肝细胞生长因子抑制糖基化终产物诱导人脐静脉内皮细胞的凋亡作用

目的探讨肝细胞生长因子(HGF)抑制糖基化终产物(AGEs)诱导人脐静脉内皮细胞凋亡的作用及其相关分子机制。方法体外培养ECV-304人脐静脉内皮细胞,采用噻唑蓝(MTT)法测定HGF对AGEs作用后ECV-304细胞生长抑制率的影响;通过Hoechst33258荧光染色观察细胞形态学改变、流式细胞术测定AnnexinV-FITC/PI双染标记的细胞凋亡率,检测HGF对AGEs诱导ECV-304细胞凋亡的影响;Western印迹法检测Bax、Bcl-2蛋白的表达。结果HGF能明显降低AGEs对ECV-304细胞生长的抑制作用;AGE诱导培养的ECV-304细胞出现明显的凋亡形态学改变,在一定浓度范围内,ECV-304细胞凋亡率与AGEs的浓度和作用时间呈依赖关系,加入HGF处理后可显著降低不同时间的内皮细胞凋亡率;HGF作用ECV-304细胞后Bcl-2蛋白表达明显升高,而Bax蛋白表达无明显变化。结论AGEs能诱导内皮细胞凋亡,而HGF能部分抑制AGEs诱导的内皮细胞凋亡,其作用机制可能与上调Bcl-2蛋白的表达水平有关。

HEPATOCYTE GROWTH FACTOR INHIBITS ADVANCED GLYCOSYLATION END PRODUCTS-INDUCED APOPTOSIS IN HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS

Objective To investigate the inhibiting effect of hepatocyte growth factor (HGF)on endothelial cell apoptosis induced by advanced glycosylation end products(AGEs)and to explore its possible mechanism. Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro. The effect of HGF on the growth inhibiting rates of HUVECs with various concentrations of AGE in different time pariods(12, 24, 48, and 72 hours)was measured by methyl thiazolyl tetrazolizm(MTT)assay, the early stage apoptosis was detected with Annexin V-FITC/PI double labeled assay, the morphology of cell apoptosis was observed by hoechst33258 fluorescence staining, and the expression of apoptosis-associated genes Bax and Bcl-2 was detected by western blotting. Results Morphological observation indicated that high concentrations of AGEs induced marked apoptotic changes in HUVECs. Within a certain concentration range, AGEs induced HUVEC apoptosis in a dose- and time-dependent manner. HGF significantly inhibited the apoptosis of HUVECs induced by AGEs(P<0.05). High AGEs significantly increased Bax protein, but not Bcl-2, whereas HGF significantly increased Bcl-2 expression without affecting Bax level. Conclusion AGEs can induce the apoptosis of endothelial cells in vitro. HGF effectively attenuates high AGE-induced apoptosis through upregulating Bcl-2 gene expression.