Api6在高脂高胆固醇饮食引起肺部炎症中所发挥作用的研究

Role of Api6 in lung inflammation induced by high fat /cholesterol food in C57BL/6J mice

的：研究凋亡抑制园子6（Api6）在高脂高胆固醇饮食所致C57BL/6J小鼠肺部炎症反应中的作 用。方法：6-8周龄的C57BLl6J雄性小鼠喂养于SPF环境中，随机分成2组，分别给予普通饮食和高脂高胆固醇 饮食喂养。喂养16周后收集肺组织并采用免疫组织化学和ELISA法鉴定肺组织的炎症状态。实时定量PCR和Western blotting鉴定Api6 mRNA与蛋白的表达水平，流式细胞术检测小鼠支气管肺泡灌洗液细胞凋亡情况。体外培养H噬细胞RAW264.7，流式细胞术检测Api6对氧化型低密度脂蛋白（ oxLDL）引起的细胞凋亡的影响。结果：高脂高胆固醇饮食喂养小鼠16周后，C57BL/6J小鼠肺组织出现以巨噬细胞蓄积以及肿瘤坏死因子α和单核细胞趋化蛋白1升高为主的炎症反应。与普通饮食组相比，高脂高胆固醇饮食喂养小鼠肺组织的Api6mRNA和蛋白表 达水平都显著上调（P〈0.01），同时支气管肺泡灌洗液中的巨噬细胞凋亡水平明显下降（P〈0.01）。体外实验证实500μg/L的重组Api6处理RAW264.7细胞可显著抑制oxLDL引起的细胞凋亡（P〈0.05）。结论：高脂高胆固醇饮食可致C57BL/6J小鼠肺组织巨噬细胞蓄积，其机制可能与Api6抑制巨噬细胞的凋亡有关。

AIM ： To investigate the function of apoptosis inhibitor 6 （ Api6 ） in lung inflammation induced by high-fat high-cholesterol diet （HFD/HCD） in male C57BL/6J mice. METHODS： Male C57BL/6J mice （6-8 weeks old） were randomly divided into 2 groups and treated with regular diet and HFD/HCD, respectively. After 16 weeks of feeding, the lung tissues were collected and the pulmonary inflammatory status was determined by immunohistochemistry and ELISA. The mRNA and protein expression levels of Api6 were determined by real-time PCR and Western blotting. The apoptotic rate of bronchioalveolar lavage cells was examined by flow cytometry. RAW264.7 cells were cultured in vitro and the apoptosis induced by oxidized low-density lipoprotein （oxLDL） was detected by flow cytometry. RESULTS： Accumulation of maerophages and increases in both tumor necrosis factor α and monocyte chemoattractant protein 1 were observed in the lung tissues of 16-week HFD/HCD-fed C57BL/6J mice. Compared with the regular diet-fed mice, the expression of Api6 at mRNA and protein levels in the lung tissues was highly increased in the HFD/HCD-fed mice （P 〈0.01 ）. Meanwhile, the apoptotic rate of bronchioalveolar lavage macrophages fi＇om the HFD/HCD-fed mice was highly inhibited （ P 〈 0. 01 ）. In vitro, 500 μg/L recombinant Api6 significantly inhibited the apoptosis of RAW264.7 ceils induced by oxLDL （P 〈0.05）. CONCLUSION： HFD/HCD feeding results in the accumulation of macrophages in the lung of C57BL/6J mice, which may partly due to the increased expression of Api6 and its anti-apoptotic role in macrophages.