CAJ | 학술논문

　　目的探讨瘦素(leptin)与阿尔茨海默病的联系。方法将人神经纤维母细胞瘤细胞系(SH-SY5Y)分为3组：对照组、冈田酸(okadaic acid，OA)损伤模型组(40 nmol/L OA诱导12 h)和Leptin处理组。用OA诱导SH-SY5Y，建立阿尔茨海默病细胞模型。Leptin处理组是在模型组基础上，给予外源性leptin(0.4μg/ml)处理6 h，用Western Blot检测细胞周期依赖性蛋白激酶5(Cdk5)和β-actin蛋白表达水平。瘦素受体(ObR)和Cdk5的共定位情况用双重免疫荧光染色检测。结果模型诱导成功后，Western Blot显示Cdk5表达明显升高(P＜0.01)；给予leptin后，Cdk5蛋白表达显著降低(P＜0.01)。双染法免疫荧光实验表明，SH-SY5Y细胞中ObR和Cdk5蛋白均主要在细胞质中表达，并且红色和绿色标记重叠为橙色荧光，可认为两种蛋白共定位，提示ObR和Cdk5之间可能存在相互作用。结论 Leptin能够在体外降低阿尔茨海默病相关的Cdk5表达，为其治疗提供了新靶点。
　　목적탐토수소(leptin)여아이자해묵병적련계。방법장인신경섬유모세포류세포계(SH-SY5Y)분위3조：대조조、강전산(okadaic acid，OA)손상모형조(40 nmol/L OA유도12 h)화Leptin처리조。용OA유도SH-SY5Y，건립아이자해묵병세포모형。Leptin처리조시재모형조기출상，급여외원성leptin(0.4μg/ml)처리6 h，용Western Blot검측세포주기의뢰성단백격매5(Cdk5)화β-actin단백표체수평。수소수체(ObR)화Cdk5적공정위정황용쌍중면역형광염색검측。결과모형유도성공후，Western Blot현시Cdk5표체명현승고(P＜0.01)；급여leptin후，Cdk5단백표체현저강저(P＜0.01)。쌍염법면역형광실험표명，SH-SY5Y세포중ObR화Cdk5단백균주요재세포질중표체，병차홍색화록색표기중첩위등색형광，가인위량충단백공정위，제시ObR화Cdk5지간가능존재상호작용。결론 Leptin능구재체외강저아이자해묵병상관적Cdk5표체，위기치료제공료신파점。
Objective To study the association between leptin and Alzheimer's disease (AD). Methods SH-SY5Y cells were randomly divided into control group, Okadaic acid (OA) injury model group, and leptin treatment group. An AD model was established by inducing SH-SY5Y with 40nmol/L OA. Leptin treatment group was treated with 0.4μg/ml exogenous leptin for 6 h. Expression levels of cell cycle-dependent Cdk5 and β-actin proteins were measured by Western blot. ObR and Cdk5 co-localization was detected by dual immunofluorescent staining assay. Results Western blot showed that the Cdk5 expression level was significantly higher after the model was established (P<0.01) and significantly lower after leptin was given (P<0.01). Double immunofluorescence staining assay showed that the ObR and Cdk5 proteins were expressed mainly in cytoplasm of SH-SY5Y cells. The red and green markers were overlapped into orange fluorescence, which could be considered as the co-localization of the two proteins, suggesting that ObR interacts with Cdk5. Conclusion Leptin can down-regulate Alzheimer's disease-related Cdk5 expression in vitro, thus providing a new target for the treatment of Alzheimer's disease.