丁酸钠和吲哚-3-丙酸共处理对Caco-2细胞紧密连接以及炎症因子的影响  

Effects of Co-Treatment of Sodium Butyrate and Indole-3-Propionic Acid on Tight Junctions and Inflammatory Cytokines in Caco-2 Cells

作　　者：王轲信 吴娴 龚海洲 方巧玉 李向臣 张亚南 WANG Kexin;WU Xian;GONG Haizhou;FANG Qiaoyu;LI Xiangchen;ZHANG Yanan(Zhejiang Provincial Engineering Laboratory for Animal Health Inspection&Internet Technology,Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province,College of Animal Science and Technology,College of Veterinary Medicine,Zhejiang Agricultural and Forestry A&F University,Hangzhou 311300,China;The Key Laboratory of Molecular Animal Nutrition,Ministry of Education,College of Animal Science,Zhejiang University,Hangzhou 310058,China;College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]浙江农林大学动物科技学院·动物医学院浙江省畜禽绿色生态健康养殖应用技术研究重点实验室动物健康互联网检测技术浙江省工程实验室,杭州311300 [2]浙江大学动物科学学院动物分子营养学教育部重点实验室,杭州310058 [3]扬州大学动物科学与技术学院,扬州225009

出　　处：《畜牧兽医学报》2024年第11期5124-5134,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：浙江农林大学科研发展基金项目(2023LFR006);浙江农林大学国家级大学生创新创业训练计划(202310341010,张亚南);浙江大学动物分子营养学教育部重点实验室开放性课题(KLMAN202307)。

摘　　要：本研究以人克隆结肠腺癌Caco-2细胞系为模型,旨在探究丁酸钠(sodium butyrate,NaB)和吲哚-3-丙酸(indole-3-propionic acid,IPA)共处理对Caco-2细胞紧密连接和炎症因子的影响。使用0.4 mmol·L^(-1)NaB和0.1 mmol·L^(-1)IPA分别和共处理Caco-2细胞24 h,检测细胞跨膜电阻(TEER)值、细胞紧密连接蛋白表达、炎症因子的基因表达和细胞上清液中炎症因子的分泌水平。结果发现,与对照组相比,NaB和IPA共处理组均显著提高TEER值(P<0.01),而NaB和IPA单独处理组TEER值无显著变化(P>0.05)。qPCR和蛋白免疫印迹结果表明,与对照组相比,共处理组显著下调了Claudin-2的mRNA相对表达量(P<0.01);与NaB单独处理组相比,共处理组显著上调了Claudin-1的mRNA相对表达量(P<0.05),并且极显著促进了Claudin-1、ZO-1和Occludin蛋白的表达(P<0.001);与IPA单独处理组相比,共处理组显著上调了ZO-1的mRNA相对表达量(P<0.05),并极显著提高了Claudin-1和ZO-1蛋白表达量(P<0.001)。在炎症因子方面,与对照组相比,NaB和IPA共处理组显著下调炎症因子IL-6、IL^(-1)β、IL-8及TNF-α的mRNA相对表达量(P<0.05)。ELISA结果显示,与对照组相比,共处理极显著降低了细胞上清液中IL-6、IL^(-1)β、IL-8及TNF-α的分泌量(P<0.001)。本研究表明,相比于NaB或IPA单独处理,NaB和IPA共处理可增加细胞跨膜电阻值,促进细胞间紧密连接蛋白表达并且降低促炎因子的分泌,降低了肠道通透性。综上,NaB和IPA共处理在增强上皮细胞屏障功能以及降低炎症反应方面具有更强的作用。This study aims to investigate the effects of co-treatment of sodium butyrate and indole-3-propionic acid on the expression of tight junction-related proteins and inflammatory factors in Caco-2 cells.Caco-2 cells were administrated with 0.4 mmol·L^(-1)sodium butyrate and 0.1 mmol·L^(-1)indole-3-propionic acid each and together for 24 h;Then the transmembrane resistance(TEER)value,expression of tight junction related proteins,gene expression and secretion level of inflammatory cytokines were measured.The results showed that,compared with control group,the co-treatment of sodium butyrate and indole-3-propionic acid significantly increased the TEER value in Caco-2 cells(P<0.01),while no significant changes were observed in sodium butyrate or indole-3-propionic group(P>0.05);In addition,the results by qPCR and Western blot showed that the relative expression of Claudin-2 mRNA was significantly down-regulated in the co-treatment group compared with the control group(P<0.01);Compared with sodium butyrate group,the relative mRNA expression of Claudin-1 was significantly up-regulated in the co-treatment group(P<0.05),and the protein expression of Claudin-1,ZO-1 and Occludin was highly significantly increased in the co-treatment group(P<0.001);Compared with the indole-3-propionic acid group,the relative mRNA expression of ZO-1 was significantly up-regulated in the co-treatment group(P<0.05),and the protein expression of Claudin-1 and ZO-1 was highly significantly increased(P<0.001).For inflammatory cytokines,the relative mRNA expression levels of IL-6,IL^(-1)β,IL-8 and TNF-αwere significantly down-regulated in the co-treatment group(P<0.05);Meanwhile,the results of ELISA showed that the co-treatment group highly significantly suppressed the secretion of IL-6,IL^(-1)β,IL-8 and TNF-αin the cell supernatant compared with the control group(P<0.001).In conclusion,compared with sodium butyrate or indole-3-propionic acid,the co-treatment of sodium butyrate and indole-3-propionic acid increased the TEER value,promoted the e

关 键 词：丁酸钠 吲哚-3-丙酸 紧密连接 炎症因子 CACO-2细胞 

分 类 号：S816[农业科学—饲料科学]