| Objective:To explore meridian-acupoints specialty and acupoints curative difference,uterus contraction difference,skin temperature changes on acupoints area,κ opioid receptor and μ opioid receptor in central and peripheral tissues were investigated by needling SP6 and CV4 on cold congealing and dysmenorrhea rats.Methods:Method 1:Thirty two female Sprague Dawley(SD)rats were randomly-divided into four groups by using a random number table.The four groups included Saline Control Group(n=8),cold congealing and dysmenorrhea Model Group(n=8),Group SP6(Sanyinjiao)(n=8),Group CV4(Guanyuan)(n=8).Except Saline Control Group,other groups were subcutaneously injected Estradiol Benzoate for 0.5 mg/rat on the 1st and 10th day,and 0.2 mg/rat from the 2nd to the 9th day.Then the rats were placed in a freezer for 4 hours at the first 5 days with temperature maintained at-25 ℃ and the freezer was opened for 5 seconds during 2 hours interval.One hour later after the injection of Estradiol Benzoate on the 10th day,0.7 ml/100 g Urethane was given by intramuscular injection for anesthesia.Then abdomen surgery was operated,and BL-420F was used to record the uterus contraction.Method 2:Thirty two female Sprague Dawley(SD)rats were randomly and equally divided into four groups by using a random number table.The four groups included Saline Control Group,Model Group,Group SP6,and Group CV4.Infrared thermal imaging was applied to investigate the skin temperature of SP6,SP10,line between SP6 and SP10,CV4 and the uterus area at different time points,such as baseline,instant,5 min,10 min,20 min,30 min,40 min,50 min,60 min after needling.Method 3:Thirty two female Sprague Dawley(SD)rats were randomly and equally divided into four groups by using a random number table.The four groups included Saline Control Group,Model Group,Group SP6,and Group CV4.The rats were killed when acupuncture finished,the spinal cord and uterus were removed soon afterward.RealTime-PCR were used to test the amount of κ opioid receptor and μ opioid receptor in the spinal cord and the uterus.Results:Result 1:Compared with the saline control group,the number of the contraction waves,peak to peak value,and degree of contraction in Model group increased significantly(p<0.01,p<0.05,p<0.01);Compared with Saline control group,degree of contraction in both Group SP6 and Group CV4 increased prominently(p<0.01,p<0.05);Compared with Model Group,the number of uterus contraction waves and degree of contraction in both Group SP6 and Group CV4 decreased significantly(p<0.05,p<0.05).Result 2:(1)Comparison of skin temperature changes on left SP6 at different time intervals:Compared with Saline Control Group,Group SP6 and Group CV4 showed that skin temperature changes on left SP6 decreased significantly at instant needling-5 min interval(p<0.01).Compared with Model group,Group SP6 showed that skin temperature changes on left SP6 decreased significantly at instant needling-5 min interval(p<0.05).(2)Comparison of skin temperature changes on right SP6 at different time intervals:Compared with Saline Control Group,skin temperature changes on right SP6 in Group SP6 increased significantlyy at 5 min-10 min interval(p<0.01),of that in Model group decreased at 10 min-20 min interval(p<0.05)and of that in Group CV4 decreased at 10 min-20 min interval(p<0.01).Compared with Model group,Group SP6 showed that skin temperature changes on right SP6 increased significantly at 5 min-10 min,20 min-30 min intervals(p<0.01).Compared with Group CV4,Group SP6 showed that skin temperature changes on right SP6 increased significantly at 5 min-10 min,20 min-30 min intervals(p<0.01).(3)Comparison of skin temperature changes on left SP10 at different time intervals:There was no significant difference at different time intervals among Saline Control Group,Model Group,Group SP6 and Group CV4 as p>0.05.(4)Comparison of skin temperature changes on right SP10 at different time intervals:Compared with Saline Control Group,Model group decreased prominently at 30 min-40 min interval(p<0.05).Group SP6 decreased at instant needling-5 min interval(p<0.01)and increased at 5 min-10 min interval(p<0.01).Group CV4 decreased at instant needling-5min interval(p<0.05).Compared with Model group,Group SP6 decreased prominently at instant needling-5 min interval(p<0.05)and increased at 5 min-10 min,30 min-40 min intervals(p<0.01).Group SP6 decreased prominently at instant needling-5 min interval(p<0.05)and increased at 5 min-10 min interval(p<0.01)when compared to that of Group CV4.(5)Comparison of skin temperature changes on left line between SP6 and SP10 at different time intervals:There was no significant difference at different time intervals among Saline Control Group,Model Group,Group SP6 and Group CV4 as p>0.05.(6)Comparison of skin temperature changes on right line between SP6 and SP10 at different time intervals:Compared with Saline control group,Model group decreased at 10 min-20 min interval(p<0.05):Group SP6 decreased prominently at instant-5 min interval(p<0.01),increased at 5 min-10 min interval(p<0.01),and decreased at 10 min-20 min interval(p<0.05),Group CV4 decreased at 10 min-20 min interval(p<0.01);Compared with Model group,Group SP6 decreased significantly at instant-5 min interval(p<0.05)and increased at 5 min-10 min interval(p<0.01);Compared with Group CV4,Group SP6 increased at 5 min-10 min interval(p<0.01).(7)Comparison of skin temperature changes on CV4 at different time intervals:There was no significant difference at different time intervals among Saline Control Group,Model Group,Group SP6 and Group CV4 as p>0.05.(8)Comparison of skin temperature changes on uterus area at different time intervals:There was no significant difference at different time intervals among Saline Control Group,Model Group,Group SP6 and Group CV4 as p>0.05.Result 3:(1)κ opioid receptor and μ opioid receptor in the spinal cord:There were no significant difference among groups.(2)κ opioid receptor in the uterus:There were no significant difference among groups.(3)μ opioid receptor in the uterus:Compared with Saline control group,u opioid receptor increased significantly in Group CV4(p<0.05);Compared with Model group,μ opioid receptor increased significantly in both Group SP6 and Group CV4(p<0.05,p<0.01).Conclusion:(1)Acupuncture SP6 and CV4 can be used to treat dysmenorrhea by alleviating uterus contraction.(2)Acupuncture SP6 and CV4 can induce analgesia effect by regulating endogenous opioid receptor in peripheral tissues.(3)Skin temperature changes of SP6 and SP10 can be regulated by needling both SP6 and CV4.Skin temperature changes on right SP6,SP10,line between SP6 and SP10 increased in Group SP6.While there was no significant difference occurring in Group CV4.This study shows that there are dynamic temperature changes along meridian after needling,which not only can be used to evaluate De-qi but also provide evidence for the scientificity of specificity of acupoint.This study confirmed the curative effect of SP6 and CV4 in treating primary dysmenorrhea.Skin temperature changes along meridians after needling illustrated the existence of meridian and meridian-acupointsspecialty,which provided experimental evidence for infrared thermal imaging application on De-qi. |
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