Jian Li , Jia-rong Mo , Shi-yu Hu , Xin Dong , Jia-wei Li , Li-yu Yang , Yi-jian Wu
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Our results showed that pretreatment of IPEC-J2 cells with HEP significantly scavenged ROS and reduced LDH and MDA production. HEP also reduced apoptosis and kept polarity of the mitochondrial membrane potential. Moreover, HEP increased the content of caspase-3 and PARP, and protein expression of Bcl-2, while inhibited Bax and Bad and reduced the content of caspase-9 and release of Cyt<img>C. Meanwhile, HEP inhibited the protein expression of TNFR1, FAS, and FASL, and decreased the content of caspase-8. The results indicated that HEP had a protective effect against oxidative stress in IPEC-J2 cells and the underlying mechanism was reducing apoptosis via mitochondrial and death receptor pathways.</p></div>","PeriodicalId":10602,"journal":{"name":"Comparative Biochemistry and Physiology C-toxicology & Pharmacology","volume":null,"pages":null},"PeriodicalIF":3.9000,"publicationDate":"2024-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Effects of Hericium erinaceus polysaccharide in porcine IPEC-J2 intestinal epithelial cells against apoptosis induced by oxidative stress\",\"authors\":\"Jian Li , Jia-rong Mo , Shi-yu Hu , Xin Dong , Jia-wei Li , Li-yu Yang , Yi-jian Wu\",\"doi\":\"10.1016/j.cbpc.2024.109902\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>This study was intended to investigate whether <em>Hericium erinaceus</em> polysaccharides (HEP) prevent oxidative stress and apoptosis of intestinal porcine epithelial cells from jejunum (IPEC-J2 cells) induced by hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). Crude HEP were extracted and purified by chromatography. The ultraviolet and infrared spectra and monosaccharide composition of HEP were analyzed. Reactive oxygen species (ROS) generation was quantified by flow cytometry method, and lactate dehydrogenase (LDH) and malondialdehyde (MDA) production were determined by TBARS. Also, apoptosis was analyzed by flow cytometry method and the apoptosis-related regulatory molecules were determined by microplate or western blotting method. Our results showed that pretreatment of IPEC-J2 cells with HEP significantly scavenged ROS and reduced LDH and MDA production. HEP also reduced apoptosis and kept polarity of the mitochondrial membrane potential. Moreover, HEP increased the content of caspase-3 and PARP, and protein expression of Bcl-2, while inhibited Bax and Bad and reduced the content of caspase-9 and release of Cyt<img>C. Meanwhile, HEP inhibited the protein expression of TNFR1, FAS, and FASL, and decreased the content of caspase-8. 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引用次数: 0
摘要
本研究旨在探讨麦角多糖(HEP)是否能防止过氧化氢(H2O2)诱导的猪空肠上皮细胞(IPEC-J2 细胞)氧化应激和凋亡。粗 HEP 经提取和层析纯化。分析了 HEP 的紫外光谱、红外光谱和单糖成分。活性氧(ROS)的生成采用流式细胞仪法进行定量,乳酸脱氢酶(LDH)和丙二醛(MDA)的生成采用 TBARS 法进行测定。此外,还采用流式细胞术分析了细胞凋亡,并采用微孔板或 Western 印迹法测定了与细胞凋亡相关的调控分子。结果表明,用 HEP 预处理 IPEC-J2 细胞能明显清除 ROS,减少 LDH 和 MDA 的产生。HEP 还能减少细胞凋亡,保持线粒体膜电位的极性。此外,HEP 还能增加 caspase-3 和 PARP 的含量以及 Bcl-2 的蛋白表达,抑制 Bax 和 Bad,降低 caspase-9 的含量和 CytC 的释放。同时,HEP能抑制TNFR1、FAS和FASL的蛋白表达,降低caspase-8的含量。结果表明,HEP对IPEC-J2细胞的氧化应激具有保护作用,其基本机制是通过线粒体和死亡受体途径减少细胞凋亡。
Effects of Hericium erinaceus polysaccharide in porcine IPEC-J2 intestinal epithelial cells against apoptosis induced by oxidative stress
This study was intended to investigate whether Hericium erinaceus polysaccharides (HEP) prevent oxidative stress and apoptosis of intestinal porcine epithelial cells from jejunum (IPEC-J2 cells) induced by hydrogen peroxide (H2O2). Crude HEP were extracted and purified by chromatography. The ultraviolet and infrared spectra and monosaccharide composition of HEP were analyzed. Reactive oxygen species (ROS) generation was quantified by flow cytometry method, and lactate dehydrogenase (LDH) and malondialdehyde (MDA) production were determined by TBARS. Also, apoptosis was analyzed by flow cytometry method and the apoptosis-related regulatory molecules were determined by microplate or western blotting method. Our results showed that pretreatment of IPEC-J2 cells with HEP significantly scavenged ROS and reduced LDH and MDA production. HEP also reduced apoptosis and kept polarity of the mitochondrial membrane potential. Moreover, HEP increased the content of caspase-3 and PARP, and protein expression of Bcl-2, while inhibited Bax and Bad and reduced the content of caspase-9 and release of CytC. Meanwhile, HEP inhibited the protein expression of TNFR1, FAS, and FASL, and decreased the content of caspase-8. The results indicated that HEP had a protective effect against oxidative stress in IPEC-J2 cells and the underlying mechanism was reducing apoptosis via mitochondrial and death receptor pathways.
期刊介绍:
Part C: Toxicology and Pharmacology. This journal is concerned with chemical and drug action at different levels of organization, biotransformation of xenobiotics, mechanisms of toxicity, including reactive oxygen species and carcinogenesis, endocrine disruptors, natural products chemistry, and signal transduction with a molecular approach to these fields.