APS对DEX诱导的IPEC-J2细胞凋亡的保护作用  

Protective effects of APS against DEX-induced apoptosis in IPEC-J2 cells

作　　者：谢维娜 田朋 单兴根 丁红研[1] 王蕾 赵畅 李玉[1] 冯士彬 王希春[1] 吴金节[1] XIE Wei-na;TIAN Peng;SHAN Xing-gen;DING Hong-yan;WANG Lei;ZHAO Chang;LI Yu;FENG Shi-bin;WANG Xi-chun;WU Jin-jie(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China)

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《中国兽医科学》2023年第1期127-134,共8页Chinese Veterinary Science

基　　金：安徽省生猪产业技术体系资助项目(AHCYJSTX-05-03)。

摘　　要：本试验旨在探讨黄芪多糖(APS)对地塞米松(DEX)诱导的猪小肠上皮细胞(IPEC-J2)凋亡的保护作用。采用不同浓度的APS(0~0.8 mg/m L)或DEX(0~0.7 mg/m L)处理IPEC-J2细胞24 h,利用细胞计数试剂(CCK-8)检测细胞活性,确定药物作用浓度。对照组无添加,A组添加0.4 mg/m L APS培养,D组添加0.4 mg/m L DEX培养,A+D组添加0.4 mg/m L APS+0.4 mg/m L DEX共培养。采用Calcein/PI细胞活性与细胞毒性检测试剂盒检测细胞的存活情况,流式细胞仪检测IPEC-J2细胞的凋亡情况,透射电子显微镜观察细胞的形态,实时荧光定量PCR检测凋亡基因Caspase3、Caspase9、Bid、Bcl-2 m RNA的相对表达量,Western-blotting检测Caspase3、Caspase9、Bid、Bcl-2蛋白的表达水平。结果表明,APS可以通过调控凋亡蛋白和基因的表达来抑制DEX诱导的IPEC-J2细胞的凋亡,恢复IPEC-J2细胞的活性,起到保护猪小肠上皮细胞作用。The purpose of this experiment was to investigate the protective effect of Astragalus polysaccharide(APS)against dexamethasone(DEX)-induced apoptosis in IPEC-J2 cells.IPEC-J2 cells were treated with different concentrations of APS(0-0.8 mg/m L)or DEX(0-0.7 mg/m L)for 24 h,and the cell activity was determined by the cell counting reagent(CCK-8)to determine the concentration of drug action.Without addition in control group,group A was supplemented with 0.4 mg/m L APS,group D with 0.4mg/m L DEX,and group A+D with both 0.4 mg/m L APS and 0.4 mg/m L DEX.Cell survival was measured by the Calcein/PI cell activity and cytotoxicity detection kit,apoptosis of IPEC-J2 cells was measured by flow cytometry,the morphology of the cells was observed by a transmission electron microscope,the relative expression amount of the apoptotic genes Caspase3,Caspase9,Bid,and Bcl-2 m RNA was measured by quantitative real-time PCR,and the expression levels of Caspase3,Caspase9,Bid,and Bcl-2 proteins were determined by Western-blot.The results showed that,APS can inhibit the DEX-induced apoptosis of IPEC-J2 cells by regulating the expression of apoptotic proteins and genes,restore the activity of IPEC-J2 cells,and protect intestinal epithelial cells.

关 键 词：黄芪多糖 地塞米松 猪小肠上皮细胞 细胞凋亡 

分 类 号：S853.74[农业科学—临床兽医学]