槲皮素对产肠毒素大肠杆菌K88诱导的猪肠上皮细胞凋亡和焦亡信号通路的影响  

Effects of Quercetin on Apoptosis and Pyroptosis Signaling Pathways in Porcine Intestinal Epithelium Cells Induced by Enterotoxigenic Escherichia coli K88

作　　者：徐晓叶 龚晗秋 张敏芳 贺鹏伟 周墨涵 刘玉兰[1] 肖勘 XU Xiaoye;GONG Hanqiu;ZHANG Minfang;HE Pengwei;ZHOU Mohan;LIU Yulan;XIAO Kan(Hubei Key Laboratory of Animal Nutrition and Feed Science,Wuhan Polytechnic University,Wuhan 430023,China)

机构地区：[1]武汉轻工大学,动物营养与饲料科学湖北省重点实验室,武汉430023

出　　处：《动物营养学报》2024年第10期6723-6731,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：武汉市科技局知识创新专项曙光计划项目(2022020801020394)。

摘　　要：本试验旨在研究槲皮素(Que)对产肠毒素大肠杆菌K88(ETEC K88)诱导的猪肠上皮细胞损伤、炎症反应、凋亡和焦亡信号通路的影响。试验以猪肠上皮细胞IPEC-1为研究对象,分为4组:对照组、Que组、ETEC K88组、Que+ETEC K88组。用0或10μmol/L Que处理细胞24 h后,再用磷酸盐缓冲液(PBS)或50倍细胞数的ETEC K88刺激细胞2 h,收集样品测定细胞活力、细胞上清液中乳酸脱氢酶(LDH)活性、细胞中炎性因子含量、细胞凋亡和焦亡信号通路相关基因的mRNA表达水平。结果显示:1)Que显著缓解了ETEC K88诱导的细胞活力的下降和细胞上清液中LDH活性的上升(P<0.05)。2)Que显著缓解了ETEC K88诱导的细胞中炎性因子白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)和肿瘤坏死因子-α(TNF-α)含量的上升(P<0.05)。3)Que显著缓解了ETEC K88诱导的细胞凋亡信号通路相关基因凋亡相关因子配体(FasL)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)、B细胞淋巴瘤-2相关X蛋白(Bax)的mRNA表达水平的上升和B细胞淋巴瘤-xL(Bcl-xL)的mRNA表达水平的下降(P<0.05)。4)Que显著缓解了ETEC K88诱导的细胞焦亡信号通路相关基因含pyrin结构域NOD样受体家族3(NLRP3)、IL-18、消皮素D(GSDMD)和NLR家族CARD结构域4(NLRC4)的mRNA表达水平的上升(P<0.05)。综上所述,Que可抑制ETEC K88刺激导致的IPEC-1细胞凋亡和焦亡信号通路的激活,缓解细胞损伤。This study aimed to investigate the effects of quercetin(Que)on injury,inflammatory response,apoptosis and pyroptosis signaling pathways in intestinal porcine epithelial cells induced by enterotoxigenic Escherichia coli(ETEC)K88.Porcine intestinal epithelial cell IPEC-1 was used as the study object,they were divided into four groups:control group,Que group,ETEC K88 group and Que+ETEC K88 group.After trea-ting with 0 or 10μmol/L Que for 24 h,IPEC-1 cells were stimulated with PBS or 50 times cell number of ETEC K88 for 2 h.Samples were collected to measure cell viability,lactate dehydrogenase(LDH)activity in cell supernatant,inflammatory factor contents in cells,and mRNA expression levels of genes related to apopto-sis and pyroptosis signaling pathways.The results showed that:1)Que significantly alleviated the decrease of cell viability and the increase of LDH activity in supernatants induced by ETEC K88(P<0.05).2)Que sig-nificantly relieved the increase of interleukin-1β(IL-1β),interleukin-18(IL-18)and tumor necrosis factor-α(TNF-α)in cells induced by ETEC K88(P<0.05).3)Que significantly alleviated the increase of mRNA ex-pression levels of genes related to apoptosis signaling pathway induced by ETEC K88 such as apoptosis-related factor ligand(FasL),cysteine aspartate protease-3(Caspase-3),cysteine aspartate protease-9(Caspase-9),B-cell lymphoma-2-related X protein(Bax)and the decrease of mRNA expression level of B-cell lymphoma-xL(Bcl-xL)(P<0.05).4)Que significantly relieved the increase of mRNA expression levels of genes related to pyroptosis signaling pathway in cells induced by ETEC K88 such as NOD-like receptors family pyrin domain containing 3(NLRP3),IL-18,gasdermin D(GSDMD)and NLR family CARD domain containing 4(NLRC4)(P<0.05).In summary,Que can inhibit the apoptosis and pyroptosis signaling pathways activated by ETEC K88 in IPEC-1 cells,and alleviate the cell injury.

关 键 词：槲皮素 产肠毒素大肠杆菌K88 IPEC-1 细胞凋亡 细胞焦亡 细胞损伤 

分 类 号：S828[农业科学—畜牧学]