自噬反应在饮水型砷中毒所致肺损伤过程中的作用及机制研究

内蒙地区饮用水砷污染危害最为严重。已发现砷暴露可引起肺损伤，危害人群健康。但砷暴露致肺损伤发病机制尚未阐明。本研究拟采用SD大鼠饮用含不同浓度亚砷酸钠的水构建大鼠肺损伤模型并探讨其发病机制。染毒4w后分别测定血清和肺泡灌洗液中氧化应激反应因子和炎症因子含量；透射电镜对自噬体进行形态学观察；蛋白质印迹（WB）及免疫组化法测定肺组织中自噬相关蛋白LC3、P62、NBR1的表达，WB测定凋亡相关蛋白Caspase-3，-9表达，免疫荧光法测定LC3和Bcl-2的表达，TUNEL法检测肺组织凋亡指数。采用自噬特异性抑制剂 3-甲基腺嘌呤干预自噬发生，阐明自噬在肺损伤中作用及分子机制，并运用 NF-κB 抑制剂 PDTC 干预 NF-κB 信号通路，观察该通路对大鼠肺损伤自噬的调节，探讨 NF-κB 信号通路对自噬的调节机制，探讨饮水型砷暴露所致肺损伤发病机制，为地方性砷中毒预防和干预提供科学依据。

Inner Mongolia is one of the most severe areas of water arsenic contamination. It has been found that arsenic exposure can cause lung damage, which is harmful to human health. Yet the mechanism of arsenism is still not clear. In this study, in order to discover the mechanism of lung damage due to the exposure of arsenic in drinking water, a rat lung damage model is set up with drinking water of different concentrations of sodium arsenite. Oxidative stress factors and inflammation related factors are measured respectively in serum and BALF after the rats are exposure to arsenic for 4w, and lung damage is graded as well. Mophological observation on autophagosome is undertaken using transmission electron microscope; Autophay related protein LC3, P62, NBR1 are measured by Western blotting and immunohistochemistry. Apoptosis related protein expression of Caspase-3 and -9 are measured by Western blotting. The expression of LC3 and Bcl-2 are measured by immunohistochemistry. Lung apoptotic index of rats is detected. Autophagy is intervened by applying autophagy specific inhibitor of 3-MA, which can preliminary clarify the role of autophagy in lung damage and its molecular mechanism. And PDTC, NF-κB inhibitor, is used to intervene NF-κB signal channel to observe the regulation of NF-κB signal channel on rat lung damage autophagy. Regulation mechanism of NF-κB signal channel is explored. The clarification of lung damage mechanism of exposure of arsenic in drinking water provides theoretical evidence for prevention and intervention of local arseniasis.