温和灸对溃疡性结肠炎大鼠天枢穴P2RX_7的作用机理研究

Effect of Warming Moxibustion on P2RX_7 Regulation in Tianshu Acupoint Skin of Rats with Ulcerative Colitis

目的:溃疡性结肠炎(UC)是艾灸治疗的有效病症,本研究从艾灸作用的始动环节与穴位响应密切相关的角度,探讨温和灸的对穴区P2X嘌呤受体7表达的影响。方法:将20只SD雄性大鼠随机分成正常组6只、模型组14只。采用自由饮用4%葡聚糖硫酸钠(DSS)法建立UC模型,连续饮用7天后,取2只大鼠进行模型鉴定,造模成功后将剩下的12只大鼠随机分成UC组、UC+温和灸组,第8天改为自由饮用1%DSS溶液,第8天开始采用温和灸天枢穴(双)干预7天,每天10 min。疗程结束后,称量大鼠体质量、取双侧天枢穴皮肤,采用HE染色观察病理组织学形态变化,采用免疫组化法对P2RX7的蛋白检测,从分子水平观察温和灸对天枢穴皮肤的作用;采用q RT-PCR的方法检测P2RX7在天枢穴皮肤中的表达。结果:与正常组比较,UC组体质量于第4天开始明显降低(P<0.05),天枢穴P2RX7基因的m RNA表达水平降低(P<0.05),天枢穴皮肤中主要分布在真皮层皮脂腺中的P2RX7明显减少(P<0.05)。与UC组比较,UC+温和灸组在第8天明显增加(P<0.05),天枢穴P2RX7基因m RNA表达水平增高(P<0.05),天枢穴皮肤P2RX7主要分布在皮脂腺中的明显增加(P<0.05)。温和灸干预后所增加体重与天枢穴P2RX7免疫组化IOD值的相关系数r=0.823(P<0.05)。结论:温和灸能明显增加UC大鼠体重,其起效的机制可能与上调天枢穴位皮肤P2X嘌呤受体7密切相关。

Moxibustion is an effective treatment for ulcerative colitis（UC）. Structure and function of acupoints are closely related to the response to moxibustion for patients. This study investigated purinergic receptor P2X7 m RNA and protein expressions of Tianshu acupoint under warming moxibustion in UC rats. The 20 male SD rats were randomly divided into two groups： the normal group（n = 6） and the model group（n = 14）. The model group rats were fed with 4% dextran sulfate sodium（DSS） for 7 days. Among the 14 rats, 2 rats were taken for identification of successful UC model before moxibustion treatment. The rest of 12 rats in the model group were randomly divided into the UC group, the UC and warming moxibustion group, and fed with 1% DSS solution. On the 8th day, warming moxibustion at bilateral Tianshu had been used in the UC and warming moxibustion group for 7 days, 10 minutes per day. Rats were weighed every day. After 7 days warming moxibustion, the Tianshu acupoint skins of both sides were taken for HE staining and immunohistochemistry（IHC）. The expression of P2X7 on Tianshu acupoint skin was detected by q RT-PCR. The results showed that compared with the normal group, the rat body weight decreased from the 4th day in the UC group（P〈 0.05）; and gene P2RX7 expression level also significantly decreased in the Tianshu acupoint skin（P〈 0.05）; P2RX7 in the sebaceous glands of the dermis significantly decreased on Tianshu acupoint skin（P〈 0.05）. Compared with the model group, the rat body weight increased from 8th day in the UC and warming moxibustion group（P〈 0.05）; and gene P2RX7 expression level also significantly increased in the Tianshu acupoint skin（P〈 0.05）; P2RX7 in the sebaceous glands of the dermis significantly increased on Tianshu acupoint skin（P〈 0.05）. It was positive correlation, r = 0.823（P〈 0.05）, between the body weight increased by warming moxibustion and the integrated option density（IOD） value of P2RX7 in Tianshu acupoint skin in UC. It was concluded that warming moxibustion could significantly increase the body weight of UC rats. The underlying mechanism may be closely related to the upregulation of purinergic receptor P2X7 expression in Tianshu acupoint skin.