电针降低腹泻型肠易激综合征大鼠内脏高敏感的P2X7-ICC自噬平衡机制研究  

Study on the Mechanism of P2X7-ICC Autophagy Balance in Visceral Hypersensitivity of Diarrhea-Predominant Irritable Bowel Syndrome Rats Treated with Electroacupuncture

作　　者：翁利 熊静 邹佳珂 何昭璇 WENG Li;XIONG Jing;ZHOU Jia-ke;HE Zhao-xuan(Chengdu University of Traditional Chinese Medicine,Chengdu,Sichuan 611137,China;West China Tianfu Hospital,Sichuan University,Chengdu,Sichuan 610213,China)

机构地区：[1]成都中医药大学,四川成都611137 [2]四川大学华西天府医院,四川成都610213

出　　处：《时珍国医国药》2024年第10期2478-2485,共8页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金委员会青年基金(81804002);四川省自然科学基金青年基金(2023NSFSC1820)。

摘　　要：目的观察电针对腹泻型肠易激综合征(IBS-D)大鼠内脏高敏感及结肠P2X7、自噬相关蛋白表达水平的影响,探讨电针治疗IBS-D的可能机制。方法将24只SD大鼠随机分为空白组、模型组与电针组,每组8只。采用慢性不可预知性温和应激联合番泻叶灌胃法复制IBS-D模型。电针组交替干预单侧“足三里”“天枢”,连续14d。采用内脏痛阈值评估大鼠内脏高敏状态,Bristol评分评价大鼠腹泻情况,HE染色法观察结肠病理变化,透射电镜法观测结肠Cajal间质细胞(ICC)超微结构,免疫组织化学法检测结肠组织IL-1β、TNF-α、C-kit、Beclin1、mTOR、P2X7表达水平。结果与空白组比较,模型组大鼠内脏痛阈值显著降低(P<0.01),Bristol评分显著升高(P<0.01);结肠ICC细胞线粒体肿胀,粗面内质网轻度扩张,自噬小体减少;结肠组织IL-1β、TNF-α、C-kit、mTOR、P2X7阳性表达量显著升高(P<0.01,P<0.01,P<0.05,P<0.01,P<0.01),Beclin1阳性表达显著降低(P<0.01)。与模型组比较,电针组大鼠内脏痛阈值升高(P<0.05),Bristol评分下降(P<0.05);结肠ICC细胞结构较正常,可见明显自噬小体;结肠组织IL-1β、TNF-α、C-kit、mTOR、P2X7阳性表达显著降低(P<0.05,P<0.01,P<0.05,P<0.01,P<0.01),Beclin1阳性表达升高(P<0.05)。结论电针可以降低IBS-D大鼠的内脏高敏感性,改善IBS-D腹泻症状,其机制可能与降低结肠P2X7表达,促进结肠ICC自噬,减少ICC数量有关。Objective To observe the effects of Electroacupuncture(EA)on visceral hypersensitivity and the expression levels of colonic P2X7 and autophagy-related proteins in a rat model of diarrhea-predominant irritable bowel syndrome(IBS-D),as well as to explore the underlying mechanisms.Methods A total of 24 SD rats were randomly divided into a blank control group,a model group,and an electroacupuncture group,with 8 rats in each group,with eight rats in each group.The IBS-D model was induced by chronic unpredictable mild stress combined with intragastric administration of senna.Electroacupuncture was applied to"Zusanli"(ST36)and"Tianshu"(ST25)alternately for 14 days.The visceral hypersensitivity was evaluated by visceral pain threshold,and the severity of diarrhea was evaluated using the Bristol stool form scale.The pathological changes of colon were observed by HE staining,and the ultrastructure of interstitial cells of Cajal(ICC)was observed by transmission electron microscope.The expression levels of IL-1β,TNF-α,C-kit,Beclin1,mTOR and P2X7 in colon tissue were detected by immunohistochemistry.Results Compared with the control group,the visceral pain threshold of the model group was significantly decreased(P<0.01),and the Bristol score was significantly increased(P<0.01).The mitochondria of colonic ICC cells were swollen,the rough endoplasmic reticulum was slightly dilated,and the autophagosomes were reduced.The positive expressions of IL-1β,TNF-α,C-kit,mTOR and P2X7 in colon tissue were significantly increased(P<0.01,P<0.01,P<0.05,P<0.01,P<0.01),while the positive expression of Beclin1 was significantly decreased(P<0.01).Compared with the model group,the visceral pain threshold was increased(P<0.05),and the Bristol score was decreased(P<0.05)in the electroacupuncture group.The structure of colonic ICC cells was relatively normal,and obvious autophagosomes were observed.The positive expressions of IL-1β,TNF-α,C-kit,mTOR and P2X7 in colon tissue were significantly decreased(P<0.05,P<0.01,P<0.05,P<0.01,P<0.01),while

关 键 词：电针 腹泻型肠易激综合征 内脏高敏感 P2X7 CAJAL间质细胞 自噬 

分 类 号：R245.97[医药卫生—针灸推拿学]