茶树油对脂多糖诱导奶牛小肠上皮细胞损伤的影响  

Effects of Tea Tree Oil on Lipopolysaccharide-Induced Injury in Bovine Intestinal Epithelial Cells

作　　者：陈明[1] 蔡淑先 杨红 张佳豪 赵国琦[3] CHEN Ming;CAI Shuxian;YANG Hong;ZHANG Jiahao;ZHAO Guoqi(Jiangsu Vocational College of Agriculture and Animal Husbandry,Taizhou 225300,China;Nongan County Center for Animal Disease Control and Prevention,Changchun 130200,China;College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]江苏农牧科技职业学院,泰州225300 [2]农安县动物疫病预防控制中心,长春130200 [3]扬州大学动物科学与技术学院,扬州225009

出　　处：《动物营养学报》2024年第6期3942-3951,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31972589);现代农业产业技术体系专项资金资助(CARS-36);江苏省高等学校基础科学(自然科学)研究面上项目(23KJD230002)。

摘　　要：本试验旨在研究茶树油(TTO)对脂多糖(LPS)诱导奶牛小肠上皮细胞(BIECs)损伤的影响。试验选取3头健康新生中国荷斯坦犊牛的空肠组织,分离并获得BIECs进行培养。通过使用不同浓度的LPS(0、1、2和4μg/mL)和不同浓度的TTO(0、0.00625%、0.01250%、0.02500%、0.05000%和0.10000%),建立细胞炎症模型。然后,对照组以不含TTO和LPS的培养基处理细胞;LPS组以1μg/mL的LPS处理细胞12 h;LPS+TTO组采用0.01250%和0.02500%的TTO预处理细胞12 h,经水洗后,再暴露于LPS处理12 h。结果表明:1)与对照组(未添加TTO)相比,添加0.00625%、0.01250%、0.02500%和0.05000%TTO对BIECs细胞活力无显著影响(P>0.05),添加TTO显著提高跨膜电阻(TEER)(P<0.05)。与对照组相比,添加0.01250%TTO显著提高BIECs中闭锁小带蛋白-1(ZO-1)、封闭蛋白(occludin)、闭合蛋白-1(claudin-1)和闭合蛋白-4(claudin-4)的mRNA相对表达量(P<0.05),显著降低肿瘤坏死因子-α(TNF-α)的mRNA相对表达量(P<0.05)。2)与LPS组相比,LPS+TTO组TEER以及ZO-1、occludin和claudin-1的mRNA相对表达量显著提高(P<0.05),TNF-α的mRNA相对表达量显著降低(P<0.05);LPS+TTO组ZO-1蛋白相对表达量显著提高(P<0.05)。综上可知,TTO可以通过上调紧密连接蛋白表达和下调炎性因子表达,缓解LPS诱导的犊牛小肠上皮细胞屏障功能障碍和炎症损伤。This study was conducted to investigate the effects of tea tree oil(TTO)on lipopolysaccharide(LPS)-induced injury in bovine intestinal epithelial cells(BIECs).Jejunum tissues of 3 healthy newborn Chinese Holstein calves were isolated to obtain BIECs for culture.Cell inflammation model was established by using different concentrations of LPS(0,1,2 and 4μg/mL)and TTO(0,0.00625%,0.01250%,0.02500%,0.05000%and 0.10000%).Then,the cells in the control group were treated with medium without TTO and LPS,those in LPS group were treated with 1μg/mL LPS for 12 h,and those in LPS+TTO group were treated with 0.01250%and 0.02500%TTO for 12 h,and then exposed to LPS for 12 h after water washing.The results showed as follows:1)compared with the control group(without TTO),the supplementation of 0.00625%,0.01250%,0.02500%and 0.05000%TTO had no significant effects on the cell viability of BIECs(P>0.05),and the supplementation of TTO significantly increased the transepithelial electrical resistance(TEER)(P<0.05).Compared with the control group,the supplementation of 0.01250%TTO significantly increased the mRNA relative expression levels of zonula occludens-1(ZO-1),occludin,claudin-1 and claudin-4 in BIECs(P<0.05),and significantly decreased the tumor necrosis factor-α(TNF-α)mRNA relative expression level(P<0.05).2)Compared with LPS group,the TEER and mRNA relative expression levels of ZO-1,occludin and claudin-1 in LPS+TTO group were significantly increased(P<0.05),and the TNF-αmRNA relative expression level was significantly decreased(P<0.05);the ZO-1 protein relative expression level in LPS+TTO group was significantly increased(P<0.05).In conclusion,TTO can alleviate LPS-induced barrier dysfunction and inflammatory injury in intestinal epithelial cells of calves by up-regulating the expression of tight junction protein and down-regulating the expression of inflammatory factor.

关 键 词：茶树油 奶牛小肠上皮细胞 炎症 紧密连接 

分 类 号：S823[农业科学—畜牧学]