| Purpose: To observe the effects of KATP gene Kir6.1,Kir6.2,compound protein SUR2 A, SUR2 B, and protein kinase PKA,PKC-β2 m RNA expression of myocardial ischemia mice through acupuncturing different acupoints with the method of RT-PCR. To investigate the mechanism of acupuncture at Neiguan from the point of protein kinase gene expression difference between that in myocardial cells KATP and information transmission system. To reveal the specificity of acupoints and provid a scientific basis for the treatment of myocardial ischemia through acupuncture.Material and method:52 SPF ASIC3-/- mice were regarded as the research object, 8 among them were randomly selected according to random number table as the control group. The remaining 44 mice were injected subcutaneously multi-point isoproterenol hydrochloride(20mg·kg-1·d-1),while mice in control group were injected with an equal amount of saline, and electro cardiograms were recorded before and after modeling in mice. Excluding 4 dead mice,remaining 40 successful model mice were randomly divided into five groups of 8: model group, Neiguan group, Lieque group, Zusanli group and non-acupoint group which were applied of Electro-acupuncture. Huatuo sterile acupuncture needles(φ0.19×13mm)were used to pierceinto the skin for 2mm, and connected G6805-AⅡelectroacupuncture device, subjected to density wave at the frequency of 4-20 Hz with the intensity of 2m A. Retaining needles for 20 min a day, and lasted for 7 days. The Electrocardiograph in each group were recorded after the end of treatment. None of acupuncture was used in mice of control group and model group. However, mice in both of these two groups were crawled and fixed at the same way every day. The mice were drawn materials at animal experimental research center of Liaoning University of Traditional Chinese Medicine which were killed by breaking off the ridge. Under sterile conditions, hearts were quickly removed and the left ventricle were separated.Put the left ventricle tissues into sterilized and numbered EP tube, with 1ml the trizol reagent added and were rapidly frozen with liquid nitrogen, then they were immediately placed in-80℃ ultra-low temperature freezer. Using to detect the changes of expression levels about myocardial adenosine triphosphate-sensitive potassium ion channel-related geneskir6.1, kir6.2, SUR2 A, SUR2 B and myocardial cell transduction system gene related protein kinase PKA, PKC-β2 and the reference gene of β-action were detected by using the real-time quantitative polymerase chain reaction(RT-PCR)for statistical analysis.Results: 1.Compared with control group, the relative expression of target genes in model group, Neiguan group, Lieque group, Zusanli group, non-acupoint group are significant increased(P<0.05,P<0.01).2.Compared with model group, the relative expression of target genes in Neiguan group, Lieque group and Zusanli group are significantly lower(P<0.05,P<0.01), differences in nonacupoint group have not statistically significance(P>0.05).3.Compared with Neiguan group, the relative expression of target genes in Lieque group and Zusanli group are significantly increased(P<0.05,P<0.01).4.Compared with Lieque group, the relative expression of target genes in non-acupoint group are significantly increased(P<0.05,P<0.01), while differences in Zusanli group do not have statistically significance(P>0.05).5.Compared with the Zusanli group, therelative expression of target genes in non-acupoint group are significantly increased(P<0.05,P<0.01).Conclusion: 1.Electroacupuncture at Neiguan, Lieque and Zusanli are effective for the treatment of myocardial ischemia. 2.Compared with Lieque and Zusanli, the therapeutic effects of Neiguan for myocardial ischemia are more significant. 3.Electroacupuncture at Neiguan has meridian specificity. 4.Meridian specificity is not affected by pain interference. |
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