CAJ | 학술논문

目的探讨N-乙酰半胱氨酸(NAC)对慢性环孢素A(CsA)肾毒性保护作用的分子机制.方法 ICR小鼠随机分为4组:(1)对照组:皮下注射橄榄油(1 ml·kg-1·d-1)4周；(2) NAC对照组:皮下注射橄榄油(1 ml· kg-1·d-1)和NAC(150 mg· kg-1·d-1,1 g/L溶于饮用水中)；(3)模型组:低盐饲料喂食下皮下注射CsA (30 mg· kg-1·d-1)4周建立慢性CsA肾毒性小鼠模型；(4)治疗组:低盐饲料喂食下同时给予CsA(30 mg· kg-1·d-1皮下注射)和NAC(150mg· kg-1·d-1饮用水中)4周.检测各组小鼠的体质量、肾功能；Masson染色观察肾小管间质纤维化；酶联免疫吸附法测定尿8-羟基脱氧鸟苷(8-OHdG)；免疫组化和Western印迹法分别检测抗衰老基因Klotho及其下游信号通路AKT-FoxO1的蛋白表达.结果与对照组相比,模型组小鼠肾功能下降[Scr(27.8±1.2) μmol/L比(19.0±2.2) μmol/L,P＜0.01]；肾小管间质带状纤维化；尿8-OHdG的排泄增加[(41.2±16.8) ng/d比(28.7±7.4) ng/d,P＜0.05]；Klotho蛋白表达减少[(17.8±4.5)％比(100.0±4.0)％,P＜ 0.01].与模型组比较,治疗组上述指标明显好转(均P＜ 0.05).直线相关分析示,Klotho蛋白的表达与尿8-OHdG水平(r=-0.934,P＜ 0.01)和A KT-FoxO1蛋白免疫活性(AKT:r=-0.939；FoxO1:r=-0.919,均P＜0.01)均呈负相关.结论 NAC对慢性环孢素A肾毒性具有保护作用,这种作用可能与上调Klotho基因表达、抑制AKT-FoxO1信号通路有关.
목적 탐토N-을선반광안산(NAC)대만성배포소A(CsA)신독성보호작용적분자궤제.방법 ICR소서수궤분위4조:(1)대조조:피하주사감람유(1 ml·kg-1·d-1)4주；(2) NAC대조조:피하주사감람유(1 ml· kg-1·d-1)화NAC(150 mg· kg-1·d-1,1 g/L용우음용수중)；(3)모형조:저염사료위식하피하주사CsA (30 mg· kg-1·d-1)4주건립만성CsA신독성소서모형；(4)치료조:저염사료위식하동시급여CsA(30 mg· kg-1·d-1피하주사)화NAC(150mg· kg-1·d-1음용수중)4주.검측각조소서적체질량、신공능；Masson염색관찰신소관간질섬유화；매련면역흡부법측정뇨8-간기탈양조감(8-OHdG)；면역조화화Western인적법분별검측항쇠로기인Klotho급기하유신호통로AKT-FoxO1적단백표체.결과 여대조조상비,모형조소서신공능하강[Scr(27.8±1.2) μmol/L비(19.0±2.2) μmol/L,P＜0.01]；신소관간질대상섬유화；뇨8-OHdG적배설증가[(41.2±16.8) ng/d비(28.7±7.4) ng/d,P＜0.05]；Klotho단백표체감소[(17.8±4.5)％비(100.0±4.0)％,P＜ 0.01].여모형조비교,치료조상술지표명현호전(균P＜ 0.05).직선상관분석시,Klotho단백적표체여뇨8-OHdG수평(r=-0.934,P＜ 0.01)화A KT-FoxO1단백면역활성(AKT:r=-0.939；FoxO1:r=-0.919,균P＜0.01)균정부상관.결론 NAC대만성배포소A신독성구유보호작용,저충작용가능여상조Klotho기인표체、억제AKT-FoxO1신호통로유관.
Objective To explore the renoprotective effect of N-acetylcysteine (NAC) on chronic cyclosporine A (CsA) nephrotoxicity in mice model.Methods ICR mice were randomly divided into 4 groups:normal control group [olive oil,1 ml/kg subcutaneous injection (s.c)],NAC control group (olive oil,1 ml/kg s.c plus NAC 150 mg · kg-1 · d-1 in drinking water),model group [ICR mice maintained on a salt-depleted diet (0.05％ sodium) plus CsA (30 mg · kg-1 · d-1) for four weeks]; Treatment group [ICR mice maintained on a salt-depleted diet (0.05％ sodium) plus CsA (30 mg·kg-1 · d-1 s.c) and NAC (150 mg · kg-1 · d-1 in drinking water).Basic parameters,histopathology,8-hydroxydeoxyguanosine (8-OHdG),the expression of Klotho and AKT-FoxO1 were studied.Results Compared with the normal control group,mice in model group showed deterioration in renal function [Scr (27.8± 1.2) μmol/L vs (19.0±2.2) μ mol/L,P ＜ 0.01],development of tubulointerstitial fibrosis,increased urinary 8-OHdG output [(41.2± 16.8) ng/d vs (28.7±7.4) ng/d,P＜0.05],and decreased Klotho expression [(17.8±4.5)％ vs (100.0 ±4.0)％,P ＜ 0.01].Concomitant administration of NAC significantly improved all above parameters (all P ＜ 0.05).Correlation analysis revealed that Klotho expression was negatively correlated with urinary 8-OHdG excretion (r =-0.934,P ＜ 0.01) and AKT-FoxO1 expression (AKT:r=-0.939,P＜ 0.01; FoxO1:r=-0.919,P＜ 0.01).Conclusion NAC can attenuate tubulointerstitial fibrosis in chronic CsA nephrotoxicity mice,which may be associated with the up-regulation of Klotho expression and the inhibition of AKT-FoxO1 signaling pathway.