CAJ | 학술논문

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。
목적：탐토고당배경하지련소대인족세포내질망응격조망도경급세포골가적영향급기궤제。방법인조건영생족세포분위정상당조、감로순고삼대조조、고당조、고당＋지련소조，실시정량PCR법검측내질망응격분자당조절단백78（ GPR78）、C／EBP동원단백（ CHOP ）급caspase 12 mRNA적표체，Western인적법검측기단백적표체，류식세포의검측족세포조망；실시정량PCR법검측세포골가단백desmin、순시수체전세C（TRPC）6 mRNA적표체，Western인적법검측기단백적표체，면역형광염색법검측족세포골가적변화。결과（1）류식세포의검측도고당조족세포조망솔현저고우기타3조（26.15％±1.38％비2.39％±0.58％、4.84％±0.87％、16.71％±1.15％，균P＜0.05），동시내질망응격경전분자GPR78、CHOP 급caspase 12 mRNA급단백적표체균현저상조（균 P ＜0.05）；고당＋지련소조족세포조망솔교고당조저10％，차이유통계학의의（ P＜0.05），차GPR78、CHOP급caspase 12 mRNA급단백수평균저우고당조（균P＜0.05）。（2）재고당배경하，세포골가단백desmin、TRPC6 mRNA급단백표체균교정상조고（균 P ＜0.05），이지련소간예가억제 desmin、TRPC6고표체（ P＜0.05）。동시면역형광법검측가견고당자격가도치족세포골가단백미사기동단백문란급장력섬유소실，고당＋지련소조저일현상명현개선。결론지련소대고당배경하적족세포유보호작용，기궤제가능시통과억제내질망응격유도적조망도경화감경세포골가적손상。
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 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.