油酸在黄曲霉毒素诱导肝细胞损伤中的保护作用

为了解油酸在黄曲霉毒素B1（aflatoxin B1，AFB1）诱发的肝损伤中所起的保护作用，以体外培养的肝细胞（L-02）作为靶细胞，研究油酸的保护机制。设置3组实验：对照组（无处理）、AFB1组（只有AFB1）、油酸组（AFB1和油酸共同处理），药物处理24h后显微镜观察细胞形态，细胞增殖-毒性检测试剂盒（Cell Counting Kit-8, CCK-8）检测细胞活力，荧光法检测活性氧（reactive oxygen species，ROS）水平，Annexin V-FITC/PI检测细胞凋亡率，蛋白质免疫印迹检测血红素加氧酶-1（heme oxygenase 1，HO-1）、Caspase-3以及Survivin蛋白的表达。采用t检验或单因素方差分析进行统计分析。结果发现：与对照组相比，AFB1暴露能够明显抑制细胞活力（P<0.05），促进细胞凋亡（P<0.001），升高ROS水平（P<0.001）；与AFB1组相比，油酸作用后细胞活力显著增加（P<0.01），细胞凋亡减少（P<0.01），ROS水平降低（P<0.001）。与AFB1组相比，油酸作用后可显著提高 HO-1（P<0.05）和抑凋亡蛋白Survivin的表达（P<0.05），降低凋亡蛋白Caspase-3（P<0.01）表达。因此认为油酸对AFB1引发的肝细胞损伤具有保护作用，其机制可能与其通过促进抗氧化酶蛋白HO-1的表达，从而降低氧化应激水平，降低细胞凋亡有关。

Protective effect of oleic acid on aflatoxin-induced hepatocyte injury #br#

To investigate the protective effect of oleic acid on liver injury induced by aflatoxin B1, the in vitro cultured hepatocytes (L-02) were used and divided into 3 groups: control group, AFB1 group and oleic acid group (AFB1 and oleic acid co-treatment). After 24 hours of drug treatment, cell morphology was observed by microscope, cell viability was detected by CCK-8，ROS was detected by fluorescent probe, cell apoptosis was analyzed by flow cytometry, expression of HO-1, Caspase-3 and Survivin proteins were detected by Western blot. Results showed that AFB1 exposure significantly inhibited cell viability (P < 0.05), promoted cell apoptosis (P < 0.001) and ROS level (P < 0.001) compared with the control group. Compared with AFB1 group, cell activity significantly increased after oleic acid treatment(P <0.01), cell apoptosis(P <0.01) and ROS level were decreased （P <0.001）. The results also suggested that oleic acid significantly increased the expression of HO-1 (P < 0.05) and survivin (P < 0.05), and decreased the expression of caspase-3 (P <0.01). In summary, the results indicated that oleic acid had protective effect on AFB1-induced hepatocyte injury via alleviation of oxidative stress level and cell apoptosis by promoting antioxidant protein HO-1 expression.