电针对IBS内脏痛大鼠脊髓GFAP、P2X3受体的调节作用

Regulation of GFAP and P2X3 Receptors in the Spinal Cord of IBS Rats with Visceral Pain

目的探讨脊髓星形胶质细胞和P2X3受体在电针缓解肠易激综合征(Irritable Bowel Syndrome,IBS)大鼠内脏痛的调节作用机制。方法采用直结肠球囊扩张(Colorectal dilatation,CRD)刺激诱导IBS慢性内脏痛模型。将清洁级SD(Sprague-Dawley)雄性新生大鼠随机分为正常组和造模组。通过行为学评分和结肠形态学方法鉴定模型制备成功后将造模组随机分为模型组、电针组、氟代柠檬酸组。电针组取双侧上巨虚穴(S37)、天枢穴(S25)进行电针治疗,针刺深度为5 mm,30 min/次,每日1次,连续治疗7天。氟代柠檬酸组鞘内注射氟代柠檬酸10μL,每周3次。治疗结束后,采用腹部撤回反射(Abdominal Withdrawal Reflex,AWR)评分评估电针对IBS内脏痛大鼠行为学影响;采用HE染色观察IBS内脏痛大鼠的结肠组织病理形态学变化;采用免疫组织化学、Western Blot等方法检测脊髓中星形胶质细胞标志物胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)和P2X3受体蛋白表达。结果与正常组相比,模型组大鼠AWR评分显著升高,脊髓中GFAP和P2X3受体表达水平明显增多;与模型组比较,电针组、氟代柠檬酸组大鼠AWR评分明显降低,脊髓中GFAP、P2X3受体的表达明显降低。结论电针能够有效缓解IBS大鼠内脏痛敏,其镇痛效应可能与其抑制脊髓GFAP和P2X3受体表达有关。

Objective To explore the regulatory mechanism of spinal astrocytes and P2 X3 receptors in the alleviation of visceral pain in IBS rats with electroacupuncture.Methods The colorectal dilatation(CRD)stimulation was used to induce the chronic visceral pain model of IBS.The clean-grade SD(Sprague-Dawley)male newborn rats were randomly divided into normal group and model group.After the model was successfully prepared through behavioral scoring and colon morphology,the model was randomly divided into model group,electroacupuncture group,and fluorocitric acid group.In the electroacupuncture group,the bilateral Shangjuxu(S37)and Tianshu acupoints(S25)were treated with electro-acupuncture.The acupuncture depth was 5 mm,30 min/time,once a day,for 7 consecutive days.In the fluorocitric acid group,10 ul of fluorocitric acid was injected intrathecally,3 times a week.After treatment,the AWR score was used to assess the behavioral effects of electroacupuncture in rats with IBS visceral pain;HE staining was used to observe the pathological changes of colon tissue in rats with IBS visceral pain;immunohistochemistry,Western Blot and other methods were used to detect GFAP and GFAP in the spinal cord.P2 X3 receptor protein expression.Results Compared with the normal group,the AWR scores of rats in the model group were significantly higher,and the expression levels of GFAP and P2 X3 receptors in the spinal cord were significantly increased;compared with the model group,the AWR scores of rats in the electroacupuncture and fluorocitric acid groups were significantly lower,and the expression of GFAP and P2 X3 receptors in the spinal cord were significantly lower.Conclusion Electroacupuncture can effectively alleviate visceral hyperalgesia in IBS rats,and its analgesic effect may be related to its inhibition of spinal cord GFAP and P2 X3 receptor expression.