宣肺方对内毒素诱导大鼠急性肺损伤mTOR/S6K1信号通路的影响
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国家自然科学基金项目(面上项目)(816738555);国家中医药管理局第四批全国中医(临床,基础)优秀人才研修项目(国中医药人教发【2017】24)


Effect of Xuanfei Recipe on mTOR/s6k Signal Pathway in Acute Lung Injury Induced by Endotoxin in Rats
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The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)

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    目的 观察宣肺方对内毒素诱导大鼠急性肺损伤mTOR/s6k信号通路的影响,并探讨其作用机制。方法 选取Wistar雄性大鼠30只,动物随机分为正常组、模型组、宣肺方组(高、低剂量组)和地塞米松组,尾静脉注射LPS制备ALI模型。采用ELISA法测定支气管肺泡灌洗液(BALF)中炎症因子 TNF-α、IL-1β和IL-6 含量,免疫组化测mTOR蛋白表达,Western blot 测肺组织P-mTOR蛋白表达,RT-PCR测肺泡灌洗液巨噬细胞Rps6kb1基因表达,并观察大鼠肺组织病理变化。结果 与正常组相比,模型组TNF-α、IL-1ß 、IL-6、Rps6kb1均显著升高(P <0.05),mTOR蛋白阳性面积率、p-mTOR蛋白表达显著降低(P <0.01, P <0.05);与模型组相比,地塞米松组TNF-α、IL-1ß 、IL-6含量均明显降低(P <0.05);宣肺方高剂量组TNF-α、IL-6、Rps6kb1基因表达明显降低(P <0.01, P <0.05),mTOR蛋白阳性面积率、p-mTOR蛋白表达显著升高(P <0.01, P <0.05),宣肺方低剂量组TNF-α含量明显降低(P <0.01)。HE染色显示模型组肺组织内可见局部肺出血、坏死,肺小静脉扩张、血管内白细胞数显著增多,肺间质水肿、炎细胞浸润。与模型组相比,各治疗组呈间质性肺炎的病理表现。结论 宣肺方对内毒素诱导大鼠急性肺损伤有保护作用,其机制可能与其能够下调TNF-α、IL-6、Rps6kb1基因表达和上调mTOR、P-mTOR蛋白表达有关。

    Abstract:

    Objective To observe the effect of Xuan Fei prescription on mTOR/s6k signaling pathway in rats with acute lung injury induced by endotoxin and to explore its mechanism. Methods 30 male Wistar rats were selected. The animals were randomly divided into normal control group, model control group, Xuan Fei Fang Group (high and low dose group) and dexamethasone group. The tail vein was injected with LPS to prepare ALI model. The levels of inflammatory factors TNF- a, IL-1 beta and IL-6 in bronchoalveolar lavage fluid (BALF) were measured by ELISA method. The expression of mTOR protein was detected by immunohistochemistry, P-mTOR protein expression in lung tissue was measured by Western blot, Rps6kb1 gene expression in alveolar lavage fluid was measured by RT-PCR, and the pathological changes of lung tissue were observed in rats. Results Compared with the normal group, the TNF- alpha, IL-1, IL-6 and Rps6kb1 in the model group increased significantly (P <0.05), the positive area rate of mTOR protein and the expression of p-mTOR protein decreased significantly (P <0.01, P <0.05). The expression of alpha, IL-6 and Rps6kb1 genes decreased significantly (P <0.01, P <0.05), the positive area rate of mTOR protein and the expression of p-mTOR protein increased significantly (P <0.01, P <0.05). HE staining showed that local pulmonary hemorrhage and necrosis were found in the lung tissue of the model group. The small veins of the lungs expanded, the number of white blood cells in the blood vessels increased significantly, and the pulmonary interstitial edema and inflammatory cell infiltration were observed. Compared with the model group, the treatment group showed interstitial pneumonia. Conclusions Xuan Fei Fang has protective effect on acute lung injury induced by endotoxin in rats. Its mechanism may be related to its downregulation of TNF- alpha, IL-6 , Rps6kb1 gene expression, and up regulation of mTOR and P-mTOR protein expression.

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余弘吉,杨爱东.宣肺方对内毒素诱导大鼠急性肺损伤mTOR/S6K1信号通路的影响[J].中国实验动物学报,,().

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  • 收稿日期:2018-04-17
  • 最后修改日期:2018-05-08
  • 录用日期:2018-06-11
  • 在线发布日期: 2018-11-12
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