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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 E-mail:liuping23@sina.com

Objective

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.
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 ℃. 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.
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 KATP 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.

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."

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