硫酸小檗碱对猪轮状病毒感染的IPEC-J2中TGF-β_1、EGFR、EGF、IGF_1 mRNA表达的影响  

Berberine Sulfate Affects TGF-β_1,EGFR,EGFand IGF_1 mRNA Expression in Rotavirus-infected IPEC-J2

作　　者：徐环叶 李妹玲 陈立功[1,2] 王丽叶 孟利佳 崔欢 张诚 白晓 王迎春 董世山 XU Huan-ye;LI Mei-ling;CHEN Li-gong;WANG Li-ye;MENG Li-jia;CUI Huan;ZHANG Cheng;BAI Xiao;WANG Ying-chun;DONG Shi-shan(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;North China Research Center of Animal Epidemic Pathogen Biology of China Agriculture Ministry,Baoding 071001,China;Handan Agriculture and Animal Husbandry Bureau,Handan 056400,China)

机构地区：[1]河北农业大学动物医学院,保定071001 [2]农业部动物疫病病原生物学华北科学观测实验站,保定071001 [3]邯郸市涉县农牧局,邯郸056400

出　　处：《畜牧兽医学报》2018年第10期2283-2291,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31470125);河北农业大学科研发展基金(1699003)

摘　　要：利用猪轮状病毒(PoRV)感染仔猪肠黏膜上皮细胞(IPEC-J2)建立损伤模型,研究硫酸小檗碱(BS)对PoRV感染的IPEC-J2中TGF-β_1、EGFR、EGF、IGF_1基因mRNA表达的影响。通过构建目的基因和内参基因的重组质粒,绘制荧光标准曲线,采用实时荧光定量PCR法测定BS对PoRV感染的IPEC-J2中TGF-β_1、EGFR、EGF、IGF_1mRNA表达的影响。结果显示:PoRV能够显著引起IPEC-J2变性和坏死脱落,用BS药物组的IPEC-J2则生长基本良好;同时计算试验组相对于对照组各因子mRNA表达的差异倍数,显示BS有促进病毒损伤IPEC-J2中TGF-β_1和EGFR mRNA表达,抑制EGF和IGF_1mRNA表达的作用。可见在肠黏膜细胞损伤修复过程中,BS能够显著促进TGF-β_1和EGFR基因表达,但是抑制EGF和IGF_1基因表达,表明BS对各细胞因子在修复过程中发挥的作用和时间存在明显差异。BS可能通过调节损伤的肠黏膜上皮细胞修复因子的基因表达,进而调控肠上皮细胞的增殖、转化,对受损肠黏膜起到保护和修复作用。This study was conducted to investigate the effects of berberine sulfate(BS)on TGF-β1,EGFR,EGF and IGF 1 expression in porcine intestinal epithelial cell(IPEC-J2)that were infected by rotavirus.We established a model in which IPEC-J2 were infected with rotavirus.We established target and reference gene recombinant plasmid and standard curve.TGF-β1,EGFR,EGF and IGF 1 mRNA levels were quantified in BS and rotavirus treated IPEC-J2 by real-time polymerase chain reaction.Results were as follows:after the infection of control IPEC-J2 with rotavirus,the cells clustered into poorly-organized groups,shed or died.By contrast,the BS-treated IPEC-J2 that were infected with rotavirus formed highly-organized groups.The difference of TGF-β1,EGFR,EGF and IGF 1 gene mRNA expression in each experimental group was compared with that of the control group,showing that BS can promote the expression of TGF-β1 and EGFR.However,BS inhibited the expression of EGF and IGF 1 mRNA.Compared with EGF and IGF 1 gene expression decreased significantly,the expression of TGF-β1 and EGFR gene were increased significantly,suggesting that the effect of repair factors may not be the same in the process of repairing cells injury.Our results show that BS could ameliorate IPEC-J2 injury by regulating repair factors expression to promote the proliferation and transformation of cells for repairing the intestinal mucosa.

关 键 词：硫酸小檗碱 仔猪肠黏膜上皮细胞 猪轮状病毒 实时荧光定量PCR TGF-β1 EGFR EGF IGF1 

分 类 号：S853.7[农业科学—临床兽医学]