Song Yue, Ma Liang-Xiao, Gan Ying-Ying, Wang Jun-Xiang, Mu Jie-Dan, Guo Meng-Wei, Ren Xiao-Xuan, Yu Wen-Yan, Tian Yuan, Qian Xu, Sun Tian-Yi
School of Acupuncture-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China.
Zhen Ci Yan Jiu. 2020 Nov 25;45(11):895-901. doi: 10.13702/j.1000-0607.200184.
To observe the effect of perpendicular and subcutaneous transverse needling at "Sanyinjiao" (SP6) on visceral pain behavior, arginine vasopressin (AVP) content in the serum, uterine tissues, spinal cord and hypothalamus and expression of AVP receptors AVPR1A and AVPR1B in the uterine tissues, spinal cord and hypothalamus in cold-stasis (stasis due to pathogenic cold) type dysmenorrhea rats, so as to explore their mechanisms underlying pain relief.
Forty female SD rats were randomly divided into blank control, model, perpendicular needling and transverse needling groups, with 10 rats in each group. The cold-stasis dysmenorrhea rat model was established 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 injection (once). For rats of the two acupuncture groups, acupuncture needles were inserted into the bilateral SP6 perpendicularly or transversely to a depth of about 4-5 mm, and retained for 20 min. The abdominal pain behavior was assessed by recording the writhing latency and scaling the rats' writhing reactions after modeling. The contents of AVP in the serum, uterus, spinal cord and hypothalamus tissues were assayed using ELISA and the expression of AVPR1A and AVPR1B in the uterus, spinal cord and hypothalamus was measured by using Western blot and quantitative real time-PCR, respectively.
After mode-ling and compared with the blank control group, the writhing latency was significantly shortened (<0.05), and the writhing score in the first 20 min was significantly increased (<0.01) in the model group. After the intervention, the writhing latency was significantly prolonged (<0.01), and the writhing scores in 20 min were significantly decreased (<0.01) in the two needling groups. The AVP contents were obviously increased in the serum and uterine tissue (<0.05, <0.01) but decreased appa-rently in the spinal cord and hypothalamus tissues (<0.01, <0.05), and the expression levels of AVPR1A or AVPR1B protein and mRNA were markedly increased in the uterine tissues (<0.01, <0.05), and significantly decreased in the spinal cord and hypotha-lamus in the model group relevant to the control group (<0.05, <0.01). Following the intervention, The AVP content in the serum of the perpendicular needling group (<0.05) and that in the uterus of the two needling groups were significantly decreased (<0.01), as well as that in the hypothalamus was obviously increased in the two needling groups (<0.05). The expression levels of AVPR1A protein and mRNA in the uterus were significantly down-regulated in the two needling groups (<0.01, <0.05) and AVPR1B protein in the hypothalamus of the perpendicular needling group was up-regulated (<0.05). Moreover, no significant differences were found between the two needling groups in regulating the related indexes mentioned above (>0.05).
Both perpendicular and subcutaneous transverse needling at SP6 have an immediate analgesic effect in cold-stasis type dysmenorrhea rats, which may be related to their effects in regulating AVP levels and its receptor expression in the uterine and hypothalamus.
观察三阴交穴直刺与皮下横刺对寒凝血瘀型痛经大鼠内脏痛行为、血清、子宫组织、脊髓及下丘脑精氨酸加压素(AVP)含量,以及子宫组织、脊髓和下丘脑AVP受体AVPR1A和AVPR1B表达的影响,以探讨其止痛机制。
将40只雌性SD大鼠随机分为空白对照组、模型组、直刺组和横刺组,每组10只。采用在冷冻箱(-25℃)中暴露4 h,每日1次,共5 d,皮下注射苯甲酸雌二醇(每日1次,共10 d)及腹腔注射缩宫素注射液(1次)的方法建立寒凝血瘀型痛经大鼠模型。对两组针刺组大鼠,分别将针刺垂直或横向刺入双侧三阴交穴,深度约4~5 mm,留针20 min。通过记录造模后大鼠扭体潜伏期及对大鼠扭体反应进行评分来评估腹痛行为。采用ELISA法检测血清、子宫、脊髓及下丘脑组织中AVP的含量,分别用Western blot法和实时定量PCR法检测子宫、脊髓和下丘脑中AVPR1A和AVPR1B的表达。
造模后,与空白对照组比较,模型组大鼠扭体潜伏期显著缩短(<0.05),前20 min扭体评分显著升高(<0.01)。干预后,两组针刺组大鼠扭体潜伏期显著延长(<0.01),20 min内扭体评分显著降低(<0.01)。模型组血清和子宫组织中AVP含量明显升高(<0.05,<0.01),脊髓和下丘脑组织中AVP含量明显降低(<0.01,<0.05),子宫组织中AVPR1A或AVPR1B蛋白及mRNA表达水平明显升高(<0.01,<0.05),与对照组比较,模型组脊髓和下丘脑组织中AVPR1A或AVPR1B蛋白及mRNA表达水平显著降低(<0.05,<0.01)。干预后,直刺组血清中AVP含量(<0.05)及两组针刺组子宫中AVP含量均显著降低(<0.01),两组针刺组下丘脑组织中AVP含量明显升高(<0.05)。两组针刺组子宫中AVPR1A蛋白及mRNA表达水平均显著下调(<0.01,<0.05),直刺组下丘脑组织中AVPR1B蛋白表达上调(<0.05)。此外,两组针刺组在调节上述相关指标方面差异无统计学意义(>0.05)。
三阴交穴直刺与皮下横刺对寒凝血瘀型痛经大鼠均有即时止痛作用,其机制可能与调节子宫及下丘脑AVP水平及其受体表达有关。