摘要
目的观察Apelin-13对同型半胱氨酸(Hcy)诱导的血管内皮细胞损伤的保护作用并探讨其可能的作用机制。方法采用10 mmol/L Hcy处理人脐静脉内皮细胞(HUVECs)24 h,诱导内皮细胞的损伤。采用不同浓度(0.1μmol/L、1.0μmol/L和10.0μmol/L)的Apelin-13预处理1 h,再加入含Hcy的培养基中继续孵育24 h,观察细胞的损伤情况。采用MTT法测定细胞活力,比色法测定细胞培养液中乳酸脱氢酶(LDH)的活性。采用Western Blot检测HUVECs中磷酸化的腺苷酸活化蛋白激酶(AMPK)蛋白表达。结果与对照组比较,Hcy组细胞的存活率显著降低,细胞培养液中LDH活力显著性升高(均P<0.05)。与Hcy组组比较,Apelin-13(1.0和10.0μmol/L)预处理组细胞的存活率均显著升高,细胞培养液中LDH活力显著性降低(均P<0.05)。Hcy组HUVECs细胞中p-AMPK蛋白的表达与对照组比较没有显著性差异(均P>0.05)。与Hcy组比较,Apelin-13(1.0和10.0μmol/L)预处理组HUVECs细胞中p-AMPK蛋白的表达显著性下调(均P<0.05)。AMPK抑制剂Compound C部分取消了Apelin-13的内皮细胞保护作用。结论 Apelin-13抑制了同型半胱氨酸诱导的内皮细胞损伤,其机制与可能与上调AMPK的磷酸化有关。
Objective To observate the protective effect of Apelin-13 on homocysteine （ Hcy）-induced damage of vascular endothelial cells and to explore the possible mechanism. Methods Human umbilical vein endothelial cells （HUVECs） were incubated with 10 mmol/L Hcy to induce damage. After pretreatment with Apelin-13 （0. 1,1.0 and 10.0 μmol/L） for 1 h,the protective effect of Apelin-13 was investigated. MTT assay was used to detect the survival rate of cells. The level of lactate dehydrogenase （LDH） in the medium was measured by colorimetry. The expression of Amp activa- ted protein kinase （AMPK） in HUVECs was measured by Western Blot. Results Compared with the control group,the survival rate of cells was singificantly decreased, the level of LDH in the medium was singificantly increased in the Hcy group （ all P 〈 0.05 ）. Compared with the Hcy group, the survival rate of cells was singificantly increased, the level of LDH in the medium was singificantly decreased in the Apelin-13 （ 1.0 and 10. 0 μmol/L） groups （ all P 〈 0.05 ）. Compared with the control group,the expressions of p-AMPK in the Hcy group did not have singificant difference （P 〉 0.05 ）. Com- pared with the Hcy group,the expressions of p-AMPK in Apelin-13 （1.0 and 10.0 μmol/L） groups were singificantly up-regulated （ all P 〈 0.05 ）. The endothelial cell protection of Apelin-13 was partly abolished by the inhibitor of AMPK Com- pound C. Conclusion Apelin-13 inhibits the damage induced by homocysteine in HUVECs,which the mechanisms may be related to the up-regulation of AMPK phosphorylation.
出处
《中南医学科学杂志》
CAS
2016年第1期29-33,共5页
Medical Science Journal of Central South China
基金
衡阳市科技计划项目(2011KJ49)
