瘦素对缺氧复氧L02肝细胞凋亡及Fas／FasL表达的影响

Effects of leptin on hypoxia-reoxygenation induced apoptosis in human L02 cells and expression of Fas and FasL

目的:观察瘦素(leptin)对缺氧复氧人正常肝细胞(L02)凋亡的影响。方法:将L02细胞分别分为正常对照组、单纯缺氧12 h复氧组(IR组)和缺氧12 h复氧加不同浓度的瘦素(分别为100μg/L、200μg/L、400μg/L、800μg/L和1 600μg/L)干预组,以流式细胞仪分析、DNA缺口末端标记(TUNEL)试验、荧光定量PCR等方法观察leptin对L02肝细胞凋亡、Fas/FasLmRNA表达的影响。结果:(1)与正常对照组相比,IR组细胞凋亡率和TUNEL细胞阳性率增加(P<0.01),加用不同浓度瘦素干预组的细胞凋亡率和TUNEL细胞阳性率与IR组相比明显下降(P<0.05);(2)与正常对照组相比,IR组L02细胞中Fas/FasLmRNA表达明显上调(P<0.01);加用不同浓度瘦素干预组与IR组相比,Fas/FasLmRNA表达下降,以400μg/L瘦素作用明显,结果有显著差异(P<0.05)。结论:瘦素能减轻缺氧复氧培养L02肝细胞的凋亡,其机制可能与其下调细胞中Fas/FasLmRNA的表达有关。

AIM： To observe the effect of leptin （LEP） on hypoxia -reoxygenation induced apoptosis in L02 cells. METHODS： In the experiment, L02 cell injury was induced by hypoxic air （95% N2 and 5 % CO2 ）. The cultured L02 cells were divided into hypoxic 12 h group （IR group） alone, normal control group and the hypoxic plus leptin （100 μg/L, 200μg/L, 400 μg/L, 800μg/L and 1 600 μg/L） treatment groups in vitro. Flow cytometry, terminal dUTP nick -end labeling （TUNEL） assay and fluorescent quantitative PCR were used to measure the changes of apoptosis in L02 cells and expression of Fas/FasL mRNA. RESULTS ： （ 1 ） The percentage of L02 cells apoptosis and positive TUNEL cells significantly increased in IR group compared to control group （P 〈 0.01 ）. When L02 cells were treated with LEP, the percentage of cell apoptosis and positive TUNEL cells were decreased compared to IR group. （2） Compared to control group, the Fas/FasL mRNA expression significantly increased in IR group （P 〈0.01 ）. When L02 cells were treated with LEP, the Fas/FasL mRNA expression decreased compared to IR group, the effect of LEP at concentration of 400 μg/L was obviously （ P 〈 0.05 ）. CONCLUSION ： LEP decreases the apoptosis of hypoxic - reoxygenation L02 cells by down - regulation of Fas/FasL mRNA expression in L02 cells.