CAJ | 학술논문

目的：探讨过度内质网应激在慢性环孢素A（ CsA）肾毒性细胞凋亡中的作用机制。方法：将Spra-gue-Dawley大鼠分为正常对照组和慢性CsA肾毒性组，分别给予皮下注射橄榄油（1 mg · kg-1· d-1）和CsA （15 mg· kg-1· d-1）1周或4周后处死大鼠。三色染色和TUNEL染色观察肾小管间质纤维化和细胞凋亡；免疫组织化学染色和免疫印迹检测免疫球蛋白结合蛋白（ BiP）、磷酸化的真核细胞翻译起始因子2α（ eIF2α）、生长阻滞及DNA损伤诱导蛋白153（GADD153）、caspase-12和caspase-3蛋白表达。结果： CsA注射1周观察不到肾小管间质纤维化和TUNEL阳性细胞，而给予CsA注射4周大鼠表现为明显的肾小管间质纤维化[（38．9±3．3）％vs （0．0±0．0）％， P＜0．01]和大量TUNEL阳性细胞[（89±9）％ vs （7±2）％， P＜0．01]。与对照组比较，CsA组BiP和caspase-12蛋白的表达于1周达到高峰，4周后降至正常水平；反之，eIF2α和caspase-3蛋白表达呈时间依赖性增加。结论：在慢性CsA肾毒性中，过度的内质网应激耗尽分子伴侣，激活凋亡途径，从而导致肾小管上皮细胞凋亡和肾小管间质损伤。
목적：탐토과도내질망응격재만성배포소A（ CsA）신독성세포조망중적작용궤제。방법：장Spra-gue-Dawley대서분위정상대조조화만성CsA신독성조，분별급여피하주사감람유（1 mg · kg-1· d-1）화CsA （15 mg· kg-1· d-1）1주혹4주후처사대서。삼색염색화TUNEL염색관찰신소관간질섬유화화세포조망；면역조직화학염색화면역인적검측면역구단백결합단백（ BiP）、린산화적진핵세포번역기시인자2α（ eIF2α）、생장조체급DNA손상유도단백153（GADD153）、caspase-12화caspase-3단백표체。결과： CsA주사1주관찰불도신소관간질섬유화화TUNEL양성세포，이급여CsA주사4주대서표현위명현적신소관간질섬유화[（38．9±3．3）％vs （0．0±0．0）％， P＜0．01]화대량TUNEL양성세포[（89±9）％ vs （7±2）％， P＜0．01]。여대조조비교，CsA조BiP화caspase-12단백적표체우1주체도고봉，4주후강지정상수평；반지，eIF2α화caspase-3단백표체정시간의뢰성증가。결론：재만성CsA신독성중，과도적내질망응격모진분자반려，격활조망도경，종이도치신소관상피세포조망화신소관간질손상。
AIM:To investigate the impact of excessive endoplasmic reticulum stress on apoptotic cell death in a rat model of chronic cyclosporine A ( CsA ) nephrotoxicity .METHODS: Male Sprague-Dawley rats on a low-salt diet were subcutaneously injected with vehicle (olive oil, 1 mL· kg-1· d-1) or CsA (15 mg/kg) daily for 1 or 4 weeks.Tu-bulointerstitial fibrosis and apoptotic cell death were estimated by trichrome staining and TUNEL staining .In addition , im-munohistochemistry and immunoblotting were used to evaluate the expression of immunoglobulin -binding protein ( BiP) , eu-karyotic initiation factor 2α(eIF2α), growth arrest and DNA damage-inducible protein 153 (GADD153), caspase-12 and caspase-3.RESULTS:The rats treated with CsA for 1 week did not develop tubulointerstitial fibrosis and TUNEL-positive cells, whereas 4-week treatment with CsA induced typical tubulointerstitial fibrosis and increased TUNEL-positive cells. CsA induced a significant increase in BiP and caspase-12 expression peaked at 1 week, and then returned to normal levels at 4 weeks.In contrast, the expression of eIF2α, GADD153 and caspase-3 in CsA-treated rat kidneys were significantly in-creased in a time-dependent manner .CONCLUSION:Excessive endoplasmic reticulum stress causes apoptotic cell death by depleting molecular chaperones and stimulating the proapoptotic pathway in chronic CsA nephrotoxicity .