地塞米松对海水浸泡人肺泡上皮细胞钠通道的影响

发布时间：2019-03-05 15:13

【摘要】：目的探讨地塞米松对海水浸泡的人肺泡上皮细胞(A549)钠通道的影响。方法将常规培养的A549细胞分为海水处理组(SG)、阴性对照组(NG)和地塞米松处理组(DG)。SG组经灭菌配方海水常规孵育,DG组在经海水处理后,加入地塞米松(终浓度1μmol/L)处理,NG组常规培养至实验终点。配方海水渗透压1300mmol/L,pH8.2,相对密度1.05。采用MTT法检测细胞存活率,流式细胞仪检测细胞的凋亡和坏死情况,Western blotting法检测钠钾泵α1亚单位(NKA-α1)、钠通道α亚单位(α-ENaC)的表达,水解比色法检测NKA活性。结果流式细胞术检测结果显示,海水孵育30min后A549细胞的存活率为90.60%±2.11%,地塞米松(1μmol/L孵育6h)处理对海水孵育后细胞的存活率无影响。NG组的NKA-α1和α-ENaC蛋白表达(0.113±0.041、0.396±0.124)显著高于SG组(0.038±0.009、0.093±0.038;P0.01)或DG组(0.067±0.015、0.165±0.078;P0.01),但DG组显著高于SG组(P0.05)。与NG组[31.298±5.779μmol/(mg·h)]比较,SG组NKA酶活性[14.211±3.885μmol/(mg·h)]明显降低(P0.05);DG组的NKA酶活性[18.641±1.921μmol/(mg·)]较SG组有升高(P0.05)。结论在不影响人肺泡上皮细胞存活状况的情况下,地塞米松可以抑制海水孵育导致的α-ENaC、NKA-α1含量及NKA活性的下降。
[Abstract]:Objective to investigate the effect of dexamethasone on sodium channel in human alveolar epithelial cells (A549) after seawater immersion. Methods A549 cells were divided into two groups: seawater treatment group (SG),-negative control group, (NG) and dexamethasone treatment group (DG). SG group) were incubated with sterilized formula seawater. DG group was treated with seawater. After treatment with dexamethasone (1 渭 mol / L), the NG group was cultured to the end of the experiment. The osmotic pressure of the formulated seawater is 1300 mmol / L, pH 8.2, and the relative density is 1.05. Cell viability was detected by MTT method, apoptosis and necrosis were detected by flow cytometry., Western blotting assay was used to detect the expression of sodium potassium pump 伪 1 subunit (NKA- 伪 1) and sodium channel 伪 subunit (伪-ENaC). The activity of NKA was detected by hydrolysis colorimetry. Results the results of flow cytometry showed that the survival rate of A549 cells incubated with seawater was 90.60% 卤2.11%, and the survival rate of A549 cells was 90.60% 卤2.11%. The expression of NKA- 伪 1 and 伪-ENaC protein in NG group (0.113 卤0.041, 0.396 卤0.124) was significantly higher than that in SG group (0.038 卤0.009, 0.093 卤0.038). P0.01) or DG group (0.067 卤0.015,0.165 卤0.078), but DG group was significantly higher than SG group (P0.05). Compared with NG group [31.298 卤5.779 渭 mol / (mg 路h)], NKA activity in SG group [14.211 卤3.885 渭 mol / (mg 路h)] was significantly decreased (P0.05). The activity of NKA in DG group [18.641 卤1.921 渭 mol / (mg)] was higher than that in SG group (P0.05). Conclusion Dexamethasone can inhibit the decrease of 伪-ENaC,NKA- 伪 1 content and NKA activity induced by seawater incubation without affecting the survival of human alveolar epithelial cells.
【作者单位】：海军总医院呼吸内科;解放军总医院胸外科;
【基金】：全军“十一五”后勤科研计划课题(06-3305)
【分类号】：R96;R83

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