肿瘤坏死因子-α刺激对IPEC-1细胞活力、物理屏障以及炎症相关基因表达的影响  

Effects of Tumor Necrosis Factor-αStimulation on Viability,Physical Barrier and Inflammation-Related Gene Expression of IPEC-1 Cells

作　　者：李佩 王树辉[1] 陈少魁 王秀英[1] 肖勘 刘玉兰[1] 王丹 LI Pei;WANG Shuhui;CHEN Shaokui;WANG Xiuying;XIAO Kan;LIU Yulan;WANG Dan(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》2020年第9期4337-4344,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：湖北省自然科学基金创新群体项目“细胞死亡与猪肠道疾病的关系及其营养调控”(2019CFA015)。

摘　　要：本试验探讨了肿瘤坏死因子-α(TNF-α)刺激对IPEC-1细胞活力、乳酸脱氢酶(LDH)活性、紧密连接蛋白和炎症相关基因表达的影响。分别选取0(对照组)、10、20、40、50 ng/mL的TNF-α刺激IPEC-1细胞12、24、48 h,检测相关指标。结果表明:1)与对照组相比,当刺激时间为24 h时,不同浓度的TNF-α均导致细胞活力显著下降(P<0.05);当刺激时间为48 h时,40和50 ng/mL的TNF-α导致细胞活力显著下降(P<0.05)。2)与对照组相比,当刺激时间为24和48 h时,40和50 ng/mL的TNF-α导致LDH活性显著升高(P<0.05)。3)与对照组相比,当刺激时间为24 h时,40和50 ng/mL的TNF-α导致IPEC-1细胞紧密连接蛋白闭合小环蛋白-1(ZO-1)、闭合蛋白-1(Claudin-1)、咬合蛋白(Occludin)的mRNA表达量显著上升(P<0.05);当刺激时间为48 h时,不同浓度的TNF-α对Claudin-1蛋白表达量无显著影响(P>0.05)。4)与对照组相比,当刺激时间为24和48 h时,不同浓度的TNF-α均导致TNF-α和白细胞介素-8(IL-8)的mRNA表达量显著上升(P<0.05),并呈时间和剂量依赖关系。以上结果表明,TNF-α刺激诱导了IPEC-1细胞的炎症反应,引起了IPEC-1细胞损伤。综合考虑各指标,可采用40 ng/mL TNF-α刺激IPEC-1细胞48 h构建IPEC-1细胞损伤模型。This study was investigated the effects of tumor necrosis factor-α(TNF-α)stimulation on viability,lactate dehydrogenase(LDH)activity,tight junction protein and inflammation-related gene expression of IPEC-1 cells.Selected 0(control group),10,20,40 and 50 ng/mL of TNF-αto stimulate IPEC-1 cells for 12,24 and 48 h,respectively,and detected relevant indexes.The results showed as follows:1)compared with the control group,different concentrations of TNF-αsignificantly decreased cell viability at stimulation for 24 h(P<0.05);TNF-αat 40 and 50 ng/mL resulted in a significant decrease in cell viability at stimulation for 48 h(P<0.05).2)Compared with the control group at stimulation for 24 and 48 h,TNF-αat 40 and 50 ng/mL led to a significant increase in LDH activity(P<0.05).3)Compared with the control group at stimulation for 24 h,TNF-αat 40 and 50 ng/mL significantly increased the mRNA expression levels of tight junction proteins zonula occluden-1(ZO-1),Claudin-1 and Occludin of IPEC-1 cells(P<0.05);different concentrations of TNF-αhad no significant effects on Claudin-1 protein expression level at stimulation for 48 h(P>0.05).4)Compared with the control group at stimulation for 24 and 48 h,different concentrations of TNF-αsignificantly increased the mRNA expression levels of TNF-αand interleukin-8(IL-8)(P<0.05),and in a time-and dose-dependent manner.The above results indicate that TNF-αstimulation induces inflammatory response and damage in IPEC-1 cells.By comprehensive consideration,the IPEC-1 cell damage model can be induced by 40 ng/mL TNF-αat stimulation for 48 h.

关 键 词：IPEC-1细胞 TNF-Α 细胞活力 LDH活性 紧密连接蛋白 炎症相关基因 

分 类 号：S811.3[农业科学—畜牧学]