高糖环境下趋化素与其受体ChemR23通过活化p38MAPK促进肾小球内皮细胞炎性因子IL-6、TNF-α的表达  

Chemerin/ChemR23 promotes high glucose-induced IL-6 and TNF-α expressions in glomerular endothelial cells via p38 MAPK

作　　者：张晓雪[1] 王璐瑶[1] 尚进[1] 宁丽娜[1] 赵继芳[1] 窦艳娜[1] 郭佳[1] 肖静[1] 赵占正[1] 

机构地区：[1]郑州大学第一附属医院肾脏病医院肾内科,450052

出　　处：《中华肾脏病杂志》2017年第7期524-530,共7页Chinese Journal of Nephrology

摘　　要：目的观察趋化素（Chemerin）及其受体ChemR23在高糖刺激肾小球内皮细胞（GEnC）中的作用及机制。方法使用高糖体外刺激小鼠GEnC，分为正常对照组、20．0mmol／L高糖组、40．0mmol／L高糖组及渗透压对照组，检测培养上清中白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）的浓度，及细胞内Chemerin、ChemR23、IL-6和TNF-α的蛋白和mRNA的表达。应用慢病毒转染干扰ChemR23表达，并分别给予Chemerin及高糖刺激细胞，检测IL-6及TNF-α上清中的浓度和胞内mRNA的表达，及高糖刺激下p38丝裂原激活的蛋白激酶（p38MAPK）磷酸化水平。在高糖刺激前加入p38MAPK特异性抑制剂，检测培养上清IL-6、TNF-α的浓度，及其胞内mRNA的表达。各因子在培养上清中的浓度采用ELISA检测，Western印迹检测胞内蛋白表达和磷酸化水平，实时定量PCR检测胞内mRNA的表达。结果与对照组比较，20．0mmol／L和40．0mmol／L高糖组IL-6、TNF—α的表达及Chemerin蛋白和mRNA表达均增加（均P〈0．05），而ChemR23表达均无明显变化（均P〉0．05），Chemerin刺激组IL-6、TNF—α上清表达和胞内mRNA表达均增加（均尸〈0．05）。应用慢病毒干扰ChemR23表达后，Chemerin或高糖诱导的IL-6及TNF-α过表达均受到抑制（均P〈0．05）。同时，高糖组p38MAPK的磷酸化水平高于对照组（P〈0．05），而干扰ChemR23表达后，高糖刺激的p38MAPK磷酸化水平降低（与高糖组比较P〈0．05）。加入p38MAPK抑制剂，高糖诱导的IL-6、TNF-α过表达均降低（与高糖组比较均P〈0．05）。结论高糖刺激可诱导GEnC合成Chemerin，并通过其受体ChemR23引起p38MAPK信号通路的活化，从而诱导炎性因子IL-6、TNF-α的表达，促进GEnC的炎性反应。Objective To observe the role and related mechanism of chemerin and its receptor ChemR23 in glomerular endothelial cells （GEnCs） stimulated by high glucose. Methods Mouse GEnCs were cultured and divided into control group, 20.0 mmoL/L high glucose group, 40.0 mmol/L high glucose group and mannitol control group. Then the expressions of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF- α） in cell culture supernatant as well as the expressions of intracellular protein and mRNA of chemerin, ChemR23, IL-6 and TNF- a were detected. Lentiviral transfection targeting ChemR23 was applied before high glucose- or Chemerin- stimulated, and expressions of supernatant and intracellular mRNA of IL-6 and TNF-α were measured. Meanwhile whether p38 mitogen-activated protein kinase （p38 MAPK） pathway was activated by high glucose was detected. The specific inhibitor of p38 MAPK was added prior to high glucose - stimulated, then supernatant and intracellular mRNA expressions of IL-6 and TNF-α was detected. The supernatant expressions of IL-6 and TNF-α were measured by ELISA. The intracellular protein expression and p38 MAPK phosphorylation activity were detected by Western blotting. The mRNA expression was detected by real time PCR. Results Compared with those in the control group, in high glucose groups the expressions of IL-6, TNF- α and chemerin were significantly increased （all P 〈 0.05）, however, the expressions of ChemR23 did not change （all P 〉 0.05）; the supernatant and mRNA expressions of IL-6 and TNF-α were also elevated in the chemerin group （all P 〈 0.05）. Lentivirus baring shRNA could efficiently suppress ChemR23 expression, and the Chemerin- or high glucose-induced expressions of IL- 6 and TNF- α were reduced （all P 〈 0.05）. Also it could significantly reduce the expression of phosphorylated-p38 MAPK （p-p38 MAPK） induced by high glucose （P 〈 0.05）, as high glucose group had higher p-p38 MAPK than control group （P 〈 0.05）. While the high glucose-eleva

关 键 词：炎症趋化因子类 P38丝裂原活化蛋白激酶类 炎症 糖尿病肾病 肾小球内皮细胞 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]