绿原酸对奶牛乳腺上皮细胞的保护作用及对中性粒细胞功能的影响

发布时间：2018-04-05 02:21

  本文选题：绿原酸　切入点：金黄色葡萄球菌　出处：《扬州大学》2017年硕士论文

【摘要】：本文主要研究了绿原酸对金黄色葡萄球菌感染的乳腺上皮细胞的保护作用及绿原酸对奶牛中性粒细胞功能的影响。试验一、绿原酸对金黄色葡萄球菌与奶牛乳腺上皮细胞相互作用的影响研究0、10、20、30 μg/mL绿原酸对金黄色葡萄球菌增殖的抑制作用。结果显示培养0、3、6 h时绿原酸对金黄色葡萄球菌的增殖没有显著影响,培养9、12 h时30 μg/mL绿原酸组对金黄色葡萄球菌增殖具有显著的抑制作用(P0.05)。培养24h时10 μg/mL绿原酸组对金黄色葡萄球菌增殖具有显著的抑制作用(P0.05),20、30μg/mL绿原酸组对金黄色葡萄球菌增殖具有极显著的抑制作用(P0.01)。20μg/mL绿原酸可以对已形成的细菌生物膜起到破坏作用(P0.05),其他组无显著影响。10、20、30μg/mL绿原酸对乳腺上皮细胞活性无负面影响,20 μg/mL绿原酸组可以显著提高金黄色葡萄球菌感染的乳腺上皮细胞的存活率(P0.05),其他组无显著影响。10、20、30 μg/mL绿原酸对金黄色葡萄球菌粘附侵袭乳腺上皮细胞没有显著影响,但30 μg/mL绿原酸组可以显著降低金黄色葡萄球菌对乳腺上皮细胞的侵袭率(P0.05)。30 μg/mL绿原酸组可以显著降低金黄色葡萄球菌诱导的乳腺上皮细胞ROS的产生(P0.05),其他组无显著影响。试验二、绿原酸对金黄色葡萄球菌诱导的奶牛乳腺上皮细胞炎症反应的影响为探究绿原酸对乳腺上皮细胞炎症相关因子表达的影响,利用金黄色葡萄球菌感染乳腺上皮细胞后加入绿原酸进行培养,结果显示30 μg/mL绿原酸可以极显著降低IL-1β、IL-6、IL-8、TLR-2mRNA 的表达(P0.01),对 TNF-α、MyD88mRNA 的表达无显著影响。30 μg/mL绿原酸组可以显著降低MAPK信号通路p38、JNK蛋白的磷酸化水平,对ERK亚族的磷酸化没有影响。试验三、绿原酸对中性粒细胞功能及炎症相关基因表达的影响为探究绿原酸对中性粒细胞功能的影响,对中性粒细胞的趋化、吞噬、杀菌功能进行评价。分别选用0、10、20、30 μg/mL绿原酸与中性粒细胞共培养,结果显示10、20、30 μg/mL绿原酸对中性粒细胞活性没有影响,10、20、30 μg/mL绿原酸对中性粒细胞的趋化能力没有影响,20μg/mL绿原酸组可以显著提高中性粒细胞的吞噬能力(P0.05),30μg/mL组可以极显著提高吞噬能力(P0.01)。添加20μg/mL绿原酸组中性粒细胞的杀菌能力显著提高(P0.05),其他组无显著影响。为探究绿原酸对中性粒细胞炎症相关因子表达的影响,对趋化实验下室的中性粒细胞基因表达进行分析,结果显示,30μg/mL绿原酸组可以降低中性粒细胞IL-1β、TLR-2mRNA的表达(P0.05),但对TGF-β、IL-8 mRNA的表达无显著影响。
[Abstract]:The protective effect of Lv Yuan acid on breast epithelial cells infected with Staphylococcus aureus and the effect of Lv Yuan acid on the function of neutrophils in dairy cattle were studied.Experiment 1. The effect of Lv Yuan acid on the interaction between Staphylococcus aureus and mammary epithelial cells of dairy cows was studied. The inhibitory effect of 0 ~ (10) ~ (20) ~ (20) 渭 g/mL Lv Yuan acid on the proliferation of Staphylococcus aureus was studied.The results showed that Lv Yuan acid had no significant effect on the proliferation of Staphylococcus aureus at 0 ~ 3h for 6 h, and the 30 渭 g/mL Lv Yuan acid group had a significant inhibitory effect on the proliferation of Staphylococcus aureus at 9 ~ 12 h.Inhibitory effect of 10 渭 g/mL Lv Yuan acid group on Staphylococcus aureus proliferation at 24 h. The inhibitory effect of 10 渭 g/mL Lv Yuan acid group on Staphylococcus aureus proliferation was significant. P0.01- .20 渭 g/mL Lv Yuan acid could inhibit the proliferation of Staphylococcus aureus biofilm.There was no significant effect on the activity of mammary epithelial cells in the other group. There was no negative effect on the activity of mammary epithelial cells in the other groups. The survival rate of mammary epithelial cells infected with Staphylococcus aureus was significantly increased in the 20 渭 g/mL Lv Yuan group, and no significant difference was found in the other groups, while in the other groups, the survival rate of breast epithelial cells infected with Staphylococcus aureus was significantly increased.The effect of #number0# 渭 g/mL Lv Yuan on Staphylococcus aureus adhesion and invasion of breast epithelial cells was not significant.However, 30 渭 g/mL Lv Yuan acid group could significantly reduce the invasion rate of Staphylococcus aureus on breast epithelial cells. 30 渭 g/mL Lv Yuan acid group could significantly reduce the ROS production induced by Staphylococcus aureus, but the other groups had no significant effect.In experiment 2, the effect of Lv Yuan acid on the inflammatory response of bovine mammary epithelial cells induced by Staphylococcus aureus was to explore the effect of Lv Yuan acid on the expression of inflammatory related factors in mammary epithelial cells.Staphylococcus aureus was infected with mammary epithelial cells and cultured with Lv Yuan acid.缁撴灉鏄剧ず30 渭g/mL缁垮師閰稿彲浠ユ瀬鏄捐憲闄嶄綆IL-1尾,IL-6,IL-8,TLR-2mRNA 鐨勮〃杈，

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