充血性心力衰竭患者血清对内皮祖细胞凋亡的影响及卡维地洛对凋亡的干预作用

Effects of the serum of patients with congestive heart failure on the apoptosis of endothelial progentior cells and protective role of carvedilol

目的观察充血性心力衰竭（CHF）患者血清对内皮祖细胞（EPCs）凋亡的影响及卡维地洛对凋亡的干预作用。方法选取2007年6月至2007年8月南昌大学第一附属医院心内科CHF患者20例，各抽外周血10mL获取血清用于实验。20例健康人（南昌大学第一附属医院门诊体检者）抽取外周血密度梯度离心法获取单个核细胞，贴壁法培养获得EPCs。流式细胞术及免疫荧光染色鉴定EPCs。EPCs分4组：A组加入健康人血清；B组加入CHF血清；C组加入CHF血清与卡维地洛；D组加入CHF血清与美托洛尔。继续培养24h后，膜联蛋白V／碘化丙啶（AnexinV／PI）双染色法检测EPCs凋亡率，并测EPCs的半胱胺酸蛋白酶蛋白-3（caspase3）活性。ELISA法测CHF血清中TNF-α含量，组问均数比较用方差分析及q检验。结果B组EPCs凋亡率[（7．5±0．9）％，caspase3活性（3．0±0．25）]明显高于A组[凋亡率（3．4±0．7）％，caspase3活性（1．37±0．2），P〈0．01]，c组EPCs凋亡率[（4．0±0．9）％，caspase3活性（1．54±0．12）较B组明显下降，P〈0．01]，而D组[凋亡率（7．2±1．2）％，caspase3活性2．9±0．3）较B组变化不明显，P〉0．05]。B组中EPCs凋亡率与相应CHF血清TNF-α含量呈正相关（r=O．476，P=0．03）。结论CHF患者血清能促进EPCs凋亡，卡维地洛能有效的抑制这种现象，其作用与抑制Caspase3活性有关。

Objective To investigate the effect of the serum of patients with congestive heart failure （CHF） on the apoptosis of endothelial progentior cells（EPCs） and protective role of carvedilo. Method Totally 20 patients with CHF were enrolled from Department of Cardiology of the First Hospital Affiliated of Nanchang University between June and August 2007. Serum was isolated from blood of the patients. Mononuclear cells were isolated from blood of 20 healthy volunteers by density gradient centfifugation. Attaching cells were harvested after culture for 7 clays and the EPCs were analyzed by flow cytometry and immunofluorescence method. EPCs were divided into 4 groups：group A, cultured in serum of vohmteers; group B, cultured in serum of CHF;group C, cultured in serum of CHF and Carvedilol; group D, cultured in serum of CHF and metoprolol. After culture for 24 hours, annexin V/PI was used to investigate the apoptosis rate of EPCs and the activity of caspase 3 were examined. ELISA was used to detect the level of serum TNF-a of patients with CHF. Multiple group comparison was conducted with variance analysis and q test. Results The apoptosis rate and caspase 3 activity of EPCs in group B [ （7.54- 0.9） % vs. （3.0±0.25）]were significantly increased compared with group A[（3.4 ± 0.7）% vs. （1.37 ± 0.2）, P 〈 0.01 ], and group C [ （4.0 ± 0.9） % vs. （ 1.54 ± 0.12） ]were significantly decreased compared with group B（ P 〈 0.01）. The apoptosis rate of EPCs correlated with the serum TNF-α levels of CHF patients. （r = 0.476, P = 0.03）. Conclusions The serum of patients with CHF can stimulate the apoptosis of EPCs, and carvedilol can pro- tect EPCs from apoptosis. The protective role of carvedilol may have relation with the caspase 3 activation.