摘要
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Sprague-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(15mg·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。
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· mg·kg^-1· d^-1） or CsA （15 mg/kg） daily for 1 or 4 weeks.Tubulointerstitial fibrosis and apoptotic cell death were estimated by trichrome staining and TUNEL staining .In addition , immunohistochemistry and immunoblotting were used to evaluate the expression of immunoglobulin -binding protein （ BiP） , eukaryotic 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 increased 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 .
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第6期1047-1051,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81160092)
关键词
环孢素A
肾毒性
内质网应激
细胞凋亡
Cyclosporine A
Nephrotoxicity
Endoplasmic reticulum stress
Apoptosis
