隔药灸关元穴对原发性痛经大鼠疼痛反应与血清β-EP、子宫PGE2/PGF2α、脾脏NK细胞活性的影响

Effects of Herbal-cake-partitioned Moxibustion of“Guanyuan”(RN 4)on Pain Response and Serumβ-EP,Uterine PGE2/PGF2αand Spleen NK Cell Activity in Primary Dysmenoria Rats

目的观察隔药灸关元穴对原发性痛经大鼠扭体反应、血清β-内啡肽(β-EP)、子宫组织前列腺素E2(PGE2)/前列腺素F2α(PGF2α)、脾脏NK细胞活性的影响,探讨隔药灸关元治疗原发性痛经的效应机制以及与直接灸关元的效应差异。方法32只雌性Wistar大鼠随机分为空白对照组、模型组、隔药灸关元组、直接灸关元组,每组8只;皮下注射苯甲酸雌二醇联合腹腔注射缩宫素建立原发性痛经大鼠模型。隔药灸关元组采用艾炷隔药灸"关元"穴,直接灸关元组采用艾炷直接灸"关元"穴,均每日1次,每次7壮,连续10 d。末次治疗结束后,记录30 min内各组大鼠扭体次数,ELISA测定各组大鼠血清β-EP、子宫组织PGE2、PGF2α含量,MTT比色法检测脾脏NK细胞活性。结果与空白对照组比较,模型组大鼠30 min内扭体次数明显增加(P<0.01),提示造模成功;血清β-EP与子宫组织PGE2含量均显著降低(P<0.01),子宫组织PGF2α含量显著升高(P<0.01),脾脏NK细胞活性显著降低(P<0.01)。治疗结束后,与模型组比较,隔药灸关元组与直接灸关元组30 min内扭体次数明显减少(P<0.01),血清β-EP与子宫组织PGE2含量明显增加(P<0.01),子宫组织PGF2α含量明显降低(P<0.01),脾脏NK细胞活性明显升高(P<0.01)。与直接灸关元组比较,隔药灸关元组脾脏NK细胞活性更高(P<0.05)。结论隔药灸关元治疗原发性痛经作用显著,其机制可能是通过提高血清β-EP、子宫PGE2含量与脾脏NK细胞活性,降低子宫PGF2α含量实现;隔药灸关元穴与直接灸关元穴镇痛效应相当,但隔药灸对PD大鼠NK细胞活性影响优于直接灸,可能与隔药灸中药物的参与等因素有关。

Objective:To observe the effects of RN 4-HCPM on torsion response,serumβ-EP,Uterine PGE2/PGF2αand Spleen NK cell activity in Primary Dysmenoria(PD)Rats,and to explore the mechanism of therapeutic effect of RN 4-HCPM on primary dysmenorrhea,and the difference with direct moxibustion RN 4.Methods:Thirty-two Wistar female rats were equally randomized into blank control group,PD model group,RN4-HCPM group and RN 4 direct moxibustion group(n=8 rats in each).The PD model was established by subcutaneous injection of estradiol benzoate injection(0.2~0.5 mg/rat)for 10 consecutive days and intraperitoneal injection of oxytocin(2 U)24 after the last subcutaneous injection.RN 4-HCPM were used to herbal-cake(composed of Radix Angelicae Sinensis,Rhizoma Chuanxiong,Radix Paeoniae Rubra,Cortex Cinnamomi,etc.)-partitioned moxibustion,and moxa sticks were used to moxibustion“Guanyuan”point in RN 4 direct moxibustion for 7 moxa-cones every time,once daily for 10 successive days.After the end of the last treatment,the number of torsional bodies in each group was compared within 30 mins.The contents ofβ-EP,PGE2 and PGF2αin serum of each group were determined by ELISA,and the activity of spleen NK cells was detected by MTT colorimetric method.Results:Compared with the blank control group,the time of torsional bodies in the PD model group increased significantly within 30 min(P<0.01),indicating that the model was successful;serumβ-EP and uterine tissue PGE2 levels were significantly reduced(P<0.01),the content of uterine tissue PGF2αwas significantly increased(P<0.01),and the activity of NK cells in the spleen was significantly decreased(P<0.01).After treatment,compared with the model group,the number of torsional bodies within 30 min in the group with RN 4-HCPM and RN 4 direct moxibustion group reduced clearly(P<0.01),the content ofβ-EP and PGE2 in serum was significantly increased(P<0.01),the content of PGF2αin uterine tissues was significantly decreased(P<0.01),and the activity of NK cells in the spleen was significantly increased(P<0.01).Compared with RN4 direct moxibustion group,NK cell activity of spleen was more significant in the RN 4-HCPM group(P<0.05).Conclusion:RN 4-HCPM treats primary dysmenorrhea with obvious therapeutic effect,the treatment mechanism may be through increasing serumβ-EP,uterus tissue content of PGE2 and spleen NK cell activity,reducing the content of uterus tissue PGF2α.The analgesic effect of RN 4-HCPM is similar to that of direct moxibustion RN 4,which may be related to the involvement of the drugs in the medicated moxibustion and different moxibustion.