Targeting Regulation Of Chloride Channel Related Genes In Cardiomyocytes By Acupuncture At Neiguan

Purpose:This experiment is based on the model of myocardial ischemia(MI)Acid-sensing ion channels 3 gene konckout(ASIC3-/-)mice had been made by injecting isoproterenol(ISO),and is to observe the effects of acupuncture at different meridians on the expression of Cl Ca1、Cl C2、Cl C3 and CFTR in the schloride channel with cardiomyocytes of myocardial ischemia gene knockout(ASIC3-/-)mice.Meanwhile,through the comparison with our previous experiment in the effect of acupuncture that the effect of acupuncture at different acupuncture points on the expression of chloride channel related genes in rats,to further explore the targeted role of acupuncture at Neiguan acupuncture on chloride ion channel related genes in cardiomyocytes,and reveal the mechanism of specificity of meridian effect.Material and method:According to a table of random number table method,43 SPF grade acid-sensitive ion channel 3 gene knockout(ASIC3-/-)mice were divided into 5groups: Blank group,Model group,Neiguan group,Lieque group and neither meridians nor points group.Except for blank group,the remaining mice were given subcutaneous injections of ISO(20 mg/kg)twice(interval of 24 hours)for preparation of myocardial ischemia model.After confirmation of the success of modeling according to modeling before and after ECG changes,"Hua Tuo" brand acupuncture needles(0.19 mm × 13mm)were applied and inserted into each corresponding point 1-2mm under the skin,along the 30 °oblique thorn.An electro-acupuncture(EA)device was connected and the mice were subjected to a density wave(2 Hz).Rats were stimulated for 20 min once a day,seven times consecutively.The genes’ expression of Cl Ca1,Cl C2,Cl C3 and CFTR were analyzed by Real-time PCR.Results:1.Compared with modeling,Neiguan group the T-wave voltage of mices’ ECGsignificantly increased(P<0.05).Compared with blank group,the T-wave voltage of model group、Neiguan group、Leique group and neither meridians nor points group significantly increased,respectively(P < 0.05).Compared with the model group,the T-wave voltage of Neiguan group significantly increased(P<0.05)and T-wave voltage of mice’ ECG did not change significantly in the Leique、 neither meridians nor points group(P>0.05).The effects of Neiguan group were higher than Lieque、neither meridians nor points group(P<0.05);there are no significant difference between the group of Lieque and neither meridians nor points group(P>0.05).2.Compared with the blank group,the genes expression of Cl C2、Cl C3 、Cl Ca1 and CFTR were significantly increased in model group 、 Neiguan group 、 Leique group and neither meridians nor points group(P<0.05).Compared with the model group,except for the neither meridians nor points group which has no significant difference in the expression of Cl C3(P>0.05),the genes expression of Cl C2、Cl C3、Cl Ca1 and CFTR with Neiguan 、Leique and neither meridians nor points groups were decreased(P<0.05).Compared with the Neiguan group,the genes expression of Leique and neither meridians nor points groups increased(P<0.05);Compared with the Leique group,the expression of Cl C2、Cl C3 and CFTR were significantly increased(P <0.05),and there was no significant difference in the expression of Cl Ca1 with neither meridians nor points groups(P> 0.05).3.At the end of acupuncture treatment,after comparing the gene knockout(ASIC3-/-)mice with the same experimental rats in the gene expression of each group.The results showed that the differences between CFTR、Cl C3、Cl Cal and CFTR gene expression groups were not significantly different from those of rats,and the other expression trends were the same.Conclusion:1.Acupuncture Neiguan acupuncture point can effectively regulate the abnormal changes of T wave voltage in ECG of mice with myocardial ischemia gene knockout(ASIC3-/-),while reducing the increase of Cl Ca1,Cl C2,Cl C3 and CFTR gene expression in cardiomyocytes of myocardial ischemia gene knockout(ASIC3-/-)mice,and the same asthe rats` experimental results.2.Neiguan of pericardium meridian and Lieque of Lung meridian can improve the genes expression of chloride channel,and the effect of Neiguan is better than Leique.Indicating that acupoints of acupuncturing acupoints have a targeted effect on chloride ion channel responses in cardiomyocytes.