瘦素对缺氧复氧培养L02肝细胞功能和凋亡的影响

Effect of leptin on hypoxic-reoxygenation-induced apoptosis and liver function of human L02 cells

目的观察瘦素对缺氧复氧人肝脏细胞（L02）的肝功能保护和拮抗细胞凋亡的作用。方法将L02细胞分别分为正常对照组、单纯缺氧12h复氧组和缺氧12h复氧加不同浓度的瘦素（分别为100、200、400、800、1600μg／L）干预组，用CCK-8检测各组细胞的A490值，与正常组比较算出细胞的存活率；取细胞上清液检测ALT；流式细胞仪检测细胞的凋亡率以及电镜检测细胞凋亡的形态学改变。结果与正常对照组比较，L02细胞经缺氧12h复氧培养后，CCK-8检测细胞的存活率下降（P〈0．01），加用不同浓度瘦素干预组细胞存活率较单纯缺氧12h复氧组增高（P〈0．01），以400μg/L LEP处理组细胞存活率增高明显；流式细胞仪检测细胞凋亡率结果与CCK-8法一致；电镜检测单纯缺氧复氧后细胞呈现明显凋亡形态学改变，而加用瘦素100μg／L处理细胞组细胞仅轻度改变；与正常组比较，细胞缺氧复氧后ALT升高明显（P〈0．01），加用不同浓度瘦素组处理后细胞ALT较单纯缺氧复氧组升高幅度下降（P〈0．05）。结论瘦素对缺氧复氧培养后的L02细胞肝功能有保护作用，并能减轻肝细胞凋亡。

Objective To observe the effects of leptin on hypoxic-reoxygenation-induced apoptosis and liver function of human L02 cells. Methods L02 cell damage was induced by hypoxia （95% N2 and 5% CO2 ）. The culture L02 cells were divided into hypoxic group （hypoxia alone for 12 h）, normal control group, and groups of hypoxia plus treatment with leptin （100,200,400,800 or 1600 μg/L） in vitro. In each group ,A490 values in cells were assayed with CCK-8 （ Cell Counting Kit-8 ）. The content of ALT was measured, and the apoptosis rate was assayed by flow cytometry. Results Compared with the normal control group, the survival rate of L02 cells in hypoxia group was decreased significantly （P 〈 0.01 ）. Flow eytometry revealed that apoptosis rate was significantly reduced in the leptin-treated groups as compared with the hypoxia group （ P 〈 0.05 ）. The content of ALT was increased in the hypoxia group as compared with the normal control group （P 〈 0.01 ）. In the leptin-treated groups, the content of ALT was declined as compared with the hypoxia group （ P 〈 0.05 ）. Conclusion Leptin can protect cultured L02 cells against anoxia-reoxygenation injury and the liver function induced by the anoxia-reoxygenation, and also alleviate apoptosis.