摘要
目的通过观察高糖培养下人脐静脉内皮细胞(HUVEC)的CD26表达变化、甘露聚糖结合凝集素(MBL)补体途径的激活及炎症因子(IL-6和NF-kB)的表达,探讨CD26在糖尿病肾病(DN)慢性炎症发病机制中的作用。方法体外培养HUVEC,分别设置对照组、高渗组、高糖组,采用免疫组化法及Western Blot检测细胞表面CD26、MBL、C3和膜攻击复合物(MAC)蛋白,采用实时荧光定量PCR(qRT-PCR)检测细胞中CD26、IL-6和NF-kB的m RNA。结果与对照组及高渗组相比,高糖组CD26、MBL、C3、MAC的蛋白表达量明显升高(P<0.05),且高糖组CD26、IL-6、NF-kB mRNA的相对表达量较其余两组亦明显升高(P<0.05);而高渗组与对照组相比,各指标蛋白及mRNA表达量均无统计学意义(P>0.05)。结论高糖作用下的HUVEC CD26蛋白表达量是增高的,其作用机制可能是通过激活MBL介导的甘露糖结合凝集素补体途径,使炎症因子IL-6、NF-kB表达增加,从而引起细胞的炎症反应。
Objective The study aims to observe the effects of high sugar on the expression of CD26,the activation of mannan binding lectin( MBL) complement pathway,and the expressions of inflammatory factors( IL-6 and NF-kB) in cultured human umbilical vein endothelial cells( HUVEC),and explore the role of CD26 in the pathogenesis of chronic inflammation that contribute to the development of diabetic nephropathy. Methods HUVEC were randomly divided into 3 groups as follow: control group with normal concentration of glucose,hypertonic group,and high glucose group. The depositions of CD26,MBL,C3 and MAC were respectively detected by immunohistochemical method and Western Blot,and the m RNA expressions of CD26,IL-6 and NF-kB were measured by qRT-PCR. Results Compared with the control group and the hypertonic group,the protein levels of CD26,MBL,C3 and MAC in the high glucose group were obviously increased( P < 0. 05),and the mRNA expressions of CD26,IL-6,NF-kB were also apparently increased( P < 0. 05). But there were no significant differences in the gene and protein expressions between the control group and the hypertonic group. Conclusion High concentration of glucose can increase the expression of CD26,MBL,C3,MAC,IL-6 and NF-kB in the cultured HUVEC. Increased expression of CD26 may promote the downstream inflammatory reaction through indirectly activating the MBL pathway and directly activating inflammatory factors( IL-6 and NF-kB).
引文
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