短暂高糖通过组蛋白甲基化修饰诱导肾小球系膜细胞持续炎性反应  

Transient high glucose induces persistent inflammatory status in rat glomerular mesangial cell via histone methylation modification

作　　者：邓云蕾[1] 范秋灵[1] 汪旭[1] 曹旭[1] 徐莉[1] 刘佳[1] 赵雪[1] 王力宁[1] 

机构地区：[1]中国医科大学附属第一医院肾内科,沈阳110001

出　　处：《中华肾脏病杂志》2017年第3期213-218,共6页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81270808）;辽宁省自然科学基金（201602821）;辽宁省高等学校重大科技平台免疫皮肤病学重点实验室自主创新课题基金（201303）;沈阳市科技计划项目（F16-205-1-40）

摘　　要：目的探讨短暂高糖培养后再恢复正常糖环境的大鼠肾小球系膜细胞是否持续释放炎性因子，及该过程是否与组蛋白甲基化修饰有关。方法大鼠肾小球系膜细胞株（HBZY-1）分为高糖组（25．0mmol／L葡萄糖）、高渗对照组（19．5mmol／L甘露醇＋5．5mmol／L葡萄糖）和正常对照组（5．5mmol／L葡萄糖）进行24h培养；随后各组更换正常糖培养基（5．5mmol／L葡萄糖）培养0、24、48、72h。分别提取各组细胞的蛋白、上清液以及总RNA。采用Western印迹检测组蛋白H3四号位赖氨酸单甲基化（H3K4mel）表达水平，实时荧光定量PCR检测核因子κB（NF—κB）的p65亚基、组蛋白甲基化转移酶set7／9的mRNA表达，ELISA检测上清中单核细胞趋化蛋白1（MCP-1）、血管细胞黏附分子1（VCAM-1）的表达。结果（1）与正常对照组比较，高糖处理大鼠系膜细胞24h后set7／9mRNA表达和H3K4mel蛋白表达均上调（均P〉0．05）；恢复正常糖培养24、48、72h高糖组set7／9mRNA表达和H3K4mel蛋白表达逐渐降低（与0h高糖组比较，均P〈0．05）；恢复正常糖培养72h高糖组set7／9和H3K4mel的表达与正常对照组差异均无统计学意义（均P〉0．05）。（2）未恢复正常糖培养（0h）时高糖组大鼠系膜细胞NF-κBp65mRNA、MCP-1、VCAM-1的表达均高于正常对照组（均P〉0．05）；恢复正常糖培养24、48h和72h高糖组NF—κBp65mRNA、MCP-1、VCAM-1的表达均保持高表达，与0h高糖组比较差异均无统计学意义（均P〉0．05）。以上各指标高渗组与正常对照组比较差异均无统计学意义（均P〉0．05）。结论短暂高糖培养可诱导。肾小球系膜细胞组蛋白甲基化转移酶set7／9和组蛋白H3K4单甲基化的表达上调，并且在恢复正常糖环境后仍持续48h。同时短暂高糖培养促进肾小球系膜细胞NF—κBp65，炎性因子MCP-1、VCAM-1的表达持续高表达。提示高糖代谢记忆可能�Objective To investigate whether the effect of transient high glucose on inflammatory factors expression could be continuous in rat glomerular mesangial cell, and its relation with histone methylation modification. Methods Rat glomerular mesangial cells （HBZY- 1） were divided into three groups： the high glucose group （25.0 mmol/L glucose）, the hypertonic group （MA, 5.5 mmol/L glucose＋ 19.5 mmol/L mannitol） and the normal-glucose control group （5.5 mmol/L glucose）, which were cultured for 24 h respectively. All 3 groups were then changed with normalglucose medium to culture for 24 h, 48 h and 72 h. Their protein, mRNA and supernatant were harvested. The protein expressions of mono-methylation of H3 lysine g （H3K4mel） was measured by Western blotting, and the mRNA expressions of NF - κB subunit p65 and setT/9 were determined by real time - quantitative PCR. The expression of monocyte chemoattractant protein 1 （MCP- 1） and vascular cell adhesion molecule 1 （VCAM-1） were detected by enzyme-linked immunosorbent assay. Results （1） Compared with those in normal control group, the expressions of H3K4mel protein and setT/9 mRNA were first up-regulated in high glucose group, then gradually down- regulated in the following d8 h normal-glucose medium （as compared with those at 0 h, all P 〈 0.05）. At 72 h there was no statistic difference between high glucose group and normal control group （all P 〉 0.05）. （2） Compared with those in normal control group, the up-regulated p65 mRNA, VCAM-1 and MCP-1 sustained at least for 72 h in high glucose group. Conclusions Transient high glucose can induce persistent inflammatory factors expression in rat glomerular mesangial cells, which may via histone modification.

关 键 词：糖尿病肾病 甲基化 血管细胞黏附分子1 趋化因子CCL2 代谢记忆 

分 类 号：R692.9[医药卫生—泌尿科学] R587.2[医药卫生—外科学]