Effects Of Different Acupuncture Stimuli On Uterine Contraction Intensity, Microcirculation And Expression Of Related Molecular Receptors In Rats With Cold Coagulation And Dysmenorrhea

Objective:To evaluate the effects of acupuncture stimulation with different amount at Sanyinjiao(SP6)on uterine contraction,uterus microcirculation and uterine endocrine related molecular receptor in rats with dysmenorrhea of cold coagulation syndrome,analyze the correlation between this three aspects under dysmenorrhea and to explore whether there is direct relativity between "De qi" and needle stimulus intensity.Methods:96 female Sprague Dawley rats were screened by vaginal smear estrous cycle test and randomly divided into four groups:saline control group,cold coagulation syndrome types of dysmenorrhea model group(model group),"A" stimulus group(with strong stimulus)and"B" stimulus group(with weak stimulus).Model group and treatment groups were designed to use the general freezing method combined with the estradiol benzoate injection molding.After the preparation of the model,96 rats were divided into 3 groups and each group has 32 rats.Each group was divided into four smaller groups.The first group of 32 rats were used for uterine contraction experiment with using BL-420F biological experimental system recorder,to record the uterine contraction waves and related numerical.Second group of 32 rats were used for uterine microcirculation experiment which using XW-B-3 cold light source microcirculatory microscopic examination instrument to observe the diameter and blood flow parameters in a fixed visual field at 5,10,20min respectively.The third set of experiments using RT-PCR detected in the rat uterus PGE2,PGF2a receptor mRNA and OTR mRNA'expression.Result:1.Intensity of Uterine Contraction:Compared with the saline control group,the uterine contraction wave number and uterine activity of the model control group were significantly increased.Meantime,compared with the model control group,the uterine contraction wave number and uterine activity were significantly reduced by"A" stimulus applied to SP6 in rats with dysmenorrhea of the cold-coagulation syndrome.2.Uterine Microcirculation:Compared with the saline control group,the uterine microvascular and capillary diameter of the model control group were significantly decreased,and markedly increased by "A" stimulus applied to SP6 compared with the model control group.Compared with "B" stimulus group,"A" stimulus group significantly expanded on 5min,20 min(P<0.05)and 10min(P<0.01).Compared with the saline control group,blood flow was slower in the model group,and significantly quicker by "A" stimulus during acupuncture treatment.3.Expression of PGE2,PGF2? Receptor and OTR mRNA in Rat Uterus Tissue:Compared with the saline group,PGF2?and PGE2 mRNA expressions of model group were increased.(P<0.01)(P<0.05).Compared with the model group,"A" stimulus group,both PGF2aand PGE2 mRNA were decreased expression(P<0.05)(P<0.01),the expression of PGF2aand PGE2 receptor mRNA of "B" stimulus group were reduced,but the difference was not statistically significant(P>0.05).In OTR mRNA ecpression test,the difference of all groups without statistically significant(P>0.05).4.Uterine contraction and Uterus Microcirculation Correlation:The relationship of uterine microvessel capillary diameter and the contraction of uterus showed negative correlation.5.Correlation of PGF2? receptor mRNA Expression,Uterine contraction and Uterus Microcirculation:PGF2a receptor mRNA expression and uterine contraction wave had positive correlation;meantime,the relationship between PGF2? receptor mRNA expression and uterine microcirculation diameter showed negative correlationConclusion:1.Under cold coagulation syndrome dysmenorrhea environment,the uterine contraction of model rats had increased,uterus microcirculation diameter was decreased,blood flow was slower and increased PGF2aand PGE2 receptor mRNA expression,it showed there have a closely correlation these three aspects.2.Acupuncture applied at Sanyinjiao(SP6)can help to improve PGF2aand PGE2 receptor mRNA expression,uterine contraction and uterus microcirculation of cold coagulation syndrome dysmenorrhea rats.3."A" stimulus leads to better outcomes on reducing PGF2aand PGE2 receptor mRNA expression,uterine contraction and uterus microcirculation of cold coagulation syndrome dysmenorrhea rats.4.Although,compare with "A" stimulus group,most of the treatment effects of "B"stimulus group(the weak stimulus)without satisfied the statistical significant,the weak stimulus also can help to reducing PGF2?and PGE2 receptor mRNA expression,uterine contraction and uterus microcirculation of cold coagulation syndrome dysmenorrhea rats.5.Our finding highlights using thick needles,deep insertion with hand-manipulation to cause appropriate intensity of stimuli to achieve "De qi" state in acupuncture treatment of dysmenorrhea.