卡维地洛减轻体外循环诱导的犬心脏细胞凋亡：Fas/FasL和半胱天冬酶-3途径的调控

Carvedilol attenuates CPB-induced apoptosis in dog heart: regulationof Fas/FasL and caspase-3 pathway.

作者信息

Zhang Shunye, Sun Zongquan, Liu Lixin

机构信息

Department of Cardiovascular Surgery, Shanxi Cardiovascular Hospital, Taiyuan 030024, China.

出版信息

Chin Med J (Engl). 2003 May;116(5):761-6.

PMID:12875697

Abstract

OBJECTIVE

To evaluate the effects of Carvedilol on cardiopulmonary bypass (CPB)-induced myocardiocyte apoptosis and its effects on regulation of Fas, FasL expression, caspase-3 activity and oxidative stress in the left ventricle (LV) in this setting.

METHODS

Ten adult dogs undergoing conventional hypothermic CPB were randomly divided into control and Carvedilol treated groups (n = 5, respectively). Dogs in Carvedilol treated group received a bolus of Carvedilol (1 mg/kg) intravenously and a maintenance dosage of Carvedilol (3 micro g.min(-1).kg(-1)) for 3 hours after the reperfusion of the heart. Dogs in control group received no Carvediolol. LV samples were obtained before, during and 3 hours after CPB. In situ nick end-labeling (TUNEL) technique was used to detect the apoptotic cells. The expressions of Fas and FasL were detected immunohistochemically and quantified by fluorescence activated cell sorting (FACS). The activity of caspase-3 enzyme and malondialdehyde (MDA) level were measured by cleavage of Z-DEVD-AMC substrate and thiobarbituric acid reactive substance (TBARS) method, respectively.

RESULTS

Before and during CPB, all the parameters were not significantly different intra- or between groups (P > 0.05). After CPB, these parameters in both groups were significantly elevated compared with those of before and during CPB (P < 0.028, respectively). However, the number of apoptotic cells in Carvedilol treated group was significantly decreased compared with that of the control group (P < 0.021). The expressions of Fas and FasL were significantly downregulated by Carvedilol (P < 0.001 and 0.003, respectively). The caspase-3 activity and the content of MDA in the Carvedilol treated group was also significantly reduced (P < 0.026 and 0.005, respectively).

CONCLUSIONS

Carvedilol significantly reduces CPB-induced cardiomyocyte apoptosis in dog hearts and the reduction of cardiomyocyte apoptosis is associated with downregulation of Fas and FasL expression, inhibition of caspase-3 activity and oxidative stress in LV.

摘要

目的

评估卡维地洛对体外循环（CPB）诱导的心肌细胞凋亡的影响，以及在此情况下其对左心室（LV）中Fas、FasL表达调控、半胱天冬酶-3活性和氧化应激的影响。

方法

将10只接受常规低温CPB的成年犬随机分为对照组和卡维地洛治疗组（每组n = 5）。卡维地洛治疗组的犬在心脏再灌注后静脉注射一剂卡维地洛（1 mg/kg），并给予卡维地洛维持剂量（3μg·min⁻¹·kg⁻¹），持续3小时。对照组的犬未接受卡维地洛治疗。在CPB前、CPB期间和CPB后3小时获取LV样本。采用原位缺口末端标记（TUNEL）技术检测凋亡细胞。通过免疫组织化学检测Fas和FasL的表达，并通过荧光激活细胞分选（FACS）进行定量分析。分别采用Z-DEVD-AMC底物裂解法和硫代巴比妥酸反应物质（TBARS）法测定半胱天冬酶-3酶活性和丙二醛（MDA）水平。

结果

在CPB前和CPB期间，组内和组间所有参数均无显著差异（P > 0.05）。CPB后，两组的这些参数均较CPB前和CPB期间显著升高（分别为P < 0.028）。然而，卡维地洛治疗组的凋亡细胞数量较对照组显著减少（P < 0.021）。卡维地洛显著下调Fas和FasL的表达（分别为P < 0.001和0.003）。卡维地洛治疗组的半胱天冬酶-3活性和MDA含量也显著降低（分别为P < 0.026和0.005）。