维生素D受体对LPS急性肾损伤过程肾脏氧化应激和炎性信号的调控作用

发布时间：2019-07-09 06:29

【摘要】：目的:本课题在前期成功建立LPS急性肾损伤模型基础上,通过深入研究体内和体外条件下维生素D3对LPS诱发小鼠肾脏和人肾小管上皮细胞炎性信号和氧化应激基因表达的抑制效应,阐明维生素D受体对LPS急性肾损伤过程肾脏氧化应激和炎性信号的调控作用和机制。方法:取96只小鼠随机分到四组,每组24只。LPS处理组小鼠通过腹腔注射LPS,剂量为2.0 mg/kg;生理盐水组小鼠通过腹腔注射生理盐水;LPS+维生素D3组小鼠注射LPS前48小时、24小时和1小时分别经口灌胃给予维生素D3;维生素D3组小鼠注射生理盐水前48小时、24小时和1小时分别通过灌胃方法给予维生素D3。各组小鼠于LPS注射后1小时、6小时或24小时使用二氧化碳和颈部脱臼方法处死。取血清及双侧肾脏,分别检测肾功能、MDA、GSH含量、尿素氮及肌酐水平、氧化酶基因及抗氧化酶基因,检测肾脏炎性因子和介质的表达、VDR的表达以及二者之间的相互关系,检测各条炎性通路表达及其与VDR的相互关系。结果:维生素D3预处理保护LPS引起的小鼠肾功能损害,激活小鼠肾脏VDR信号并促进其向核内转移,减轻LPS引起的肾脏GSH耗竭和蛋白硝化,减少血清和肾脏NO产生、下调肾脏i NOS表达,有效抑制LPS引起的肾脏氧化酶基因上调和抗氧化酶基因下调,阻断LPS引起的肾小管上皮细胞凋亡,对LPS引起的肾功能损伤起到保护作用;维生素D3预处理抑制LPS引起的小鼠肾炎性因子和介质上调,激活小鼠肾脏VDR信号;维生素D3抑制LPS诱导肾小管上皮细胞炎性因子和介质与VDR激活有关,维生素D3以I-κB非依赖方式阻断LPS诱导肾小管上皮细胞NF-κB p65向细胞核转移,维生素D3促进肾小管上皮细胞VDR与NF-κB p65相互作用。结论:维生素D3抑制LPS急性肾损伤过程肾脏氧化应激及炎性信号,其抑制作用与肾脏VDR信号激活有关。
文内图片：
图片说明：VitD3预处理减轻LPS引起的急性肾损伤
[Abstract]:Aim: based on the successful establishment of LPS acute renal injury model in the early stage, the inhibitory effects of vitamin D _ 3 on inflammatory signal and oxidative stress gene expression in mouse kidney and human tubular epithelial cells induced by LPS in vivo and in vitro were studied, and the regulatory effect and mechanism of vitamin D receptor on oxidative stress and inflammatory signal in kidney during acute renal injury in LPS were clarified. Methods: 96 mice were randomly divided into four groups with 24 mice in each group. The mice in the LPS, treatment group were injected with normal saline through intraabdominal injection, and the mice in the LPS vitamin D 3 group were given vitamin D 3 by oral administration 48 hours, 24 hours and 1 hour before injection of LPS, and the mice in the LPS vitamin D 3 group were given vitamin D 3 by oral administration 48 hours, 24 hours and 1 hour, respectively. Vitamin D 3 mice in vitamin D 3 group were given vitamin D 3 48 hours, 24 hours and 1 hour before injection of normal saline. The mice in each group were killed by carbon dioxide and cervical dislocations 1 hour, 6 hours or 24 hours after LPS injection. Serum and bilateral kidneys were taken to detect renal function, MDA,GSH content, urea nitrogen and creatinine levels, enzyme gene and antioxidant enzyme gene, the expression of inflammatory factors and mediators, the expression of VDR and the relationship between them, and the expression of each inflammatory pathway and its relationship with VDR. Results: vitamin D 3 pretreatment protected renal function damage induced by LPS, activated mouse kidney VDR signal and promoted its transfer to nucleus, alleviated renal GSH depletion and protein nitrification induced by LPS, reduced NO production in serum and kidney, down-regulated renal I NOS expression, effectively inhibited LPS induced up-regulation of renal enzyme gene and down-regulation of antioxidant enzyme gene, and blocked apoptosis of renal tubular epithelial cells induced by LPS. It has protective effect on renal function injury induced by LPS. Vitamin D 3 pretreatment inhibited the upregulation of inflammatory factors and mediators induced by LPS and activated VDR signal in mouse kidney. Vitamin D 3 inhibited LPS induced renal tubular epithelial cell inflammatory factors and mediators related to VDR activation. Vitamin D 3 blocked the nuclear transfer of LPS induced renal tubular epithelial cells NF- 魏 B p65 in an I-kappa B independent manner. Vitamin D 3 promoted the interaction between VDR and NF- 魏 B p65 in renal tubular epithelial cells. Conclusion: vitamin D 3 can inhibit renal oxidative stress and inflammatory signal during acute renal injury in LPS, and its inhibitory effect is related to the activation of VDR signal in kidney.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692.5
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本文编号：2511926