摘要
目的探讨肝细胞生长因子(HGF)对缺氧损伤的人肺微血管内皮细胞(HPMECs)的保护作用,提高肺血管内皮细胞对缺氧的耐受性,增加损伤后的存活率,改善其功能,为防止缺氧导致的肺动脉高压提供实验依据。方法体外培养HPMECs,用物理方法建立缺氧损伤模型。实验分为空白对照组、缺氧损伤组、HGF组、PHA665752组(HGF抑制剂组)。采用免疫荧光法鉴定第7代HPMECs,四甲基偶氮唑蓝(MTT)比色法检测细胞存活率,倒置显微镜下计数在不同干扰条件下HPMECs的贴壁细胞数量,Western blot法检测细胞间黏附分子-1(ICAM-1)表达情况。结果缺氧损伤组细胞贴壁率下降,ICAM-1的表达量明显高于空白对照组(P<0.01);HGF组细胞存活率较缺氧损伤组明显提高(P<0.01),细胞贴壁率较缺氧损伤组明显增加(P<0.01),ICAM-1的表达量明显下降(P<0.01);PHA组细胞存活率较缺氧损伤组和HGF组均有明显差异(P<0.01),细胞贴壁率较缺氧损伤组明显下降(P<0.01),ICAM-1的表达明显上升(P<0.01)。结论 HGF可能是通过增加HPMECs的存活率、提高体外培养时细胞的贴壁率、减少ICAM-1表达,减轻缺氧损伤所造成的HPMECs损伤。
Objective To investigate the protective effects of hepatocyte growth factor （HGF） on hypoxic human pulmonary microvascular endothelial cells （HPMECs）. Methods HPMECs were cultured in vitro, and the hypoxic model was established by the physical method. Cells were divided into 4 groups： the control group, the hypoxic group, HGF group, and phytohemagglutinin （PHA） group. The 7th generation of HPMECs was evaluated by the method of immunocytochemistry. The persistence rate of HPMECs was measured by MTT assay and the adhesive cells were counted by the microscopy. The expression of intercellular adhesion molecule-1 （ICAM-1） protein was determined by immunofluorescence staining. Results The adherence percentage of cells significantly decreased after hypoxia, whereas the expression of the ICAM-1 protein was significantly higher in the hypoxia group than in control group （P 〈 0.01 ）. Compared with the hypoxia group, the persistence and adherence percentage of ceils in the HGF group significantly increased （ P 〈 0.01 ）, whereas the expression of the ICAM-1 protein significantly dropped （ P 〈 0.01 ）. In the PHA group, the persistence and adhesion rate were significantly different from those in the hypoxia group and HGF group （P 〈 0.01 ）, and the expression of the ICAM-1 protein increased significantly （ P 〈 0.01 ）. Conclusion HGF could inhibit the hypoxic damage of HPMECs by decreasing the persistence and the adhesive capacity of these cells and inducing expression of ICAM-1.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2013年第1期1-5,共5页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(81000018)
解放军总医院苗圃基金重点项目(10KMZ04)
航天医学基础与应用国家重点实验室开放基金(SMFA11K02)
武器预研重点基金(9140A26040312JB10078)~~
关键词
缺血性肺动脉高压
肝细胞生长因子
人肺微血管内皮细胞
细胞间黏附分子-1
hypoxic pulmonary hypertension
hepatocyte growth factor
human pulmonary microvascular endothelial cells
intercellular adhesion molecule-1
