电针对慢性肛门直肠痛模型大鼠痛觉敏化及脊髓胶质细胞的影响  

Effects of electroacupuncture on pain sensitization and spinal cord glial cells in rats with chronic anorectal pain

作　　者：曾禹沙 李婧婧 许新意 薛雅红[2] ZENG Yusha;LI Jingjing;XU Xinyi;XUE Yahong(Graduate School of Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学研究生院,江苏南京210023 [2]南京中医药大学附属南京中医院,江苏南京210001

出　　处：《陕西中医》2023年第10期1331-1336,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金资助项目(82004362)。

摘　　要：目的:观察慢性肛门直肠痛(CAP)大鼠S 2-3脊髓背角中胶质细胞活化情况及电针对其干预作用,探讨脊髓胶质细胞活化是否引起CAP及参与电针镇痛。方法:18只SD大鼠随机分为正常组、模型组和电针组。正常组直肠注入0.9%氯化钠溶液,模型组及电针组注入辣椒素(浓度:10-4 mmol/L),每间隔5 min注入1次,共注入4次;电针组造模完成且大鼠清醒2 h后予以针刺八髎穴干预。观察大鼠内脏疼痛相关行为次数及机械痛阈值变化;采用苏木素-伊红(HE)染色观察直肠组织病变;免疫荧光法检测脊髓小胶质细胞标志物OX42和星形胶质细胞标志物GFAP表达强度、酶联免疫吸附试验(ELISA)检测脊髓和血清中IL-1β、TNF-α和IL-6的含量。结果:与正常组相比,模型组大鼠内脏痛行为明显增加,大鼠尾部、后足底及腹部机械痛阈值上升(P<0.01);与模型组相比,电针组内脏痛行为减少,大鼠尾部、后足底、腹部机械痛阈值降低(P<0.01)。HE染色示正常组直肠黏膜下组织无水肿现象,模型组直肠黏膜下组织出现明显水肿病变,电针干预后,大鼠直肠黏膜下组织水肿相比模型组明显减轻;与正常组相比,模型组大鼠脊髓中小胶质细胞标志物OX42和星形胶质细胞标志物GFAP的阳性表达显著增加(P<0.01);与模型组相比,电针组大鼠脊髓中标志物OX42和GFAP的阳性表达降低,差异具有统计学意义(P<0.05)。与正常组相比,模型组大鼠脊髓和血清中IL-1β、TNF-α和IL-6水平上升(P<0.01);与模型组相比,电针组大鼠脊髓及血清中IL-1β、TNF-α和IL-6水平均下降,差异具有统计意义(P<0.05)。结论:脊髓胶质细胞活化及促炎因子IL-1β、TNF-α和IL-6释放与CAP发生密切相关,电针治疗可通过抑制脊髓胶质细胞活化及降低炎症因子IL-1β、TNF-α和IL-6水平有效缓解慢性肛门直肠疼痛。Objective:To observe the activation of glial cells in S 2-3 spinal dorsal horn of rats with chronic anorectal pain(CAP)and the intervention effect of electroacupuncture,and to explore whether the activation of spinal glial cells causes CAP and participates in electroacupuncture analgesia.Methods:A total of 18 SD rats were randomly divided into the normal group,the model group and the electroacupuncture group.The normal group was injected with 0.9%sodium chloride solution into the rectum,and the model group and the electroacupuncture group were injected with capsaicin(concentration:10-4 mmol/L),once every five minutes,four times in total;the rats in the electroacupuncture group were intervened by acupuncture at Baliao point 2 hours after the modeling was completed and the rats were awake.The changes of visceral pain-related behaviors and mechanical pain threshold in rats were observed.Hematoxylin-eosin(HE)staining was used to observe rectal tissue lesions.The expression of microglia marker OX42 and astrocyte marker GFAP in spinal cord was detected by immunofluorescence.The levels of IL-1β,TNF-αand IL-6 in spinal cord and serum were detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the normal group,the visceral pain behavior of the rats in the model group was significantly increased,and the mechanical pain threshold of the tail,hindfoot and abdomen of the rats was increased(P<0.01).Compared with the model group,the visceral pain behavior of the electroacupuncture group was reduced,and the mechanical pain threshold of the tail,hindfoot and abdomen of the rats was reduced(P<0.01).HE showed that there was no edema in the rectal submucosal tissue of the normal group,and there was obvious edema in the rectal submucosal tissue of the model group.After electroacupuncture intervention,the edema of the rectal submucosal tissue of the rats was significantly reduced compared with the model group.Compared with the normal group,the positive expression of microglia marker OX42 and astrocyte marker GFAP

关 键 词：慢性肛门直肠痛 电针 脊髓胶质细胞 白细胞介素-1Β 肿瘤坏死因子-α 白细胞介素-6 

分 类 号：R574.8[医药卫生—消化系统]