LPS通过ROS/p38/MAPK信号通路对鹅胚肝细胞凋亡的影响

doi:10.16431/j.cnki.1671-7236.2023.06.006

Abstract

Abstract: 【Objective】 This study was aimed to establish the isolation and culture technology of goose embryo-derived hepatocytes, explore the mechanism of lipopolysaccharide (LPS) affecting the apoptosis of goose embryo-derived hepatocytes, and provide a reference basis for the study of liver function in goose.【Method】 16-18 days old goose embryos without specific pathogens were selected, and the livers of goose embryos were taken under sterile conditions and digested with 2% collagenase Ⅳ.After passing through the cell sieve, the goose embryo-derived hepatocytes were isolated, cultured and identified by PAS staining.Different concentrations (0 (control group), 0.1, 1.0 and 10 μg/mL) of LPS were added, and the cells were collected at 12, 24 and 36 h, respectively.The level of reactive oxygen species (ROS) and apoptosis in goose embryo-derived hepatocytes were detected by a kit and flow cytometry, respectively.The expression of key apoptosis genes caspase3, caspase9 and p38 mRNA in goose embryo-derived hepatocytes were detected using Real-time quantitative PCR.【Result】 Hepatocytes from goose embryo were successfully isolated and cultured, which had complete morphology, high adhesion rate, high survival rate and stable growth.The results of PAS staining showed that the goose embryo-derived hepatocytes was purplish red in various shades and the nucleus of liver was blue.After adding different concentrations of LPS, oxidative damage was found in goose embryo-derived hepatocytes.When LPS level was 1.0 μg/mL, the level of ROS in goose embryo-derived hepatocytes increased.The late apoptosis of goose embryo-derived hepatocytes was inhibited after the addition of LPS.Except for 36 h, the expression of the key apoptosis genes caspase3, caspase9 and p38 mRNA in goose embryo-derived hepatocytes were significantly decreased (P<0.05).【Conclusion】 In this study, a relatively stable method for isolation and culture of goose embryo-derived hepatocytes was established.After adding different concentrations of LPS, the level of ROS in goose embryo-derived hepatocytes was increased, p38/MAPK pathway was activated, and apoptosis was inhibited.

Key words: goose embryo-derived hepatocytes; lipopolysaccharide (LPS); ROS/p38/MAPK signaling pathway; apoptosis

CLC Number:

S852.2

ZHONG Yueyun, YANG Shuzhan, CHEN Feiyue, LU Zhier, LI Bingxin, LI Wanyan, TIAN Yunbo, HUANG Yunmao, XU Danning, CAO Nan. Effects of LPS on Apoptosis of Goose Embryo-derived Hepatocytes ROS/p38/MAPK Signaling Pathway[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2224-2232.

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Effects of LPS on Apoptosis of Goose Embryo-derived Hepatocytes ROS/p38/MAPK Signaling Pathway

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