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关心康对大鼠缺血心肌细胞中ATP敏感性钾通道亚基Kir6.1、Kir6.2、SUR2A和SUR2B表达的影响

[Effects of Guanxinkang on expressions of ATP-sensitive potassium channel subunits Kir6.1, Kir6.2, SUR2A and SUR2B in ischemic myocytes of rats].

作者信息

Chen Fu-rong, Zhang Na, Liu Ping, Zhang Yi-yi, Han Xiang-hui, Cai Jue-feng

机构信息

Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.

出版信息

Zhong Xi Yi Jie He Xue Bao. 2010 May;8(5):458-64. doi: 10.3736/jcim20100510.

Abstract

OBJECTIVE

To observe the effects of Guanxinkang injection, a compound traditional Chinese herbal medicine, on ATP-sensitive potassium (K(ATP)) channel subunits in ischemic myocardial cells of rats, and to explore the mechanism of Guanxinkang in protecting myocardial ischemic reperfusion injuries.

METHODS

Forty-eight Wistar rats were randomly divided into normal group, untreated group, glibenclamide group, pinacidil group, Guanxinkang group and Guanxinkang plus glibenclamide group. The ventricular myocytes were prepared from hearts of normal rats by enzymatic dissociation method. The ischemic ventricular myocytes underwent perfusion with normal Tyrode solution for 10 min, then stopping perfusion 30 min, and followed by 45 min of reperfusion. The glibenclamide, pinacidil and Guanxinkang were added into ventricular myocytes solution directly. Then the solutions were placed at 4 degrees centigrade. After 24-hour freezing at -80 degrees centigrade, mRNA and protein expressions of KATP subunits Kir6.1, Kir6.2, SUR2A and SUR2B were measured by real-time quantitative polymerase chain reaction and Western blotting respectively.

RESULTS

In normal rat myocardial cells, there were SUR2A, Kir6.1, and Kir6.2 protein and gene expressions but no expression of SUR2B protein. In the untreated group, all subunit mRNA and protein expressions of KATP increased to some extent as compared with the normal group. Pinacidil, a potassium channel opener, significantly increased mRNA and protein expressions of KATP subunits, while the blocker glibenclamide had a reverse effect. Meanwhile, Guanxinkang injection significantly increased mRNA and protein expressions of K(ATP) subunits but with no significant difference as compared with pinacidil.

CONCLUSION

Guanxinkang injection can obviously enhance the open of KATP channel and thus play a role in cardiovascular protection.

摘要

目的

观察复方中药冠心康注射液对大鼠缺血心肌细胞三磷酸腺苷敏感性钾(K(ATP))通道亚单位的影响,探讨冠心康保护心肌缺血再灌注损伤的机制。

方法

将48只Wistar大鼠随机分为正常组、未处理组、格列本脲组、吡那地尔组、冠心康组和冠心康加格列本脲组。采用酶解法从正常大鼠心脏制备心室肌细胞。对缺血心室肌细胞用正常台氏液灌注10分钟,然后停止灌注30分钟,接着再灌注45分钟。将格列本脲、吡那地尔和冠心康直接加入心室肌细胞溶液中。然后将溶液置于4摄氏度。在-80摄氏度冷冻24小时后,分别用实时定量聚合酶链反应和蛋白质印迹法检测KATP亚单位Kir6.1、Kir6.2、SUR2A和SUR2B的mRNA和蛋白表达。

结果

在正常大鼠心肌细胞中,有SUR2A、Kir6.1和Kir6.2蛋白及基因表达,但无SUR2B蛋白表达。在未处理组,与正常组相比,KATP的所有亚单位mRNA和蛋白表达均有不同程度增加。钾通道开放剂吡那地尔显著增加KATP亚单位的mRNA和蛋白表达,而阻滞剂格列本脲则有相反作用。同时,冠心康注射液显著增加K(ATP)亚单位的mRNA和蛋白表达,但与吡那地尔相比无显著差异。

结论

冠心康注射液能明显增强KATP通道开放,从而发挥心血管保护作用。

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