[Effect of different acupuncture and moxibustion methods on uterine contraction and microcirculation in cold congealing dysmenorrhea rats].

To observe and compare the effects of different acupuncture and moxibustionmethods at "San-yinjiao" (SP6) on uterine contraction and microcirculation in cold congealing dysmenorrhea rats, so as to explore its mechanism underlying treatment of dysmenorrhea.A total of 140 female SD rats were randomly divided into normal control, mo-del, perpendicular needling, transverse needling and moxibustion groups, with 28 rats in each group. The cold congealing dysme-norrhea rat model was prepared by exposure in a freezer (25 ℃) for 4 h, once daily for 5 days, and subcutaneous injection of estradiol benzoate (once daily for 10 days) and intra-abdominal injection of oxytocin (once). For rats in three intervention groups, acupuncture needles were inserted into bilateral SP6 perpendicularly or transversely to a depth of 4~5 mm and retained for 20 min, or moxibustion was applied to SP6 for 20 min. The uterine contraction degree and the uterine microcirculation were recorded. The expression levels of transient receptor potential vanilloid 1 (TRPV1) and heat shock protein 70 (HSP70) in local tissues of SP6 area were detected by immunohistochemistry. The expression levels of μopioid receptor and endothelin 1 (ET1) mRNA in the uterus were assessed by quantitative real time-PCR.After modeling and compared with the normal control group, the number and peak-to-peak values of uterine contraction waves, and uterine motility were significantly increased (P<0.01,P<0.05), while the speed of blood flow in the microvessels was slowed down (P<0.01), diameters of the uterine microvessels and capillaries (cap) shrank obviously (P<0.01) in the model group. After the intervention, all indexes of uterine contraction and microcirculation were improved in three intervention groups (P<0.01, P<0.05), while transverse needling and moxibustion showed better effects compared to perpendicular needling (P<0.05, P<0.01). The expression of TRPV1 and HSP70 in SP 6 area had no significant changes (P>0.05), while the uterine μopioid receptor mRNA expression decreased (P<0.01), and ET1 mRNA expression increased (P<0.01) in the model group relevant to the normal control group. Following the intervention, the expression levels of TRPV1 and HSP70 in SP6 area were increased (P<0.05, P<0.01), and expression levels of uterine μopioid receptor mRNA increased (P<0.05, P<0.01) and uterine ET1 mRNA decreased (P<0.05, P<0.01) in the three intervention groups. The effect of moxibustion was considerably better than those of two acupuncture groups in up-regulating TRPV1 expression (P<0.05). Both transverse needling and moxibustion showed better effects of down-regulating uterine ET1 mRNA expression than perpendicular needling (P<0.01).Transverse needling and moxibustion at SP6 have a better effect of relieving ute-rine contraction and improving uterine microcirculation than perpendicular needling, which may be related to their effects in up-re-gulating the expression of TRPV1 and HSP70 in SP6 area, thereby modulating the mRNA expression of μ opioid receptor and ET1 in uterine tissue.目的：比较针刺、艾灸“三阴交”穴对寒凝证类痛经大鼠子宫收缩程度及微循环的影响, 探讨不同刺灸法改善痛经的作用机制。方法：雌性SD大鼠随机分为空白组、模型组、直刺组、平刺组、艾灸组, 每组28只。采用全身冷冻加雌二醇注射法制备寒凝证类痛经模型。3个干预组分别直刺、平刺、艾灸“三阴交”穴20 min。观察记录各组大鼠子宫收缩程度及微循环状况。免疫组织化学法检测“三阴交”穴区香草酸瞬时受体亚型1（TRPV1）、热休克蛋白70（HSP70）的表达, 荧光定量PCR法检测子宫组织μ阿片受体及内皮素（ET）1 mRNA表达。结果：与空白组比较, 模型组宫缩增强(P<0.01), 子宫微血流状态评分升高、毛细血管与微血管管径缩小(P<0.01)。与模型组比较, 3个干预组宫缩减弱、子宫微血流状态评分降低、毛细血管与微血管管径增大(P<0.01, P<0.05)。平刺与艾灸组均较直刺组宫缩及微循环改善(P<0.05, P<0.01)。与空白组比较, 模型组穴区TRPV1、HSP70的表达差异无统计学意义(P>0.05), 子宫μ阿片受体mRNA表达降低(P<0.01), ET1 mRNA表达升高(P<0.01)。与模型组比较, 3个干预组穴区TRPV1、HSP70表达均增强(P<0.05, P<0.01), 子宫μ阿片受体mRNA表达升高(P<0.05, P<0.01), ET1 mRNA表达降低(P<0.05, P<0.01)。艾灸组穴区TRPV1表达高于两个针刺组(P<0.05), 平刺组与艾灸组ET1 mRNA表达低于直刺组(P<0.01)。结论：3种刺灸法均不同程度改善了痛经大鼠的宫缩与微循环, 平刺和艾灸优于直刺, 可能与其激活穴区TRPV1、HSP70, 进而调节子宫μ阿片受体、ET1基因表达有关。.