沙格列汀抑制NF-κB信号通路下调LPS诱导的人肾小管上皮HK-2细胞MCP-1的表达*

李清福1夏缨2李晓艳1何彦蓉1张维3李露4

(成都军区机关医院 1.内分泌科; 2.第二门诊部, 四川 成都 610000;3.成都医学院第一附属医院， 四川 成都 610500;

4.四川大学华西医院，四川 成都 610041)

【摘要】目的探讨二肽基肽酶4(DPP-4)竞争性抑制剂沙格列汀(saxagliptin)对脂多糖(LPS)诱导状态下人肾小管上皮HK-2细胞MCP-1表达的调节作用及相关的分子机制。方法将人肾小管上皮细胞HK-2细胞分为3组，分别为对照组(Control组)、LPS+沙格列汀 (LPS+SAX组)和LPS组。在干预12小时后使用RT-PCR法和western blot法对HK-2细胞MCP-1 mRNA和蛋白的表达水平进行检测；并使用western blot法对HK-2细胞NF-κB p65活化水平(phospho-NF-κB p65/total NF-κB p65比值)进行分析。结果与Control组相比较，在LPS干预12小时后 HK-2细胞MCP-1 mRNA和蛋白的表达水平以及phospho-NF-κB p65/total NF-κB p65比值均明显升高，差异均有统计学意义(P<0.05)；但LPS组较LPS+SAX组MCP-1 mRNA和蛋白的表达的升高以及phospho-NF-κB p65/total NF-κB p65比值的增加更加显著，差异均有统计学意义(P<0.05)。结论DPP-4抑制剂沙格列汀可以通过抑制NF-κB的磷酸化下调LPS诱导的人肾小管上皮HK-2细胞MCP-1合成和释放并对其产生保护作用。

【关键词】沙格列汀； HK-2细胞； 单核细胞趋化蛋白-1 (MCP-1)； 核转录因子-κB (NF-κB)

【中图分类号】R 446

【文献标志码】A

doi：10.3969/j.issn.1672-3511.2015.10.005

Abstract【】ObjectiveTo explore whether saxagliptin could down-regulate LPS-induced MCP-1 expression in human proximal tubular epithelial cells, and its potential molecular mechanism. MethodsHuman proximal tubular epithelial cells (HK-2 cells) were divided into 3 groups, Control group, LPS+saxagliptin group (LPS+SAX group) and LPS group. RT-PCR was used to measure the mRNA expression of MCP-1. The protein expression of MCP-1 and the phosphorylation level of NF-κB p65 were analyzed by western blot. ResultsAfter 12 hours LPS stimulation, the mRNA and protein expression of MCP-1 were remarkably up-regulated, and the ratio of phospho-NF-κB p65/total NF-κB p65 were sharply raised. Then, LPS-induced the up-regulation of MCP-1 and the raised ratio of phospho-NF-κB p65/total NF-κB p65 were all noticeably inhibited by saxagliptin in HK-2 cells. ConclusionSaxagliptin down-regulates LPS-induced MCP-1 expression possibly via inhibition of NF-κB in human proximal tubular epithelial cells (HK-2 cells).

基金项目：四川省教育厅科研课题(11ZB172)。

通讯作者：李清福，E-mail:liqingfucdjq2010@163.com

收稿日期：( 2014-10-10； 编辑： 陈舟贵)

Saxagliptin down-regulates LPS-induced MCP-1 expression via inactivation of NF-κB in human proximal tubular epithelial cellsHE Rong1, LI Qingfu1, XIA Ying2,etal

(1.DepartmentofEndocrinology,TheAffiliatedHospitalofChengduMilitaryRegion,Chengdu610000,China;

2.DepartmentofTheSecondOutpatient,TheAffiliatedHospitalofChengduMilitaryRegion,Chengdu610500,China;

3.TheFirstAffiliatedHospitalofChengduMedicalCollage,Chengdu610000,China;

4.WestChinaHospital,SichuanUniversity,Chengdu610041,China)

【Key words】Saxagliptin; HK-2 cells; Monocyte chemotactic protein-1; NF-κB

胰高糖素样肽-1 (glucagon-like peptide-1, GLP-1)属于肠肽类激素(incretin)，主要由肠道内L细胞合成和分泌，具有包括调节胰岛素分泌、糖代谢和脂代谢等多种生理效应[1～4]。进一步的研究发现GLP-1还具有调节能量代谢以外的包括抑制炎症反应、抑制氧化应激、抗器官纤维化以及调节细胞凋亡等多种生物学效应[2～9]。同时发现二肽基肽酶4 (dipeptidyl peptidase, DPP-4)是GLP-1代谢最主要的关键酶，抑制DPP-4可以有效的减少和延迟GLP-1的分解和代谢[2～9]。DPP-4竞争性抑制剂和GLP-1拟似物已广泛应用于Ⅱ型糖尿病的临床治疗[10]。本研究主要观察DPP-4竞争性抑制剂沙格列汀(saxagliptin)对脂多糖(LPS)诱导损伤状态下的人肾小管上皮HK-2细胞的保护作用及相关的分子机制。……