Study Of The Effects Of Various Quantities Of Acupuncture Stimulation On The Body Temperature And Antinociceptive Substances In The Rat With Dysmenorrhea Of Cold Stagnation Type

Objective:In the present experiment Estradiol Benzoate and Oxytocin were selected to make experimental primary dysmenorrheal model. Rats were put into freezer to make cold syndrome model. Two methods were combined to establish experimental cold syndrome of primary dysmenorrheal model. Sanyinjiao(SP6) was selected for the experiment. Different amount of acupuncture were treated in A Stimuli Group and B Stimuli group. Rats in A stimuli group were treated by thicker needles,deeper insertion and manipulation Rats in B stimuli group were treated by shallow insertion of thin needle with no manipulation. Intensity of uterine contraction Temperature of Sanyinjiao(SP6) and Xuehai(SP10) were recorded . MOR and KOR mRNA in uterine and spinal cord and NMDA receptor in hippocampus were detected by Realtime-PCR. All these data were analyzed to estimate analgesia of different acupuncture methods in cold syndrome of primary dysmenorrhea rats.The result may be used to investigate objective evaluation criterion and and the biological mechanism of "deqi".Methods:96 female rats in anestrus were randomly divided into 4 groups, which included saline control group,cold syndrome of primary dysmenorrheal model control group(model control group), A Stimuli Group and B Stimuli Group. Saline control group rats were injected 0.9%Nacl.All other rats were injected Estradiol Benzoate.From the first day to the fifth day rats (except saline control group) were put into-25 ℃ freezer to make cold syndrome of primary dysmenorrheal model rats. Rats in A stimuli group were treated by Φ0.25×40mm needle insertion on Sanyinjiao(SP6) for 4-5mm with 30-second needle manipulation of even reinforcement and reduction at the beginning and 10 minutes after the insertion. Rats in B stimuli group (shallow insertion of thin needle with no manipulation) were treated byΦ0.18×13mm needle insertion on Sanyinjiao (SP6) for 1-2mm with no needle manipulation. BL-420F logger of experimental system was applied to record times and intensity of uterine contraction in 20minutes.Temperature changes of 2-side Sanyinjiao (SP6), Xuehai (SP10) in 60 minutes were recorded. MOR and KOR in uterine and spinal cord were detected by RealTime-PCR. NMDA receptor in hippocampus was detected by RealTime-PCR.Result:1 Effet of Different acupuncture amount on Degree of Uterine Contraction of cold stagnation dysmenorrheal rats:Model control group comparing to the saline control group: writhing latency significantly decreased(P<0.01), writhing number and writhing number significantly increased(P<0.01). A Stimuli Group comparing to the cold syndrome of primary dysmenorrheal model group: writhing latency significantly increased(P<0.01), writhing number and writhing number significantly decreased(P<0.01). B Stimuli Group comparing to A Stimuli Group writhing latency significantly decreased(P<0.01), writhing number and writhing number significantly increased(P<0.01).2 Effects of acupuncture with different acupuncture stimuli on temperatures of the Sanyinjiao (SP6) and Xuehai (SP10) of cold syndrome of primary dysmenorrheal model group.The cold syndrome of primary dysmenorrheal model control group showed left-right asymmetric temperature of Sanyinjiao (SP6) point. The asymmetric temperature of left-right Sanyinjiao (SP6) point can be accommodated in A Stimuli Group to new balance state after 20-30 minutes of the end of needling methods. No significant change was detected in B Stimuli Group.3 Effet of Different acupuncture amount on the expression of μ- and k- opioid receptors in uterus and spinal cordCompared with cold syndrome of primary dysmenorrheal model : the expression of μ -opioid receptor mRNA in uterus tissue was increased:A Stimuli Group(P<0.01)and B Stimuli Group(P<0.05).No significant change of κ- opioid receptor in uterus was detected.No significant change of μ- and κ- opioid receptors in spinal cord were detected.4 Effet of Different acupuncture amount on the expression of NMDA receptor in hippocampus of cold stagnation dysmenorrheal ratsNo significant change of NMD A receptor in hippocampus was detected.Conclusion:1 A Stimuli Group can reduce spasmodic contraction of the smooth muscle of uterus significantly, which is significantly better than B Stimuli Group.2 Bigger stimuli(A Stimuli Group) may significantly accommodate the asymmetric temperature of left-right Sanyinjiao (SP6) point to new balance. The time of course is 20-40 minutes after of the end of needling methods. Smaller stimuli(B Stimuli Group) showed no significantly change. A Stimuli Group showed better analgesia effect and accommodate the asymmetric temperature of left-right Sanyinjiao (SP6) point. Based on the traditional Chinese medicine theory "After deqi(Arrival of qi) , the acupuncture can be effective", we confirmed that A Stimuli Group was in better"deqi" state.3 In A Stimuli Group the expression of μ -opioid receptor mRNA in uterus tissue was significantly increased, which proved that the analgesia mechanism involves regulation of the expression of opioid peptides of the target organs. In the present experiment the expression of μ- and κ- opioid receptors in uterus and spinal cord and NMDA receptor in hippocampus showed no significantly change, which maybe related to the influence of cold stress to the sympathetic nervous system.