猴头菇多糖对氧化应激的IPEC-J2细胞抗氧化能力及紧密连接蛋白ZO-1的影响

doi:10.11843/j.issn.0366-6964.2017.09.024

Abstract

Abstract:

The objective of this study was to evaluate the protective effect of Hericium Erinaceus polysaccharide (HEPs) on intestinal epithelial cell injury induced by H₂O₂. In this study, IPEC-J2 cells were divided into five groups:the control group, the model group, the HEPs low dose treated group, the HEPs medium dose treated group, the HEPs high dose treated group. MTT method was used to select the optimal dose of H₂O₂ and HEPs, respectively; intracellular ROS was observed and determined by DCFH-DA staining; the content of malondialdehyde (MDA) in the supernatant of cultured cells and the activities of superoxide dismutase (SOD), catalase (CAT) and the release rate of lactate dehydrogenase (LDH) in cells were detected by colorimetric method; the transcription level of ZO-1 gene was detected by quantitative real-time PCR (qRT-PCR). The results showed that:(1) The 0.4 mmol·L^-1 was the most suitable H₂O₂ concentration for oxidative stress modeling. (2) The data indicated that H₂O₂ increased the intracellular ROS and induced cell apoptosis significantly, and decreased the vitalities of SOD and CAT significantly; H₂O₂ significantly increased the content of MDA and the release rate of LDH. But, compared with the model group, after HEPs pretreatment, the content of ROS in cells decreased significantly (P<0.01); at the same time, the activities of SOD and CAT were improved significantly (P<0.01), MDA content and LDH release rate were significantly decreased. (3) The results of quantitative real-time PCR displayed that H₂O₂ could significantly reduce the transcription of ZO-1 gene (P<0.01). Compared with H₂O₂ group, HEPs could significantly improve the transcription of ZO-1 genes after HEPs pretreatment (P<0.01). The results of Western blot showed that the expression of ZO-1 protein was similar to that of ZO-1 gene. Results showed that HEPs had protective effect on IPEC-J2 cells in H₂O₂ induced oxidative damage state.

CLC Number:

S856.4

CHEN Xin-yao, ZHANG Jian-long, DONG Xing, CHEN Jing-jie, HUANG Yi-fan, LI Jian. Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1769-1776.

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Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress

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