α-倒捻子素对奶牛乳房炎源性金黄色葡萄球菌诱导奶牛乳腺上皮细胞促炎反应的抑制作用  

α-Mangostin Inhibition of Proinflammatory Reaction in Bovine Mammary Epithelial Cells Induced by Staphylococcus aureus from Bovine Mastitis

作　　者：傅业全 侯晓林[2] 崔德凤[2] 胡格[2] 王建舫[2] 刘洋[1] 韩博[1] FU Ye-quan;HOU Xiao-lin;CUI De-feng;HU Ge;WANG Jian-fang;LIU Yang;HAN Bo(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]中国农业大学动物医学院,北京海淀100193 [2]北京农学院动物科学技术学院,北京昌平102206

出　　处：《中国兽医杂志》2021年第7期7-12,16,共7页Chinese Journal of Veterinary Medicine

基　　金：国家“十三五”重点研发计划(2016YFD0501307);宁夏回族自治区重点研发计划项目(2019BBF02027)。

摘　　要：本试验旨在探索α-倒捻子素对奶牛乳房炎源性金黄色葡萄球菌诱发感染奶牛乳腺上皮细胞(bMECs)促炎反应的抑制作用。用微量稀释法测定α-倒捻子素对金黄色葡萄球菌的最小抑菌浓度(MIC);CCK-8法测定α-倒捻子素的细胞毒性作用;RT-PCR和ELISA法分别测定细胞的TNF-α、IL-1β和IL-6 mRNA和蛋白表达水平;Western Blot法测定NF-κB p-p65蛋白的表达量。结果显示:α-倒捻子素对金黄色葡萄球菌的MIC为1.6μg/mL;α-倒捻子素在0~3.2μg/mL范围内对bMECs增殖没有明显的影响(P>0.05);添加不同终浓度的α-倒捻子素(0.4、0.8μg/mL和1.6μg/mL)不仅均能抑制金黄色葡萄球菌入侵bMECs(P<0.01),还能显著抑制金黄色葡萄球菌感染bMECs引起的TNF-α、IL-1β和IL-6 mRNA转录水平和蛋白水平的升高(P<0.01);金黄色葡萄球菌感染bMECs后可显著上调NF-κB p65蛋白磷酸化水平(P<0.01),添加0.4μg/mL和1.6μg/mL终浓度的α-倒捻子素可显著降低p65蛋白磷酸化水平(P<0.01),但是添加0.8μg/mL终浓度的α-倒捻子素也能降低p65蛋白的磷酸化水平,但无统计学意义(P>0.05)。结果表明α-倒捻子素对金黄色葡萄球菌有较强的抗菌作用,能够显著抑制金黄色葡萄球菌对bMECs的侵袭,有效缓解金黄色葡萄球菌感染bMECs引起的TNF-α、IL-1β和IL-6表达水平的上调,其抗炎的分子机理可能与NF-κB信号通路相关。This study investigated the ability ofα-mangostin in inhibit pro-inflammatory reaction in bovine mammary epithelial cells(bMECs)induced by Staphylococcus aureus from bovine mastitis.The minimal inhibitory concentration(MIC)was measured by broth microdilution method;the cytotoxic effect ofα-mangostin was tested by CCK-8;the mRNA and protein expression levels of TNF-α,IL-1βand IL-6 were determined by RT-PCR and ELISA,respectively.The expression level of NF-κB p-p65 protein was detected by Western Blot.The results showed that the MIC ofα-mangostin was 1.6μg/mL;α-mangostin in the range of 0-3.2μg/mL had no significant effect on the proliferation of bMECs(P>0.05);different doses ofα-mangostin(0.4,0.8μg/mL and 1.6μg/mL)not only inhibited the invasion of S.aureus into bMECs(P<0.01),but also significantly inhibited the expression levels of TNF-α,IL-1βand IL-6 mRNA and protein in bMECs induced by S.aureus(P<0.01).In addition,α-mangostin(at 0.4 and 1.6μg/mL)significantly decreased the expression level of NF-κB p-p65 protein in bMECs induced by S.aureus compared to noα-mangostin control(P<0.01),althoughα-mangostin(at 0.8μg/mL)did not significantly alter the NF-κB p-p65 expression level in bMECs induced by S.aureus compared to noα-mangostin control(P>0.05).By significantly inhibiting the growth and invasion of S.aureus originated from bovine mastitis,α-mangostin plays an anti-inflammatory role through suppression of TNF-α,IL-1βand IL-6 expression in bMECs induced by S.aureus,with the mechanism possibly related to NF-κB signal pathway.

关 键 词：α-倒捻子素 奶牛乳房炎 金黄色葡萄球菌 奶牛乳腺上皮细胞 炎性因子 NF-ΚB信号通路 

分 类 号：S823[农业科学—畜牧学]