基于EP2/NLRP3信号通路探讨雀啄灸对原发性痛经大鼠的影响

Effect of sparrow-pecking moxibustion on primary dysmenorrhea rats based on EP2/NLRP3 signal pathway

目的探讨雀啄灸对原发性痛经大鼠前列腺素E受体2(EP2)/Nod样受体蛋白3(NLRP3)的调控作用及相关抗炎机制。方法24只雌性SD大鼠按照随机数字表法分为空白组、模型组、雀啄灸组、激动剂组,每组6只。除空白组外其余三组建立原发性痛经模型;空白组每日注射同等剂量生理盐水。空白组及模型组不予治疗;雀啄灸组每日造模后,选取三阴交和关元穴进行治疗;激动剂组每日造模后腹腔注射Treprostinil,之后进行雀啄灸治疗,治疗方法同雀啄灸组。观察各组扭体潜伏时间、扭体次数;HE染色观察子宫病理情况;免疫组织化学法检测EP2、NLRP3、caspase-1蛋白表达。结果与空白组比较,模型组大鼠潜伏时间缩短、扭体次数增多(P<0.01);与模型组比较,雀啄灸组潜伏时间延长、扭体次数减少(P<0.01);与雀啄灸组比较,激动剂组潜伏期缩短、扭体次数增多(P<0.01)。空白组子宫组织形态规则,肌细胞间质内少许淋巴细胞,无中性粒细胞浸润;模型组、激动剂组子宫小血管有扩张及充血表现,淋巴细胞与嗜酸性粒细胞等炎症细胞呈灶团样向外浸润明显;雀啄灸组间质内充血水肿及炎症细胞浸润程度较模型组减轻,肌壁间可见少许炎症细胞浸润,仅见少许小血管充血。与空白组比较,模型组EP2、NLRP3、caspase-1蛋白表达升高(P<0.01);与模型组比较,雀啄灸组EP2、NLRP3、caspase-1蛋白表达降低(P<0.01);与雀啄灸组比较,激动剂组NLRP3、caspase-1蛋白表达升高(P<0.01),雀啄灸组与激动剂组EP2蛋白表达比较,差异无统计学意义(P>0.05)。结论雀啄灸可以改善子宫炎症细胞浸润的情况,通过调控EP2/NLRP3信号通路调节痛经大鼠炎症反应。

Objective To investigate the regulation of prostaglandin E receptor 2(EP2)/Nod-like receptor protein 3(NLRP3)and its anti-inflammatory mechanism in primary dysmenorrhea rats by sparrow-pecking moxibustion.Methods A total of 24 female SD rats were divided into blank group,model group,sprrow-pecking moxibustion group,and agonist group,with six rats in each group.Except blank group,the primary dysmenorrhea model was established for the other three groups,blank group received the same dose of normal saline daily.Blank group and model group were not treated;in sparrow-pecking moxibustion group,sānyīnjiāo and guanyuán acupoints were selected for treatment after daily modeling;agonist group was intraperitoneally injected with Treprostinil daily after modeling,followed by sparrow-pecking moxibustion treatment,which was the same as the sparrow-pecking moxibustion group.The latent time and number of torsion were observed in each group;HE staining was used to observe the pathological conditions of uterus;the expressions of EP2,NLRP3,and caspase-1 proteins were detected by immunohistochemistry.Results Compared with blank group,the latent time was shortened and the number of torsion was increased in model group(P<0.01);compared with model group,the latent time was prolonged and the number of torsion was decreased in sparrow-pecking moxibustion group(P<0.01);compared with sparrow-pecking moxibustion group,the latent time was shortened and the number of torsion was increased in agonist group(P<0.01).In blank group,the uterine tissue morphology was regular,there were a few lymphocytes in the interstitium of muscle cells,and no neutrophil infiltration;in model group and agonist group,small uterine vessels showed dilation and hyperemia,and inflammatory cells such as lymphocytes and eosinophils were infiltrated outwards as focal groups;in sparrow-pecking moxibustion group,the degree of interstitial hyperemia and edema and infiltration of inflammatory cells were less than that in model group,a few inflammatory cells infiltrated between muscle walls and only a few small blood vessels hyperemia were observed.Compared with blank group,the protein expressions of EP2,NLRP3,and caspase-1 were increased in model group(P<0.01);compared with model group,the protein expressions of EP2,NLRP3,and caspase-1 were decreased in sparrow-pecking group(P<0.01);compared with sparrow-pecking group,the protein expressions of NLRP3 and caspase-1 were increased in agonist group,while the protein expression of EP2 in sparrow-pecking group and agonist group was not significantly different(P>0.05).Conclusion Sparrow-pecking moxibustion can improve the infiltration of inflammatory cells in the uterus,and regulate the inflammatory response of dysmenorrhea rats by regulating EP2/NLRP3 signal pathway.