Influence of RhoA-Rock pathway inhibitor on the filamentous actin of hypoxia human pulmonary microvascular endothelial cells / 中国小儿急救医学

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 groupscontrol 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.