17β-雌二醇介导的信号通路对急性呼吸窘迫综合征小鼠肺泡上皮钠离子通道的调控研究

发布时间：2018-05-08 15:53

本文选题：呼吸窘迫综合征 + 雌二醇　；参考：《中国全科医学》2014年27期

【摘要】：目的探讨17β-雌二醇介导的PI3K/AKT/SGK1信号通路对小鼠急性呼吸窘迫综合征(ARDS)的保护作用及其对肺泡上皮钠离子通道(ENaC)的调控。方法 40只雄性C57B/L6小鼠随机分为对照组(control组)、模型组(LPS组)、17β-雌二醇治疗组(E2组)、渥曼青霉素干预组(wortmannin组),每组10只。苏木素-伊红(HE)染色观察肺组织病理改变并进行肺损伤评分;血气分析动脉血氧分压(PaO2)变化;肺湿干比(W/D)检测肺水肿的严重程度;BCA法检测肺泡灌洗液(BALF)中总蛋白含量;酶联免疫吸附试验(ELIAS)法检测BALF中白介素(IL)-1β、肿瘤坏死因子α(TNF-α)水平及髓过氧化物酶(MPO)活性;反转录聚合酶链式反应(RT-PCR)法检测各亚基ENaC mRNA表达;Western blotting法检测AKT、SGK1磷酸化水平及各亚基ENaC蛋白表达水平变化。结果小鼠气管内注射脂多糖(LPS)造模后,LPS组肺损伤评分为(4.2±0.2)分,E2组为(2.7±0.5)分,wortmannin组为(3.9±0.3)分,差异有统计学意义(F=46.07,P=0.000);其中E2组较LPS组降低,wortmannin组较E2组升高(P0.05)。4组不同时间PaO2比较有差异(F=188.148,P=0.000)。4组小鼠肺组织W/D,BALF中总蛋白含量,IL-1β、TNF-α水平,MPO活性,α-、β-、γ-ENaC的mRNA表达水平,α-、β-、γ-ENaC蛋白表达及p-AKT、p-SGK1表达水平比较有差异(P0.05);其中与control组比较,LPS组、E2组、wortmannin组W/D、BALF中总蛋白含量、IL-1β、TNF-α水平、MPO活性升高,α-、β-、γ-ENaC的mRNA表达水平降低(P0.05);与E2组比较,LPS组、wortmannin组W/D、BALF蛋白含量、IL-1β、TNF-α水平、MPO活性升高,α-、β-、γ-ENaC蛋白表达水平及p-AKT、p-SGK1表达水平降低(P0.05)。结论 17β-雌二醇可通过快速激活PI3K/AKT/SGK1信号通路介导的转录后调控机制上调ENaC表达,从而对小鼠ARDS肺水肿发挥保护作用。
[Abstract]:Objective to investigate the protective effect of 17 尾 -estradiol mediated PI3K/AKT/SGK1 signaling pathway on acute respiratory distress syndrome (ARDS) in mice and its regulation on alveolar epithelial sodium channel (ENAC). Methods Forty male C57B/L6 mice were randomly divided into control group (n = 10), control group (n = 10) and control group (n = 10). The pathological changes of lung tissue were observed by hematoxylin-eosin (HEH) staining and lung injury score was evaluated. The changes of arterial partial pressure of oxygen (Pao _ 2) were analyzed by blood gas analysis. The severity of pulmonary edema was detected by WR / D and the total protein content in alveolar lavage fluid (BALF) was detected by BCA method. Enzyme linked immunosorbent assay (Elisa) was used to detect the level of IL-1- 尾, TNF- 伪 and the activity of myeloperoxidase (MPO) in BALF. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of ENaC mRNA of each subunit. Western blotting was used to detect the phosphorylation level of AKT-1 SGK1 and the changes of ENaC protein expression in each subunit. Results after endotracheal injection of lipopolysaccharide (LPS), the lung injury score of LPS group was 4.2 卤0.2) and that of E2 group was 2.7 卤0.5). The score of wortmannin group was 3.9 卤0.3). There were significant differences in the content of total protein IL-1 尾 -TNF- 伪, the mRNA expression of 伪 -, 尾 -, 纬 -ENaC, 伪 -, 尾 -, 纬 -ENAC protein expression and the expression of 伪 -, 尾 -, 纬 -ENaC protein in lung tissue of WR DBALF of mice in E _ 2 group compared with E _ 2 group, P0.054.The difference of total protein content in lung tissue of WR / DBALF was statistically significant (P < 0.01), and the mRNA expression level of wortmannin group was lower than that of LPS group compared with that of E _ 2 group. There were significant differences in PaO2 content in lung tissue of WR / DBALF group compared with that of E _ 2 group. Compared with control group, the total protein content of WR 尾 -TNF- 伪 increased and the mRNA expression level of 伪 -, 尾 -, 纬 -ENAC decreased P0.05a. Compared with E2 group, the level of WR / DBALF protein and the activity of IL-1 尾 TNF- 伪 increased in LPS group and E2 group, respectively, and the activity of IL-1 尾 -TNF- 伪 increased in LPS group compared with that in LPS group (P < 0 05), and the expression of 伪 -, 尾 -, 纬 -ENAC protein in WR / DBALF group was significantly increased than that in LPS group (P < 0 05), while the expression of 伪 -, 尾 -, 纬 -ENAC protein in WR / DBALF group was significantly higher than that in LPS group (P < 0 05). The expression level of 伪 -, 尾 -, 纬 -ENaC protein and p-AKTna-p-SGK1 protein decreased significantly (P 0.05). Conclusion 17 尾 -estradiol can up-regulate the expression of ENaC through the mechanism of rapid activation of PI3K/AKT/SGK1 signaling pathway, and thus play a protective role against ARDS pulmonary edema in mice.
【作者单位】：重庆医科大学附属第二医院呼吸内科;
【基金】：国家自然科学基金资助项目(81270141)
【分类号】：R563.8;R-332

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