Effects Of Different Acupuncture Stimuli On Uterine Microcirculation In Rats With Cold Coagulation And Dysmenorrhea And Intervention Mechanisms

Primary dysmenorrhea(PD)is a commongynecological,functionaldisorder di seasewith nonstructuralchangesinfemale genital mutilation.Itsclinical manif estations are pain in abdomen mainlyoccurred inwomenaftermenstruation,caus ed by manyrisk factors,pathogenesis is not clear recently.According totrad itional chinese medicinesyndrome differentiation,uteruschillis maincause P D.The subject of previous studiesfound thatPD was associated withuterinemic rocirculation disorder,acupuncture SP6can relievepain,improveuterinemicroci rculation,as well confirmedSP6asspecificacupointsto relieve pain.Theexper imentcontinue toselectSP6,using different stimulations to treatcoldsyndrom e ofPDin rats,observe acupunctureefficacyin writhing,uterinemicrocirculatio n and contractive materials,in order toexplorerelevantintervention mechanis m,discusscorrelation of stimulations andefficacy,which correlatedtodeqiand efficacy,to certify deqi impact efficacy.ObjectiveThe objective of the experimentwas to compare effects of treating colds yndrome ofPDmodel rats by acupuncturing SP6 with different stimulations,obs erve changes of writhing,uterinemicrocirculation and contractive materials.The experiment have two stimulations,namely A stimuli(deqi)andB stimuli(no deqi).The results will reveal the mechanism of acupuncture alleviatingp ain,as well as the relationship between stimulations and efficacy,which wi 11 be evidences to explore correlation of deqi and efficacy.Methods64 dioestrum female rats were randomly divided into 4 groups,saline co ntrol group,model control group,A stimuli group,B stimuli group,with 8 rats in each group.Rats of model control group and stimulus groups were fr ozen in-25? freezer for five days with 4 hours per day while given subcut aneous estradiol benzoate injection for ten days with 1 hour after the last injection,intra-peritoneal injection of oxytocin 2u/each.Saline control group were given the same dose of saline for ten days with 1 hour after the last injection,intra-peritoneal injection of saline 2u/each.On the tenth day,stimulus groups were given different quantity of acupuncture stimulus on both sides of "sanyinjiao(SP6)" for 20 minutes.Then observe writhing r esponses and uretine microcirculation respectively.The latency period,the number and score of writhing were recored for 20min in each group.Using XW-B-3 Cold Light microcirculationtesterobservethechangesofuterine microcircu lationeachgroupsat different times of 5min,10min,20min,30min,40min,50min,60 min.After recording the writhing responses,materials including OTR,Ca2+ an d MLCK were measured respectively by different methods such as REALTIME-PCR,ELISA,and MethylThymolBlue Method.Results1 Effect of different stimulations of acupuncture on the writhing responseCompared with saline control group,the latency period of writhing in mo del control group was shortend significantly,the number and score of writhi ng was increased significantly(P<0.01).In comparison with model group,the latency period of writhing was lengthened and the number and score of writh ing was decreased in A stimuli group(P<0.05,P<0.01).The score of writhin g in B stimuli group was decreased(P<0.05).The latency of writhing in A st imuli group was longer than in B stimuli group(P<0.05).2 Effect of different stimulations on uterus microcirculation of female rat s at different timesModel control group comparing to saline control group:diameters of mic ro-vessel at all time-points,diameters of capillary(except40min and 60min)w ere significantly contracted(P<0.01),numbers of capillary were reduced sig nificantly(P<0.01),clarity of micro-vessel was worse,color darker,blood f low slower or stagnation(P<0.Olor P<0.05).Stimulus groups comparing to mod el control group:about A stimuli group,diameters of micro-vessel at all t ime-points,diameters of capillary(only5min,10min,20min and 30min)were sig nificantly expanded(P<0.01orP<0.05),clarity of micro-vessel(except20min)was better,color(only 10min,20min,40min and50min)redder,blood flow at all t ime-points was speed up significantly(P<0.01or P<0.05).About B stimuli grou p,diameters of micro-vessel were expanded only at 30min(P<0.01),clarity of micro-vessel(only 50min and60min)was better(P<0.05).A stimuli group comp aring to B stimuli group:diameters of micro-vessel at all time-points,dia meters of capillary(only5min,10min,20min and30min)were significantly expa nded(P<0.01orP<0.05),clarity of micro-vessel(only30minand 40min),color(onl y 5min,20min and40min),blood flow(except50min and60min)were obviously impr oved(P<0.01orP<0.05).3 Effect of different stimulations of acupuncture on materials in uterine i ncluding OTR,MLCK and Ca2+ of female ratsCompared with each other,the expression of OTR and the density of Ca2+in four groups were no difference.Compared with model group,the content of MLCK increased significantly(P<0.05),Compared with model group,the conte nt of MLCK in B stimuli group was decreased(P<0.05).Conclusions1 AcupunctureSP6 can relieve pain in cold syndrome dysmenorrhea of female r ats,different stimulations made different efficacy.A stimuli group("deqi"group)relieve pain obviously,confirmed that stronger stimuli got better efficacy in the treatment of primary dysmenorrhea.2 A stimuli group(“deqi”group)can improve uterine microcirculation,reliev e pain obviously.B stimuli group(no "deqi" group)can imfact uterine microc irculation softly.3 Compared to B stimuli group(no "deqi" group),A stimuli group("deqi" gro up)got better efficacy,which showed that A stimuli group("deqi" group)was in better“deqi”state,B stimuli group(no“deqi”group)in worse "deqi"state.4 It showed no obvious changes and rules in the expression of OTR,the densi ty of Ca2+.Compared with model group,the content of MLCK changed obviously i n B stimuli group(no "deqi" group).The results may be caused by two reasons.(1)With different stimulations,there were differences in the role of regula ting aspects.(2)Cold stress,which disorder inner balance to cause patholog ical changes such as receptor inactivation,inhibition of kinase activity,blocked ion channel activity,which need further explore.In summary,this study showed that different stimulations made diferent efficacy and regulated by different mechanism.A stimuli group("deqi" gro up)showed better analgesic effect in the better“deqi”state,illustrated the importance of "deqi" in the acupuncture efficacy.