活化Nrf2缓解热应激致肉鸡心肌细胞氧化损伤的研究

doi:10.16431/j.cnki.1671-7236.2022.09.008

Abstract

Abstract: 【Objective】 The study was aimed to investigate the alleviating effect of nuclear factor erythroid 2-related factor 2 (Nrf2) on oxidative damage induced by heat stress of broiler cardiomyocytes.【Method】 Primary broiler cardiomyocytes were purified through different-speed adherence method and chemical purification method.The cardiomyocytes were randomly divided into control group, heat stress model group (HS group) and Nrf2 activator pre-treatment heat stress group (TBHQ+HS group).The cardiomyocytes of the control group were cultured without any treatment, the cardiomyocytes of the HS group were treated in a high-temperature (43 ℃) incubator for 2 h, the cardiomyocytes of the TBHQ+HS group were pretreated with 50 μmol/L tert-butylhydroquinone (TBHQ) for 12 h and then heat stress treatment was carried out.The relative viability of the cells was measured by MTT method.The activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA) and the activity of lactate dehydrogenase (LDH) in cell culture medium were detected by colorimetry method.Real-time PCR was used to detect the mRNA expression levels of Nrf2, heme oxygenase-1 (HO-1), glutamate cysteine ligase (GCLC) and NAD(P)H:quinone oxidoreductase 1 (NQO1).The expressions of Nrf2 total protein and HO-1 protein were detected by Western blotting.【Result】 Compared with the control group, the morphology of primary cardiomyocytes was changed and vacuolar reticular structure was shown in heat stress group, the relative activity and SOD activity of cardiomyocytes were decreased extremely significantly (P<0.01), the content of MDA was increased extremely significantly (P<0.01), and the activity of LDH in cell culture medium was increased extremely significantly (P<0.01).The difference of NQO1 and GCLC mRNA expression in the downstream of Nrf2/antioxidant response element (ARE) signal pathway was not significant (P>0.05), the expression of Nrf2 and HO-1 mRNA was remarkably increased (P<0.05), but there was no difference in Nrf2 total protein and HO-1 protein expression (P>0.05). In TBHQ pre-treatment heat stress group, compared with the heat stress group, the vacuolar reticular structure reduced in cardiomyocytes and the relative activity of cardiomyocytes was raised significantly (P<0.05), the activity of SOD was increased extremely significantly (P<0.01), the content of MDA was diminished meaningly (P<0.05), the activity of LDH enzyme in cell culture medium was decreased extremely significantly (P<0.01).The expression of NQO1 and GCLC mRNA in the downstream of Nrf2/ARE signal pathway was increased significantly(P<0.05), the expression of Nrf2 mRNA and total protein was increased significantly (P<0.05), besides, the expression of HO-1 mRNA and protein were all raised extremely significantly (P<0.01).【Conclusion】 Activation of Nrf2 could up-regulate the transcription of antioxidant genes and the expression of protein in the downstream of Nrf2 signaling pathway, improve the antioxidant capacity of heat stressed broiler cardiomyocytes, and alleviate the oxidative damage induced by heat stress.It suggested that Nrf2 might be an effective molecular target for broiler cardiomyocytes to resist oxidative damage induced by heat stress.

Key words: heat stress; broilers; cardiomyocytes; Nrf2; HO-1

CLC Number:

Q494

WU Yanan, SHI Yuxiang, ZHANG Yongying, LIU Guanhui, SONG Weiguang, ZHONG Cuihong, WANG Yongxia. Study on Nrf2 Activation Alleviating Heat Stress-Induced Oxidative Damage in Broiler Cardiomyocytes[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3343-3352.

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Study on Nrf2 Activation Alleviating Heat Stress-Induced Oxidative Damage in Broiler Cardiomyocytes

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