发酵乳杆菌对LTA诱导的牛子宫内膜细胞炎性损伤干预作用的研究  

Intervention Mechanisms of Lactobacillus fermentum on LTA-induced Inflammatory Injury of Endometrial Cells in Bos Taurus

作　　者：杨淑华[1] 王佳美 李鹏[1] 龙淼[1] 李林[1] 张诗萌 杨靓 李康 何剑斌[1] YANG Shu-hua;WANG Jia-mei;LI Peng;LONG Miao;LI lin;ZHANG Shi-meng;YANG Liang;LI Kang;HE Jian-bin(College of Animal Husbandry and Veterinary Medicine,Shenyang Agricultural University,Shenyang 110161,China)

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110161

出　　处：《沈阳农业大学学报》2019年第1期1-9,共9页Journal of Shenyang Agricultural University

基　　金：沈阳市重大共性关键技术创新专项项目(17-165-3-00);沈阳市重点科技研发技术项目(18-004-3-45)

摘　　要：旨在研究发酵乳杆菌能否通过抑制NF-kB信号通路的激活,调控下游炎性细胞因子,进而对脂磷壁酸(LTA)所诱导的牛子宫内膜细胞(BEND)炎性损伤发挥保护作用,并通过体外试验对其作用机制进一步研究,为解决牛子宫内膜炎的治疗难题提供方向。以牛子宫内膜细胞为研究对象,LTA和发酵乳杆菌分别作为毒药与解药来建立体外细胞试验模型,测定细胞因子TNF-α、IL-1β、IL-6、IL-8的分泌量和炎性相关基因TNF-α、IL-1β、IL-6、IL-8、TLR2、MyD88、NF-kB p65 mRNA的表达量。结果表明:LTA组、LTA+Lf组、Lf组炎性细胞因子TNF-α、IL-1β、IL-6、IL-8的蛋白表达水平均高于对照组。LTA组与对照组比较差异极显著(p<0.01);Lf组与对照组比较,TNF-α、IL-6细胞因子差异不显著(p>0.05),IL-1β、IL-8细胞因子差异显著(p<0.05);LTA+Lf组与LTA组比较,TNF-α、IL-1β、IL-6、IL-8的蛋白表达量显著降低(p<0.05)。LTA组、LTA+Lf组、Lf组的TNF-α、IL-1β、IL-6、IL-8、TLR2、MyD88、NF-kB p65基因mRNA的表达量均高于对照组。LTA组与对照组相比,7种基因m RNA的表达量极显著增加(p<0.01);Lf组与对照组相比,TNF-α、IL-6基因mRNA的表达量增加不显著(p>0.05),IL-8基因mRNA的表达量增加显著(p<0.05),IL-1β、TLR2、MyD88、NF-kB p65基因mRNA的表达量极显著增加(p<0.01);LTA+Lf组与LTA组相比,7种基因mRNA的表达量均极显著降低(p<0.01)。发酵乳杆菌能够抑制NF-kB信号传导通路,调控炎性细胞因子的表达与释放,抑制炎性反应的发生,缓解LTA的细胞毒性,对LTA诱导的牛子宫内膜细胞炎性损伤起干预作用。This study is aim to investigate whether to play a protective effect of Lactobacillus fermentum on lipoteichoic acid(LTA)isolated from Staphylococcus aureus-induced inflammatory injury of endometrial cells in Bos taurus by inhibiting the activation of NF-kB signaling pathway and regulating the expression of downstream inflammatory cytokines.The mechanism of action through in vitro testing provided a reference for prevention and treatment of staphylococcus aureus-induced bovine endometritis.Establishing cell experiment model in vitro by taking endometrial cells in Bos taurus as the research object,LTA and Lactobacillus fermentum as poisons and antidotes and testing the secretion of inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-8)and the expression of inflammatory related genes mRNA(TNF-α,IL-1β,IL-6,IL-8,TLR2,MyD88,NF-kB p65).The results showed that the protein expression levels of inflammatory cytokines TNF-α,IL-1β,IL-6 and IL-8 in LTA group,LTA+Lf group and Lf group were higher than those in control group.There was a significant difference between the LTA group and the control group(p<0.01).Compared with the control group,the TNF-αand IL-6 cytokines were not significantly different(p >0.05),and the IL-1βand IL-8 cytokines were different significantly(p<0.05).The protein expression levels of TNF-α,IL-1β,IL-6 and IL-8 were significantly lower in the LTA+Lf group than those in the LTA group.The expression levels of TNF-α,IL-1β,IL-6,IL-8,TLR2,MyD88 and NF-kB p65 mRNA in LTA group,LTA+Lf group and Lf group were higher than those in control group.LTA group compared with the control group,the mRNA expression level of the seven kinds increased significantly(p<0.01).The mRNA expression of IL-6 and TNF-αin Lf group does not increase significantly(p >0.05),compared with the control group.The expression of IL-8 gene mRNA increased significantly(p<0.05).The expression of IL-1β,TLR2,MyD88,NF-kB p65 gene mRNA increased significantly(p<0.01).The mRNA expression levels of the seven genes in LTA+Lf group were significan

关 键 词：LTA 发酵乳杆菌 牛子宫内膜细胞 NF-kB信号传导通路 炎性损伤 

分 类 号：S858.23[农业科学—临床兽医学]