Influence of RhoA-Rock pathway inhibitor on the filamentous actin of hypoxia human pulmonary microvascular endothelial cells
10.3760/cma.j.issn.1673-4912.2012.01.024
- VernacularTitle:RhoA-Rock信号通路抑制剂对缺氧人肺微血管内皮细胞丝状肌动蛋白细胞骨架的影响
- Author:
Feitong ZHANG
;
Qiliang CUI
;
Jing MO
- Publication Type:Journal Article
- Keywords:
Pulmonary hemorrhage;
Pulmonary microvascular endothelial cell;
Filament actin;
RhoA-Rock pathway;
Infant,newborn
- From:
Chinese Pediatric Emergency Medicine
2012;19(1):67-70
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the control factor of pulmonary microvascular endothelial cells in pulmonary hemorrhage with the RhoA-Rock pathway inhibitors.MethodsHuman pulmonary rnicrovascular endothelial cells were conventionally cultured,and were divided into four groups:control group,inhibitor group,hypoxia group and hypoxia group with inhibitor.As different fluorescein lsothiocyanate-phalloidin and filamentous actin (F-actin) in cytoplasm combined,it issued red fluorescence.We observed the dynamic changes of F-actin by laser scanning confocal microscope in hypoxia human pulmonary microvascular endothelial cells and recorded the value of fluorescence.ResultsThe mean fluorescence intensity of F-actin of hypoxia group in 1 h,12h and 24 h was (64.3 ±5.5)％,(60.3±4.2)％,and (47.8 ±4.6)％ as compared with the control group;the ratio of hypoxia group with inhibitor was (66.2 ±3.2)％,(67.1 ±6.2)％,and (72.5 ± 6.1 ) ％ as compared with the control group.The mean fluorescence intensity of F-actin decreased obviously after 1 h hypoxia treated to cells,decreasing to (64.3 ± 5.5 ) ％ of the control group (P ＜0.05 ) ;to 24 h,decreasing to (47.8 ±4.6) ％ of the control group(P ＜0.05).The mean fluorescence intensity of F-actin decreased to (66.2 ± 3.2) ％ of the control group after 1 h hypoxia treated in inhibitor group,which was more than 1 h in the hypoxic group.F-actin decreased obviously to (72.5 ± 6.1 ) ％ of the control group after 24 h hypoxia treated in inhibitor group.There was significant difference comparing with the hypoxia group after 24 h hypoxia(P ＜0.05).The mean fluorescence intensity of F-actin of inhibitor group without anoxic was invariant comparing with the control group(P ＞0.05).Cortex-like structure disappeared and the stress fibers arranged disorderly after hypoxia.Actin depolymedzated and broke gradually with the extension of hypoxia time.If to be hypoxic after pretreated with RhoA-Rock pathway inhibitor,cortex-like structure by the composition of pednuclear F-actin reappeared,distribution and arrangement of stress fibers in the cytoplasm tended to rule.ConclusionThe RhoA-Rock pathway mediates the damage on F-actin of pulmonary microvascular endothelial cells after hypoxia.Interfering with the RhoA-Rock pathway inhibitors can provide a new direction for the treatment of neonatal pulmonary hemorrhage.
