摘要
目的：探讨高糖环境下脂联素对人足细胞内质网应激凋亡途径及细胞骨架的影响及其机制。方法人条件永生足细胞分为正常糖组、甘露醇高渗对照组、高糖组、高糖＋脂联素组，实时定量PCR法检测内质网应激分子糖调节蛋白78（ GPR78）、C／EBP同源蛋白（ CHOP ）及caspase 12 mRNA的表达，Western印迹法检测其蛋白的表达，流式细胞仪检测足细胞凋亡；实时定量PCR法检测细胞骨架蛋白desmin、瞬时受体电势C（TRPC）6 mRNA的表达，Western印迹法检测其蛋白的表达，免疫荧光染色法检测足细胞骨架的变化。结果（1）流式细胞仪检测到高糖组足细胞凋亡率显著高于其他3组（26.15％±1.38％比2.39％±0.58％、4.84％±0.87％、16.71％±1.15％，均P＜0.05），同时内质网应激经典分子GPR78、CHOP 及caspase 12 mRNA及蛋白的表达均显著上调（均 P ＜0.05）；高糖＋脂联素组足细胞凋亡率较高糖组低10％，差异有统计学意义（ P＜0.05），且GPR78、CHOP及caspase 12 mRNA及蛋白水平均低于高糖组（均P＜0.05）。（2）在高糖环境下，细胞骨架蛋白desmin、TRPC6 mRNA及蛋白表达均较正常组高（均 P ＜0.05），而脂联素干预可抑制 desmin、TRPC6高表达（ P＜0.05）。同时免疫荧光法检测可见高糖刺激可导致足细胞骨架蛋白微丝肌动蛋白紊乱及张力纤维消失，高糖＋脂联素组这一现象明显改善。结论脂联素对高糖环境下的足细胞有保护作用，其机制可能是通过抑制内质网应激诱导的凋亡途径和减轻细胞骨架的损伤。
Objective To explore the effects of adiponectin （ APN ） on high glucose-induced endoplasmic reticulum stress （ ERS ）-mediated apoptosis and cystoskeleton.Methods The conditionally immortal human glomeroular podocytes were divided into normal glucose , mannitol, high glucose and high glucose with adiponectin groups.Flow cytometry was employed to assess cell apoptosis.Real-time polymerase chain reaction （ PCR ） and Western blot were used to detect the expressions of such ERS molecules as GRP78, CHOP and caspase 12 and desmin and TRPC6.Immunofluorescent staining was used to detect the changes in the skeleton of podocyte.Results （1） The apoptosis rate in high glucose group is significantly higher than the other groups （ 26.15% ±1.38% vs 2.39% ±0.58%, 4.84% ±0.87%, 16.71% ± 1.15%, all P〈0.05）.Compared with control group , the mRNA and protein expressions of GRP 78, CHOP and caspase 12 were all up-regulated significantly in high glucose group （ P〈0.05）.The high glucose with adiponectin group could reduce the podocyte apoptosis by 10% and down-regulated the mRNA and protein expressions of GRP78, CHOP and caspase 12 versus high glucose group （ all P〈0.05）.（2） High glucose-induced podocyte caused the up-regulated expressions of TRPC 6 and desmin （ all P 〈0.05 ） and it was inhibited by adiponectin （ all P 〈0.05 ）.Additionally , imunofluorescent assay of high glucose-induced podocyte cytoskeleton showed disorderly F-actin and absent tensile fiber.Adiponectin prevented the F-actin＆amp;nbsp;cytoskeleton disruption under high glucose.Conclusion Adiponectin plays a protective role in high glucose-induced podocyte through reducing endoplasmic reticulum stress-induced apoptosis and blunting the injury of cytoskeleton.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2014年第14期1092-1096,共5页
National Medical Journal of China
基金
国家自然科学基金（30971380）
山西省研究生优秀创新项目（20123059）
关键词
足细胞
葡萄糖
内质网
细胞凋亡
细胞骨架
Podocytes
Glucose
Endoplasmic reticulum
Apoptosis
Cytoskeleton
