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目的:探讨血管紧张素Ⅱ(AngⅡ)对高糖状态下脂多糖(LPS)诱导的人肺微血管内皮细胞(HPMVEC)通透性和其骨架蛋白纤维型肌动蛋白(F-actin)的影响。方法:将正常培养的HPMVEC分为空白对照组、高糖组、高糖加LPS组、AngⅡ组、AngⅡ受体拮抗剂组5组。建立单层细胞培养模型,流式细胞仪检测F-actin含量。激光共聚焦显微镜下观察HPMVEC骨架。ELISA测定培养液上清中肿瘤坏死因子α(TNF-α)浓度。结果:(1)细胞通透性高糖加LPS组较空白对照组、高糖组显著增加(P<0.01),AngⅡ组较高糖加LPS组显著增高(P<0.05),AngⅡ受体拮抗剂组较AngⅡ组显著降低(P<0.05)。(2)F-actin含量高糖加LPS组较空白对照组、高糖组明显减少,AngⅡ组较高糖加LPS组明显减少(P<0.01),AngⅡ受体拮抗剂组较AngⅡ组显著增加(P<0.05)。(3)镜下高糖加LPS组整个细胞墙面细胞骨架排列紊乱无序,Factin可见断裂,较空白对照组的排列有序、分布均匀有明显的改变;AngⅡ组细胞完全破坏,细胞骨架不可见,细胞分界不清;AngⅡ受体拮抗剂组细胞骨架尚清晰。(4)TNF-α浓度高糖加LPS组较空白对照组、高糖组显著增加(P<0.01),AngⅡ组较高糖加LPS组显著增加,AngⅡ受体拮抗剂组较AngⅡ组显著降低(P<0.01)。结论:AngⅡ引起高糖状态下LPS诱导的HPMVEC通透性增加,促进细胞骨架蛋白F-actin解聚,加快细胞骨架损坏,AngⅡ受体拮抗剂可拮抗HPMVEC通透性的改变。
Objective: To investigate the effect of Ang Ⅱ on lipopolysaccharide (LPS) -induced pulmonary microvascular endothelial cell (HPMVEC) permeability and its actin-like actin (F-actin) in high glucose conditions. Methods: Normal cultured HPMVEC were divided into 5 groups: blank control group, high glucose group, high glucose plus LPS group, Ang Ⅱ group and AngⅡ receptor antagonist group. Monolayer cell culture model was established, F-actin content was detected by flow cytometry. Laser confocal microscopy of HPMVEC scaffolds. The concentration of tumor necrosis factor alpha (TNF-alpha) in culture supernatant was measured by ELISA. Results: (1) Compared with control group and high glucose group, the cell permeability of high glucose group and LPS group were significantly increased (P <0.01), while those of Ang Ⅱ group were significantly higher than those of LPS group (P <0.05) Compared with Ang Ⅱ group, the dosage of the agent decreased significantly (P <0.05). (2) Compared with the control group and the high glucose group, the content of F-actin in LPS group was significantly lower than that in the control group and the high glucose group (P <0.01); the AngⅡ group was significantly higher than the LPS group (P <0.05). (3) Under the microscope, LPS group were disordered, the Factin was disorganized, compared with the blank control group, the arrangement was orderly and the distribution was even obviously changed; the cells in AngⅡgroup were completely destroyed, the cytoskeleton was not Can be seen, cell demarcation is unclear; Ang Ⅱ receptor antagonist group cytoskeleton is still clear. (4) Compared with the control group and the high glucose group, the TNF-αconcentration increased significantly (P <0.01), the AngⅡgroup increased significantly compared with the LPS group and the AngⅡreceptor antagonist group decreased significantly (P <0.01). CONCLUSION: Ang II induces the increase of HPMVEC permeability induced by LPS in high glucose, and promotes the depolymerization of F-actin, accelerates the cytoskeleton destruction. AngⅡ receptor antagonist can antagonize the change of HPMVEC permeability.