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余弘吉,杨爱东,李小茜,吴中华,符胜光,许明亮.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,2018,26(4):431~436.
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
Effect of Xuanfei Formula on mTOR / S6K1 signal pathway in acute lung injury induced by lipopolysaccharide in rats
投稿时间:2018-04-17  
DOI:10.3969/j. issn. 1005 - 4847. 2018. 04. 004
中文关键词:  急性肺损伤  宣肺方  p?mTOR  RPS6KB1  大鼠
英文关键词:acute lung injury  Xuanfei formula  p?mTOR  RPS6KB1  rats
基金项目:国家自然科学基金面上项目(No. 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);上海中医药大学学科能力提升项目(No. A1?Z183020110);上海中医药大学高水平大学建设高峰高原团队 (No. A?U18205010127)
作者单位E-mail
余弘吉 上海中医药大学基础医学院经方理论应用研究中心,上海 201203  
杨爱东 上海中医药大学基础医学院经方理论应用研究中心,上海 201203 aidongy@126. com 
李小茜 上海中医药大学基础医学院经方理论应用研究中心,上海 201203 lixiaoqian5258@126. com 
吴中华 上海中医药大学基础医学院经方理论应用研究中心,上海 201203  
符胜光 上海中医药大学基础医学院经方理论应用研究中心,上海 201203  
许明亮 上海中医药大学基础医学院经方理论应用研究中心,上海 201203  
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中文摘要:
      目的 观察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,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?地塞米松组和宣肺方组(高?低剂量组),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 与正常组比较,模型组TNF - α?IL -1β?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);与模型组比较,地塞米松组TNF - α?IL -1β?IL -6 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;与模型组比较,各治疗组呈轻度间质性肺炎?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 含量?RPS6KB1 基因表达和上调mTOR?p?mTOR 蛋白表达有关?
英文摘要:
      Objective To observe the effect of a Chinese herbs, Xuanfei formula, on mTOR/ S6K1 signaling pathway in rats with acute lung injury induced by endotoxin, and to explore its mechanism. Methods Thirty male Wistar rats were selected and randomly divided into normal control group, model control group, dexamethasone group and XuanfeiActa Lab Anim Sci Sin,August 2018,Vol. 26. No. 4 formula group (high and low dose groups). Lipopolysaccharide (LPS) was injected into the tail vein to prepare rat model of acute lung injury (ALI). The levels of inflammatory factors TNF - α, IL - 1β and IL - 6 in bronchoalveolar lavage fluid (BALF) were detected by ELISA assay. The expression of mTOR protein was detected by immunohistochemistry, p?mTOR protein expression in the lung tissues was measured by western blot, RPS6KB1 gene expression in alveolar lavage fluid was measured by RT?PCR, and the histological changes of rat lung tissues were examined by pathology. Results Compared with the normal group, the TNF - α, IL -1β, IL -6 and RPS6KB1 in the model group were significantly increased ( P <0. 05), and the positive area ratio of mTOR protein and the expression of p?mTOR protein were significantly decreased ( P < 0. 01, P < 0. 05). Compared with the model group, the level of TNF - α, IL - 1β and IL - 6 were significantly decreased ( P < 0. 05) in the dexamethasone group ( P < 0. 05), and the level of TNF - α, IL - 6 and RPS6KB1gene expression were significantly decreased ( P < 0. 01, P < 0. 05), and the positive area ratio of mTOR protein and the expression of p?mTOR protein in the Xuanfei formula groups were significantly increased ( P < 0. 01, P < 0. 05). Pathological examination of the lung tissues of the model group showed local pulmonary hemorrhage and necrosis, dilation of small pulmonary veins, increased number of leucocytes in the blood vessels, pulmonary interstitial edema and inflammatory cell infiltration in the lung tissues. Compared with the model group, the treatment groups showed mild interstitial pneumonia. Conclusions Xuanfei formula has a protective effect on acute lung injury induced by endotoxin in rats. Its mechanism may be related to its downregulation of TNF - α, IL -6, RPS6KB1 gene expression, and upregulation of mTOR and p?mTOR protein expression.
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