Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (5): 458-464.doi: 10.3736/jcim20100510

• Original Experimental Research • Previous Articles     Next Articles

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

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

  1. Science and Technology Agency, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2009-12-23 Accepted:2010-03-29 Online:2010-05-20 Published:2010-05-15
  • Contact: Ping Liu


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

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 ℃. After 24-hour freezing at –80 ℃, 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.

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 KATP subunits but with no significant difference as compared with pinacidil.

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

Key words: Chinese patent drugs, Myocytes, Cardiac, Reperfusion injury, ATP-sensitive potassium channel

Table 1

Reactive system for real-time fluorescent quantization PCR"

Content Volume (juL)
h2o 9. 5
SYBR Premix Ex Taq (2X) 12. 5
Forward primer (5 jumol/L) 1
Reverse primer (5 jumol/L) 1
RT product 1
Total 25

Table 2

Effects of Guanxinkang on current though KATP channels in ventricular myocytes of rats (x±s, pA)"

Group n Current
Normal 8 83. 51±23. 67*
Untreated 8 211.45±33.67
Glibenclamide 8 129. 68±30. 92*
Pinacidil 8 317. 87±33.75*
Guanxinkang 8 313. 75±21.82a
Guanxinkang plus glibenclamide 8 199. 43±20. 02a

Table 3

Relative expression levels of KATP subunits Kir6.1, Kir6.2, SUR2A and SUR2B mRNAs in myocardial cells in different groups (x±s)"

Group n Kir 6. 1 mRNA Kir6. 2 mRNA SUR2A mRNA SUR2B mRNA
Normal 8 1.327 6±0. 278 6 1.275 3±0. 295 9 0. 957 2±0. 151 8 1.093 4±0. 228 6
Untreated 8 3. 147 7±0. 577 7* 2. 607 1±0. 358 7* 1. 795 1±0. 267 7* 2. 103 7±0. 406 0*
Glibenclamide 8 1. 575 3±0. 304 9AD 1. 176 6±0. 179 1A° 1.052 8±0. 151 2Aa 1. 075 4±0. 199 8Aa
Pinacidil 8 5. 636 1±0. 825 3. 793 6±0. 552 0*△ 3. 095 9±0. 545 3. 476 3±0. 709
Guanxinkang 8 5. 388 4±0. 950 3. 688 2±0. 509 6*aa 2. 944 0±0. 322 3. 093 8±0. 544
Guanxinkang plus glibenclamide 8 3. 042 4±0. 478 2. 510 7±0. 277 1. 935 7±0. 277 1. 965 6±0. 383

Table 4

Relative expression levels of KATP subunits Kir6.1, Kir6.2, SUR2A and SUR2B proteins in myocardial cells in different groups"

Group n Kir6,1 protean C2±s) Kir6,2 protean C2±s) SUiR^Aprotein (3土 s) SUR2B protein M (〇26,Q? 6)
Normal 8 0,332 9 士0,1S7 4 0,044 8 土0,033 3 0,117 5 土0,132 0 0
Untreated 8 1,108 0 士0,071 4 ? 0, SIS 3 土0,164 2 ? 0,751 7 土0,124 8* 0 (0, 0,000 9)
Glibenckmide 8 0,384 2 士0,484 SAD 0,016 3 土0,027 SAD 0,117 9 土0,086 7A〇 0
Finacidil 8 1,471 3 土0,300 4 1,116 5 土0,134 0?&▲ 0,859 1 土0,172 6 1,S41 8 (1,312 9, 2,129 8) ?么▲
Guansankan^ 8 2,252 2±0,474S*AAa 1,395 8 土0,290 2 1,163 3 土0,333 1,010 5 (0,956 8, 1,308 9) ?么▲
Guansiiikflng plus glibenckmide 8 1,644 9 土0,191 3 0,502 1 土0,217 3 ?▲a 0,535 0 土 0,128 1 da 0 (0, 0.000 7)n

Figure 1

Expressions of KATP subunits Kir6.1, Kir6.2, SUR2A and SUR2B proteins in myocardial cells in different groups 1: Normal group; 2: Untreated group; 3: Guanxinkang group; 4: Guanxinkang plus glibenclamide group; 5: Glibenclamide group; 6: Pinacidil group."

[1] Bai JH, Lu QJ . The role targets of anti-ischemic drugs[J]. Guo Ji Xin Xue Guan Bing Za Zhi, 2006,33(2):125-128
白纪红, 吕秋军 . 抗心肌缺血药物的作用靶点[J]. 国际心血管病杂志, 2006,33(2):125-128
[2] Liu P, Zhang YY, Zhang JS . Effects of compound prescription on soluble intercellular adhesion molecule-1 expression of injured human vascular endothelial cells[J]. Zhongguo Lin Chuang Kang Fu, 2006,10(27):54-57
刘萍, 章怡祎, 张静生 . 中药复方对损伤人血管内皮细胞表达细胞间黏附分子的影响[J]. 中国临床康复, 2006,10(27):54-57
[3] Han XH, Zhang N, Liu P . Effects of “Guanxinkang Decoction” on electrophysiological characteristics in isolated papillary muscles in ischemia-reperfusion guinea pigs[J]. Shanghai Zhong Yi Yao Za Zhi, 2009,43(3):58-62
韩向辉, 张娜, 刘萍 . 冠心康对豚鼠缺血再灌注心室乳头肌电生理特性的影响[J]. 上海中医药杂志, 2009,43(3):58-62
[4] Wan TC, Ge ZD, Tampo A, Mio Y, Bienengraeber MW, Tracey WR, Gross GJ, Kwok WM, Auchampach JA . The A3 adenosine receptor agonist CP-532, 903 [N6-(2,5-dichlorobenzyl)-3′-aminoadenosine-5′-N-methylcarboxamide] protects against myocardial ischemia/reperfusion injury via the sarcolemmal ATP-sensitive potassium channel[J]. J Pharmacol Exp Ther, 2008,324(1):234-243
[5] Budas GR, Jovanovic S, Crawford RM, Jovanovic A . Hypoxia-induced preconditioning in adult stimulated cardiomyocytes is mediated by the opening and trafficking of sarcolemmal KATP channels[J]. FASEB J, 2004,18(9):1046-1048
doi: 10.1096/fj.04-1602fje
[6] Li J, Li SP . ATP-sensitive potassium channel and myocardial protection[J]. Xin Xue Guan Kang Fu Yi Xue Za Zhi, 2006,15(5):516-518
李娟, 李曙平 . KATP通道与心肌保护[J]. 心血管康复医学杂志, 2006,15(5):516-518
[7] Yokoshiki H, Sunagawa M, Seki T, Sperelakis N . ATP-sensitive K+ channels in pancreatic, cardiac and vascular smooth muscle cells[J]. Am J Physiol, 1998,274(1):25-37
doi: 10.1152/ajpcell.1998.274.1.C25
[8] Seino S, Miki T . Physiological and pathophysiological roles of ATP-sensitive K+ channels[J]. Prog Biophys Mol Biol, 2003,81(2):133-176
doi: 10.1016/S0079-6107(02)00053-6
[9] Backx PH . Sulfonylurea receptor expression heterogeneity suggests chamber-specific roles for sarcolemmal KATP channels in heart[J]. Circ Res, 2008,103(12):1345-1347
doi: 10.1161/CIRCRESAHA.108.189738
[10] Ljubkovic M, Marinovic J, Fuchs A, Bosnjak ZJ, Bienengraeber M . Targeted expression of Kir6. 2 in mitochondria confers protection against hypoxic stress[J]. J Physiol, 2006,577(1):17-29
doi: 10.1113/jphysiol.2006.118299
[11] Bednarczyc P, Dolowy K, Szewczyk A . New properties of mitochondrial ATP-regulated potassium channels[J]. J Bioenerg Biomembr, 2008,40(4):325-335
doi: 10.1007/s10863-008-9153-y
[12] Auchampach JA, Grover GJ, Gross GJ . Blockade of ischaemic preconditioning in dogs by the ATP dependent potassium channel antagonist sodium 5-hydroxydecanoate[J]. Cadiovasc Res, 1992,26(11):1054-1062
doi: 10.1093/cvr/26.11.1054
[13] Armstrong SC, Liu GS, Downey JM, Ganote CE . Potassium channels and preconditioning of isolated rabbit cardiomytes: effects of glyburide and pinacidil[J]. J Mol Cell Cardiol, 1995,27(8):1765-1774
doi: 10.1016/S0022-2828(95)90986-9
[14] Liu P, Zhang JS, Zheng F, Fu WN, Chen RX . The influence of traditional Chinese medicine on expression level of low-density lipoprotein receptor gene[J]. Zhongguo Zhong Xi Yi Jie He Ji Jiu Za Zhi, 2000,7(3):141-143
刘萍, 张静生, 郑菲, 富伟能, 陈汝兴 . 中药对低密度脂蛋白受体基因表达的影响[J]. 中国中西医结合急救杂志, 2000,7(3):141-143
[15] Yang XX, Zhang JS . Protective effect of Guanxinkang capsule against myocardium injury induced by acute ischemia in rats[J]. Zhong Yi Yao Xue Kan, 2006,24(2):238-241
杨晓霞, 张静生 . 冠心康对实验性大鼠急性肌缺血的保护作用[J]. 中医药学刊, 2006,24(2):238-241
[16] Ren YJ, Tong XY, Wang XL . Expression of ATP-sensitive potassium channels subunit in rat tissues[J]. Zhongguo Yao Li Xue Tong Bao, 2000,16(2):148-150
任亚军, 佟晓永, 王晓良 . ATP敏感钾通道亚单位在大鼠组织中的表达[J]. 中国药理学通报, 2000,16(2):148-150
[1] Li-xin Ma, Yu-yi Wang, Xin-xue Li, Jian-ping Liu . Systematic review on methodology of randomized controlled trials of post-marketing Chinese patent drugs for treatment of type 2 diabetes. Journal of Chinese Integrative Medicine, 2012, 10(3): 279-292.
[2] Zhu Yan-feng, Luo Hai-ming, Deng Zhong-long, Fu De-yu, Yao Wang, Dai Jian. Effects of the Chinese patent medicine, Honghua Injection, on platelet glycoprotein Ⅱb/Ⅲa receptors in patients with acute coronary syndrome: a randomized controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(3): 318-323.
[3] Qi-hong Shi , Jun Xiang, Xu-ying Zhu , Ding-fang Cai. Protective effects of Chinese herbal medicine Naoshuantong on neurovascular unit in rats with cerebral ischemia/reperfusion injury. Journal of Chinese Integrative Medicine, 2012, 10(10): 1135-1139.
[4] Hao Dai, Jun Gu, Ling-zhi Li, Li-mei Yang, Hui Liu, Jian-yu Li. Scutellarin benzyl ester partially secured the ischemic injury by its anti-apoptosis mechanism in cardiomyocytes of neonatal rats. Journal of Chinese Integrative Medicine, 2011, 9(9): 1014-1021.
[5] Susuana Kuek, Wen-jun Wang, Sui-qi Gui. Efficacy of Chinese patent medicine Tian Gui Capsule in patients with polycystic ovary syndrome: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2011, 9(9): 965-972.
[6] Yan Shen, Hong Wei, Chun-lan Jin. Effects of Chinese herbal medicine Xuefu Zhuyu Decoction on angiotensin Ⅱ-induced proliferation and extracellular matrix synthesis of rat cardiac fibroblasts. Journal of Chinese Integrative Medicine, 2011, 9(3): 313-319.
[7] Jun-lin Hu, Gui-lin Ouyang, Da-peng Han, Qing Xia, Yong He, Zheng Huang, Fei Zhu, Song-tao Sun. Effects of Tuina and Chinese patent drug Shuxuetong injection on deep venous thrombosis and functional rehabilitation after total knee arthroplasty. Journal of Chinese Integrative Medicine, 2011, 9(10): 1088-1093.
[8] Ming-wei Huang, Huan Wang, Wen-juan Zhong, Xiao-ying Wu, Hui Chen. Chinese herbal medicine Naoxintong capsule combined with dual antiplatelet therapy in a rat model of coronary microembolization induced by homologous microthrombi. Journal of Chinese Integrative Medicine, 2011, 9(1): 38-48.
[9] Hua Tan, Xiao-ping Huang, Chang-qing Deng. Effects of astragalosides and Panax notoginseng saponins combination on oxidative stress of cerebral ischemic reperfusion injury in mice. Journal of Chinese Integrative Medicine, 2010, 8(5): 448-452.
[10] Da-wu Zhang, Lei Zhang, Jian-gang Liu, Cheng-long Wang, Da-zhuo Shi, Ke-ji Chen. Mechanisms of Chinese herbs combined with ischemic postconditioning in protecting myocardium of rats from ischemia-reperfusion injury. Journal of Chinese Integrative Medicine, 2010, 8(5): 465-471.
[11] Xue-ying Wang, Jie Wang, Ge Yang, Yong-mei Liu. Effects of Xuesaitong Soft Capsule on hemodynamics and cardiocyte apoptosis of rats after myocardial infarction. Journal of Chinese Integrative Medicine, 2010, 8(3): 269-274.
[12] Bin Cheng, Xin Wang, Lei Wang, Kai Cao, Lei Duan, Feng-tao Li. Changes of mitochondrial apoptosis in spinal cord ischemia-reperfusion injury and the effects of Herba Erigerontis Breviscapi Injection preconditioning intervention in rabbits. Journal of Chinese Integrative Medicine, 2009, 7(9): 842-847.
[13] Shao-hua Song, Xiao-yun Shen, Fang Liu, Yi Tang, Zhen-meng Wang, Zhi-ren Fu. Protective effects of astilbin on renal ischemia-reperfusion injury in rats. Journal of Chinese Integrative Medicine, 2009, 7(8): 753-757.
[14] Hong-xing Zhang, Qiong Wang, Li Zhou, Ling-guang Liu, Xuan Yang, Min Yang, Yin-ni Liu, Xuan Li. Effects of scalp acupuncture on acute cerebral ischemia-reperfusion injury in rats. Journal of Chinese Integrative Medicine, 2009, 7(8): 769-774.
[15] Dong-mei Zhang , Ying Qin , Xi-ying Lu , Ai-ming Wu , Ling-qun Zhu , Shuo-ren Wang , Liang-duo Jiang . Effects of tetramethylpyrazine on angiotensin Ⅱ-induced proliferation and type Ⅰ collagen synthesis of rat cardiac fibroblasts. Journal of Chinese Integrative Medicine, 2009, 7(3): 232-236.
Full text



[1] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. An explanation on "guiding principles of clinical research on mild cognitive impairment (protocol)". Journal of Chinese Integrative Medicine, 2008, 6(1): 15-21
[2] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[3] Jun Hu, Jian-ping Liu. Non-invasive physical treatments for chronic/recurrent headache. Journal of Chinese Integrative Medicine, 2008, 6(1): 31
[4] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[7] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[8] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[9] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
[10] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035