隔药灸“命门”穴对痛经模型大鼠血清雌激素、孕激素水平及其受体表达的影响

Effect of Drug-paste Separated Moxibustion on Expression of Estrogen,Progestogen and Their Endometrial Receptor mRNA in Rats with Primary Dysmenorrhea

目的:通过观察隔药灸"命门"穴对痛经模型大鼠血清雌激素(E2)和孕激素(P)水平及其受体表达的影响,探讨隔药灸"命门"穴治疗原发性痛经的作用机制。方法:雌性SD大鼠随机分为空白组、模型组、药物组、针刺组、隔药灸组,每组20只。以缩宫素建立大鼠原发性痛经模型。药物组用加味益母草膏按每日80g/kg灌胃给药;针刺组针刺"命门"穴,每次留针30min;隔药灸组用理中汤加减制成的药饼置于"命门"穴上,其上置小艾炷点燃施灸,连灸6壮。各组均于股部皮下注射苯甲酸雌二醇的第5天开始治疗,每日1次,连续7d。比较各组大鼠的扭体反应潜伏期和30min内扭体数,并用ELISA法检测各组大鼠血清E2和P的水平,用RT-PCR检测各组大鼠子宫内膜雌激素受体(ER)及孕激素受体(PR)mRNA水平。结果:与空白组比较,模型组扭体次数较多,首次扭体潜伏期缩短(P<0.01);与模型组相比,药物组、针刺组以及隔药灸组均能明显延长首次扭体潜伏期,且能减少30min内扭体反应次数(均P<0.01);与药物组相比,针刺组及隔药灸组均能明显减少30min内扭体反应次数(P<0.01);隔药灸组30min内扭体次数最少(P<0.01)。与空白组比较,模型组血清E2含量显著增加,P含量显著降低(P<0.01);与模型组比较,药物组、针刺组以及隔药灸组E2显著降低,P显著增加(P<0.01);与药物组比较,针刺组与隔药灸组E2显著降低,P显著增加(P<0.01);与针刺组比较,隔药灸组E2显著降低,P显著增加(P<0.01)。与空白组比较,模型组大鼠子宫内膜ER mRNA表达显著增加(P<0.01),PR mRNA表达降低(P<0.05);与模型组比较,药物组大鼠子宫内膜ER mRNA表达显著降低,PR mRNA表达增加(P<0.05,P<0.01);与药物组比较,针刺组与隔药灸组ER mRNA表达降低,PR mRNA表达增加(P<0.05);与针刺组比较,隔药灸组ER mRNA表达降低,PR mRNA表达增加(P<0.05)。结论:隔药灸治疗原发性痛经可能与降低痛经大鼠血清E2、子宫内膜ER mRNA表达,增加血清P、子宫内膜PR mRNA表达有关。

Objective To observe the effect of drug-paste separated moxibustion of ＂Mingmen＂ （GV 4） on the levels of serum estrogen （E2） and progesterone （P） and their endometrial receptor mRNA expression in rats with primary dysmenorrhea in order to investigate its mechanism underlying improvement of primary dysmenorrhea. Methods A total of 100 female SD rats were randomized into control, model, medication, acupuncture and moxibustion groups, with 20 rats in each group. Primary dys- menorrhea model was established by subcutaneous injection of Benzestrofol for 10 days and intraperitoneal injection of Oxytocin for 1 d. Rats of the medication group were fed with extractum leonuri inspissatum （8 g/100 g） and those of the moxibustion group treated with drug-paste separated moxibustion at ＂Mingmen＂ （GV 4）. For rats of the acupuncture group, a filiform needle was in- serted into GV 4, manipulated for a while and retained for 30 rain. The treatment of the latter 3 groups was conducted once daily for 7 days. The rat＇s body-writhing latency and times during 30 min were recorded. The contents of serum E2 and P were detectedby ELISA, and the expression of estrogen receptor （ER） mRNA and progesterone receptor （PR） mRNA in the endometrium was determined by quantitative real-time （RT）-PCR. Results （The body-writhing latency was shorter and the writhing times were more in the model group than in the control group（ P〈0.01 ）. Compared with the model group, the body-writhing latency was sig-nificantly increased and the writhing times were obviously decreased in the medication, acupuncture and moxibustion groups （ P〈 0.0 1 ）. There were no significant differences among the medication, acupuncture and moxibustion groups in the body-writhing latency （P〈0.05）o but the body-writhing numbers of the acupuncture and moxibustion groups were markedly lower than that of the medication group（ P〈0.01 ）. （2）Compared with the control group, serum E2 content and endometrial ER mRNA expression le vel were significantly increased, and serum P content and endometrial PR mRNA level evidently decreased in the model group （P〈0.0 , P〈0.05）. In comparison with the model group, serum E2 contents and endometrial ER mRNA expression levels were considerably down-regulated, and serum P contents and endometrial PR mRNA expression levels markedly up-regulated in the medication, acupuncture and moxibustion groups （ P〈0.01, P〈0.05）. The effects of the moxibustion group were significantly su-perior to those of the acupuncture and medication groups, and those of the acupuncture group were also significantly superior to those of the medication group in lowering E and endometrial ER mRNA levels, and raising serum P and endometrial PR mRNA ex pression levels （ P〈0.01, P〈0.05）. Conclusion Drug-paste separated moxibustion of GV 4 is effective in relieving pain in primary dysmenorrheal rats, which is probably associated with its effects in down-regulating serum E2 content and endometrial ER mRNA expression, and up-regulating serum P and endometrial PR mRNA expression levels.