鼠李糖乳杆菌GR-1介导MAPK信号通路抗大肠杆菌诱导奶牛子宫内膜上皮细胞炎性损伤  

Protective effect of MAPK signal pathway mediated by Lactobacillus rhamnosus GR-1 against inflammatory injury of bovine endometrial epithelial cells induced by E.coli

作　　者：刘佳玮 冯晓微 焦玉兰[1,3] 孙艳[1] 靳天雄 冯鸣鹊 刘蓓 刘明超 LIU Jiawei;FENG Xiaowei;JIAO Yulan;SUN Yan;JIN Tianxiong;FENG Mingque;LIU Bei;LIU Mingchao(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Veterinary Biological Technology Innovation Center of Hebei Province,Baoding 071001,China;Ringpu(Baoding)Biological Pharmaceutical Co.,Ltd,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]河北省兽医生物技术创新中心,河北保定071001 [3]瑞普(保定)生物药业有限公司,河北保定071001

出　　处：《河北农业大学学报》2022年第2期99-106,共8页Journal of Hebei Agricultural University

基　　金：国家自然科学基金(31902328);河北省自然科学基金(C2021204067);河北省省属高等学校基本科研业务费研究项目(KY2021008).

摘　　要：为探讨鼠李糖乳杆菌GR-1(Lactobacillus rhamnosus GR-1)防治奶牛子宫内膜炎的具体通路机制。本研究利用L. rhamnosus GR-1和大肠杆菌(Escherichia coli)处理原代培养的奶牛子宫内膜上皮细胞(Bovine endometrial epithelial cells,BEECs),设对照组、E. coli攻毒组、L. rhamnosus GR-1单独处理组、L. rhamnosus GR-1预处理再加入E. coli组,然后利用细胞凋亡试剂盒检测细胞凋亡情况;采用Western blot技术检测p38、JNK、ERK蛋白磷酸化水平;利用MAPK特异性信号通路抑制剂抑制目的信号转导;通过Realtime qPCR技术检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-8(IL-8)mRNA的表达水平。结果显示,与E. coli攻毒组相比,L. rhamnosus GR-1预处理组的奶牛子宫内膜上皮细胞的凋亡数量显著降低(P<0.05),且p-p38、p-JNK蛋白表达水平极显著降低(P<0.01),L.rhamnosusGR-1预处理组中TNF-α、IL-1β、IL-6和IL-8mRNA的表达水平与E.coli攻毒组相比显著下降(P<0.05),MAPK特异性信号通路抑制剂可显著抑制E. coli诱导的TNF-α、IL-1β、IL-6和IL-8 mRNA的表达(P<0.05),产生和L. rhamnosus GR-1类似的抗炎作用。本研究结果表明L. rhamnosus GR-1可通过介导MAPK信号通路调节E. coli诱导的BEECs的炎性损伤,为奶牛子宫内膜炎的防控提供有益参考。This study aimed to explore the specific pathway mechanism of Lactobacillus rhamnosus GR-1(L.rhamnosus GR-1)in the prevention and treatment of bovine endometritis.The primary cultured bovine endometrial epithelial cells(BEECs)were treated with L.rhamnosus GR-1 and Escherichia coli(E.coli)for 4 experiments including control group,E.coli challenge group,L.rhamnosus GR-1 single treatment group,L.rhamnosus GR-1 pretreatment followed by adding E.coli group.Then apoptosis was detected using apoptosis kit and protein phosphorylation of p38 and JNK were evaluated by Western blot.The specific inhibitor of MAPK signal pathway was used to inhibit the target signal transduction,and the mRNA expression levels of Tumor necrosis factor-α(TNF-α),Interleukin-1β(IL-1β),Interleukin-6(IL-6),and Interleukin-8(IL-8)were detected by Real-time qPCR.The results showed that compared with the group challenged with E.coli alone,the number of apoptosis of endometrial epithelial cells was lower(P<0.05)in the L.rhamnosus GR-1 pretreatment group,and the expression of p-p38,p-JNK and other proteins were significantly decreased(P<0.01)and the expression of TNF-α,IL-1β,IL-6,and IL-8 were significantly lower than that in the E.coli group(P<0.05).MAPK specific signal pathway inhibitor could significantly inhibit the expression of TNF-α,IL-1β,IL-6,and IL-8 induced by E.coli(P<0.05),resulting in an anti-inflammatory effect similar to that of L.rhamnosus GR-1.To sum up,L.rhamnosus GR-1 can regulate the inflammatory injury of BEECs induced by E.coli by inhibiting the MAPK signal pathway,which provided a useful reference for the prevention and control of endometritis in dairy cows.

关 键 词：奶牛子宫内膜上皮细胞 炎性反应 鼠李糖乳杆菌 大肠杆菌 丝裂原活化蛋白激酶 

分 类 号：S854.5[农业科学—临床兽医学]