摘要
目的 探讨高生长激素（GH）水平对体外培养的人脐静脉内皮细胞株（HUVEC）功能的影响以及盐酸法舒地尔对相关功能改变的干预效应.方法 体外培养HUVEC,并分为对照组、高GH组、盐酸法舒地尔组.CCK-8法检测各组细胞活力的改变,成管试验检测各组内皮细胞成管能力的差异;荧光法检测各组培养液一氧化氮（NO）含量的变化,Western blot检测内皮细胞NO合成酶（eNOS）总蛋白和eNOS（Thr495）磷酸化蛋白的变化.结果 与对照组相比,长时间的高GH水平孵育对内皮细胞增殖活力无明显抑制作用,但能抑制内皮细胞成管能力（P＜0.05）;而盐酸法舒地尔能够明显改善高GH水平处理后内皮细胞的成管能力（P＜0.05）.与对照组相比,高GH水平下内皮细胞eNOS（Thr495）磷酸化蛋白表达明显升高,释放NO明显减少（P＜0.05）;而盐酸法舒地尔能够抑制高GH水平诱导的内皮细胞eNOS（Thr495）磷酸化,从而促进NO的释放（P＜0.05）.结论 高GH水平可导致内皮细胞成管能力减弱,并通过内皮细胞eNOS（Thr495）磷酸化减少NO的生成;盐酸法舒地尔能够部分抑制高GH水平的作用,从而改善内皮细胞相应的功能.
Objective To investigate the effect of growth hormone （GH） excess on the functions of human umbilical vein endothelial cells （HUVECs） lines incubated in vitro and the intervention effect of fasudil hydrochloride on the related functional changes.Methods HUVECs were incubated in vitro,and they were divided into a control,a growth hormone excess,and a fasudil hydrochloride group.The changes of cell viability in each group were detected by CCK-8 assay,and the differences of endothelial cell tube formation ability in each group were detected by the matrigel tube formation assay.The changes of nitric oxide （NO） content in each group were detected by the fluorescence method.The changes of total protein of endothelial NO synthase （eNOS） and eNOS （Thr495） phosphorylated protein were detected by Western blot.Results Compared with the control group,a long-time GH excess level incubation did not have significant inhibitory effect for the endothelial cell proliferation activity,but it inhibited the endothelial cell tube formation ability （P 〈0.05）;while fasudil significantly improved the endothelial cell tube formation ability after GH excess level treatment （P 〈 0.05）.Compared with the control group,the protein expression of phosphorylated eNOS （Thr495） increased significantly under the GH excess level,the NO release decreased significantly （P 〈 0.05）;while fasudil inhibited the GH excess level induced eNOS （Thr495） phosphorylation,thereby promoting the release of NO （P 〈 0.05）.Conclusions The GH excess level may result in diminished the endothelial cell tube formation ability,and reduce the generation of NO through eNOS （Thr495） phosphorylation.Fasudil hydrochloride may partially inhibit GH excess level,thereby improving the corresponding function of endothelial cells.
出处
《中华神经外科杂志》
CSCD
北大核心
2015年第8期841-845,共5页
Chinese Journal of Neurosurgery
基金
国家863计划（2014AA020611）
国家自然科学基金（81172391）
