LPS对奶牛乳腺上皮细胞促炎症因子基因表达的影响  

Effect of lipopolysaccharide on proinflammatory cytokines expression in bovine mammary epithelial cells

作　　者：宋金婷 张金友 庄雨龙[2] 李伟[2] 宋伟红[2] 李长志[2] SONG Jinting;ZHANG Jinyou;ZHUANG Yulong;LI Wei;SONG Weihong;LI Changzhi(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319,China;Instituteof Animal Husbandry and Veterinary,Heilongjiang Academy of Land Reclamation Science,Harbin 150038,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江省农垦科学院畜牧兽医研究所,哈尔滨150038

出　　处：《黑龙江动物繁殖》2022年第2期7-12,共6页Heilongjiang journal of animal reproduction

基　　金：黑龙江省自然科学基金联合引导项目“AMPK对牛乳腺上皮细胞先天性免疫因子表达影响研究”(LH2019C082);黑龙江省农垦总局重点科研计划项目“奶牛乳房炎对乳腺组织基因差异性表达影响及其与能量水平关系的研究”(HKKYZD190304);黑龙江八一农垦大学学成、引进人才科研启动计划“奶牛乳腺组织基因差异性表达及其AMPK水平研究”(XYB202017)。

摘　　要：为了探讨脂多糖(lipopolysaccharide,LPS)对奶牛乳腺上皮细胞中促炎症因子基因表达的影响,试验采用酶消化法将奶牛乳腺上皮细胞分离纯化后分成两组,分别在含0.5 mg/mL葡萄糖和0.5 mg/mL葡萄糖+1μg/mL LPS的完全培养液中培养24 h,采用荧光定量PCR技术检测细胞中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、转化生长因子-β1(transforming growth factor-beta 1,TGF-β1)、诱导型一氧化氮合酶(induced nitric oxide synthase,iNOS)、白细胞介素-6(interleukin-6,IL-6)、乳铁蛋白(lactoferrin,LF)和白细胞介素-8(Interleukin-8,IL-8)基因mRNA表达量。结果表明:TGF-β1基因的mRNA相对表达量在两组之间没有显著差异(P>0.05);而TNF-α、iNOS、IL-6、LF和IL-8基因的mRNA相对表达量在LPS刺激后均提高,其中IL-6和LF基因的mRNA相对表达量显著提高(P<0.05),TNF-α、iNOS和IL-8基因的mRNA相对表达量极显著提高(P<0.01)。说明奶牛乳腺上皮细胞受到感染后具有迅速增加诸多先天性免疫因子的能力,以便在乳腺感染的初期可以快速启动免疫识别和信号传递等免疫机制。The purpose of this study was to investigate the effects of lipopolysaccharide(LPS)on mRNA expressions of tumor necrosis factor-α(TNF-α),transforming growth factor beta 1(TGF-β1),inducible nitric oxide synthase(iNOS)and Interleukin-6(IL-6),lactoferrin(LF),Interleukin-8(IL-8)in bovine mammary epithelial cells.The mammary epithelial cells were separated and purified by enzyme digestion,and then divided into two groups.They were cultured for 24 hours in complete medium containing only 0.5 mg/mL glucose and 0.5 mg/mL glucose+1μg/mL LPS,respectively.The expressions of related genes in cells were detected by fluorescence quantitative PCR.The results showed that there was no significant difference in the relative mRNA expression of TGF-β1 gene between the two groups(P>0.05).While the expressions of TNF-α,iNOS,IL-6,LF and IL-8 mRNA were increased after LPS stimulation,in which the expressions of IL-6 and LF mRNA were significantly increased(P<0.05),and the expressions of TNF-α,iNOS and IL-8 mRNA were extremly increased(P<0.01).This study suggested that the mammary epithelial cells of dairy cows had the ability to rapidly increase many innate immune factors after being infected,so as to quickly start the immune mechanism such as immune recognition and signal transmission in the early stage of breast infection.

关 键 词：奶牛 乳腺上皮细胞 脂多糖 炎症 免疫 

分 类 号：Q786[生物学—分子生物学]