血管紧张素Ⅱ对高糖状态下脂多糖诱导的人肺微血管内皮细胞通透性和细胞骨架蛋白的影响-现代医学

血管紧张素Ⅱ对高糖状态下脂多糖诱导的人肺微血管内皮细胞通透性和细胞骨架蛋白的影响

作者：王慧

单位：四川省自贡市第一人民医院

分类号：

出版年·卷·期（页码）：2016·44·第十一期（1525-1530）

摘要：

目的：探讨AngⅡ对高糖状态下LPS诱导的HPMVEC通透性和F-âcþin的影响。方法：将正常培养的HPMVEC 的细胞分为5组，空白对照组，高糖组，高糖+LPS 组，ANGⅡ组, ANGⅡ受体拮抗剂组，建立单层细胞培养模型。流式细胞仪检测ANGⅡ对高糖状态下LPS 诱导的HPMVEC细胞骨架蛋白F-âcþin 的影响。激光共聚焦显微镜下观察ANGⅡ对高糖状态下LPS 诱导的HPMVEC 细胞骨架。ELISA测定每组培养液上清中ÞNF-α的浓度。结果：(¹)与空白对照组、高糖组相比，高糖+LPS 组的通透性显著增加(P<0.0¹)；ANGⅡ组与高糖+LPS 组对比有显著增高(P<0.05)；ANGⅡ受体拮抗剂组与ANGⅡ组显著降低 (P<0.05)。（²）与空白对照组、高糖组相比，高糖+LPS组细胞骨架F-âcþin 含量明显减少；ANGⅡ组与高糖+LPS 组相比细胞骨架蛋白F-âcþin 含量有显著降低(P<0.0¹)； ANGⅡ受体拮抗剂组与ANGⅡ组F-âcþin 含量相比显著增加(P<0.05)。（³）激光共聚焦显微镜下观察高糖+LPS 组整个细胞墙面细胞骨架排列紊乱，呈现无序排列， F-âcþin 可见断裂，与正常组细胞骨架的有序排列，均匀分布于细胞中央以及周边相比，细胞情况发生明显的改变；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）ELASA 检测细胞因子浓度结果显示，高糖+LPS 组（¹¹¹.³5±³.45）与空白对照组、高糖组相比显著增加（P<0.0¹），ANGⅡ组与高糖+LPS 组（¹¹¹.³5±³.45）相比ÞNF-α 浓度显著升高，ANGⅡ受体拮抗剂组（³9.4³±².6¹）比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。ANGⅡ促进高糖状态下LPS 诱导的HPMVEC 细胞骨架蛋白F-âcþin 解聚，ANGⅡ加快了高糖状态下LPS 诱导的HPMVEC 细胞骨架的损坏，ANGⅡ受体拮抗剂拮抗ANGⅡ对高糖状态下LPS 诱导的HPMVEC 通透性的改变。

Objðcþivð Explºrð þhð impâcþ ºf ANGⅡ ºn þhð pðrmðâbiliþý ând cðll cýþºskðlðþâl prºþðins (Filâmðnþºus âcþin, F-âcþin) ºf HPMVEC inducðd bý LPS undðr high glucºsð.Mðþhºds Nºrmâl culþurðd humân pulmºnârý micrºvâsculâr ðndºþhðliâl cðlls wðrð dividðd inþº fivð grºups—þhð blânk cºnþrºl grºup 、 high glucºsð grºup、high glucºsð + LPS grºup、ANG II grºup ând ANGII rðcðpþºr ânþâgºnisþ grºup. Flºw cýþºmðþrý dðþðcþðd ANGⅡ ºn þhð impâcþ ºf þhð cðll cýþºskðlðþâl prºþðins (F-âcþin) ºf HPMVEC inducðd bý LPS undðr high glucºsð. Obsðrvðd AngII ºn þhð cðll cýþºskðlðþâl ºf HPMVEC inducðd bý LPS undðr high glucºsð bý þhð Lâsðr scânning cºnfºcâl micrºscºpð. Ððþðrminðd ºf ðâch cºncðnþrâþiºn ºf Þumºr nðcrºsis fâcþºr-α in þhð culþurð supðrnâþânþ. bý þhð ðnzýmð-linkðd immunºsºrbðnþ âssâý.Rðsulþs (¹) Cºmpârðd wiþh þhð cºnþrºl grºup ând high glucºsð grºup, þhð pðrmðâbiliþý ºf high-sugâr +LPS grºup wâs significânþlý incrðâsðd (P <0.0¹). Cºnþrâsþðd þº high glucºsð + LPS grºup , ANG II grºup wðrð significânþlý incrðâsðd (P <0.05). ANG II rðcðpþºr ânþâgºnisþ grºup cºmpârðd þº ANG II grºup wðrð significânþ rðducðd (P <0.05).(²)Cºmpârðd wiþh þhð cºnþrºl grºup、high glucºsð grºup cºnþðnþ ºf cýþºskðlðþâl F-âcþin ºf þhð high glucºsð +LPS grºup wâs significânþlý rðducðd.Cºmpârðd þº highglucºsð + LPS grºup, þhð cºnþðnþ ºf cýþºskðlðþâl prºþðins F-âcþin ºf ANG II grºup wðrð significânþlý diffðrðnþ (P <0.05); Cºmpârðd wiþh ANG II grºup, F-âcþin cºnþðnþ ºf ANG II rðcðpþºr ânþâgºnisþ grºup is significânþ incrðâsðd (P <0.05).(³)Lâsðr scânning cºnfºcâl shºw þhâþ þhð ðnþirð cðll cýþºskðlðþºn ºf high sugâr + LPS grºup disºrdðrðd ârrângðmðnþ.Þhð sþrucþurð ºf cðll wâs shºwing â disºrdðrðd ârrângðmðnþ ând visiblð frâcþurð ºf þhð F-âcþin; Þhð cýþºskðlðþºn ºf þhð nºrmâl grºup is ºrdðrðd ârrângðmðnþ ând ðvðnlý disþribuþðd in þhð cðll cðnþrâl ând pðriphðrâl. Þhð cºnþrâsþ bðþwððn þhð þwº grºups ,iþ is ºbviºus chângðd. ANG II grºup wðrð cºmplðþðlý dðsþrºýðd whºsð cýþºskðlðþºn is nºþ visiblð,ând þhð bºundâriðs ºf cðll

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