number:

S828.3

XUE Hongyan, YANG Mengyu, YANG Huan, DONG Lijun, CAI Xiaqing, ZHAO Zemin, WANG Xianzhong. The Role of ALOX15B-JNK in Heat Stress-induced Oxidative Stress and Apoptosis of Sertoli Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5056-5065.

References

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The Role of ALOX15B-JNK in Heat Stress-induced Oxidative Stress and Apoptosis of Sertoli Cells

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