大建中汤对肠易激综合征内脏痛大鼠中枢敏化的作用机制  

Mechanism of Dajianzhong Decoction in regulating central sensitization in rats with irritable bowel syndrome visceralgia

作　　者：武静 王慧 田维毅 蔡琨 李尧锋 杨莎莎 WU Jing;WANG Hui;TIAN Weiyi;CAI Kun;LI Yaofeng;YANG Shasha(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;The First AffiliatedHospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州中医药大学基础医学院,贵阳550025 [2]贵州中医药大学第一附属医院,贵阳550001

出　　处：《中华中医药杂志》2025年第1期375-380,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82260855,No.81960821);贵州省基础研究计划(自然科学)项目(No.黔科合基础-ZK[2022]一般495);贵州省卫生健康委科学技术基金项目(No.gzwkj2023-503)。

摘　　要：目的:研究肠易激综合征(IBS)内脏痛大鼠前扣带回皮质(ACC)中枢敏化物质、兴奋性氨基酸、细胞因子、能量代谢物质的表达及大建中汤的干预作用。方法:通过腹腔注射卵清白蛋白、母婴分离、乙酸灌肠等方式制备IBS内脏痛大鼠模型,将模型大鼠随机分为模型组(灌服等量0.9%氯化钠溶液,每天1次,持续2周)和大建中汤组(灌胃大建中汤水浸液10.8 g/kg,每天1次,持续2周),另设正常组(灌服等量0.9%氯化钠溶液,每天1次,持续2周),每组10只。采用腹部撤离反射(AWR)评估各组大鼠内脏疼痛敏感度;采用免疫印迹试验法检测大鼠脑ACC内中枢敏化物质酪氨酸蛋白激酶类固醇受体辅激活因子(Src)、N-甲基-D-天冬氨酸受体2B亚单位(NMDAR-2B)、血清缝隙连接蛋白-1(Panx1)的蛋白表达;ELISA法测定大鼠ACC组织内P物质(SP)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的含量;微透析分析仪检测大鼠脑ACC组织透析液中能量代谢物质如乳酸(Lac)、丙酮(Pyr)、葡萄糖(Gluc)的表达;高效液相色谱法(HPLC)检测大鼠ACC组织透析液中兴奋性氨基酸神经递质D-丝氨酸(D-Ser)、甘氨酸(Gly)、谷氨酸(Glu)的含量。结果:与正常组比较,模型组AWR评分于60、40、20 mmHg(1 mmHg≈0.133 kPa)压力下显著增加(P<0.01),ACC组织中Panx1、Src、NMDAR-2B蛋白表达显著上升(P<0.01),TNF-α、IL-1β、SP含量显著升高(P<0.01),透析液中Glu、D-Ser、Gly、Gluc、Lac含量显著升高(P<0.01)。与模型组比较,大建中汤组AWR评分于60、40、20mmHg压力下显著下降(P<0.01),ACC组织中Panx1、Src、NMDAR-2B蛋白表达显著下降(P<0.01),TNF-α、IL-1β、SP含量显著降低(P<0.01),透析液中Glu、D-Ser、Gly、Gluc、Lac含量降低(P<0.01)。结论:大建中汤调节IBS内脏痛中枢敏化的机制可能与其抑制大鼠ACC细胞外液中Glu、D-Ser、Gly、Gluc、Lac的含量,降低Panx1、Src和NMDAR-2B蛋白表达有关。Objective:To study the expressions of central sensitization substances,excitatory amino acid(EAA),cytokines and energy metabolites in anterior cingulate cortex(ACC) of rats with visceralgia of irritable bowel syndrome(IBS) and the intervention effect of Dajianzhong Decoction(DD).Methods:Prepare model rats with IBS visceralgia through intraperitoneal injection of ovalbumin,maternal separation,and acetic acid enema.And then,separate them randomly into model group and DD group.Each rat from the model group was drenched with the same amount of normal saline,once a day for 2 weeks,and each rat from the DD group was drenched with Dajianzhong Decoction 10.8 g/kg,once a day for 2 weeks.Another control group was made up of unaffected rats,each of which was drenched with the same amount of normal saline,once a day for 2 weeks.The number of rats in each of the 3 groups was 10.After that,evaluate and study the visceralgia sensitivity of each rat from every group through abdominal withdrawal reflex(AWR).Western Blot was applied to detect the expressions of central sensitization substances in rat ACC,such as tyrosine protein kinase Src,N-methyl-D-aspartate receptor subunit 2B(NMDAR-2B) and serum pannexin-1(Panx1).The concentrations of substance P(SP),interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in rat ACC tissues were measured through ELISA.A microdialysis analyzer was used to detect the expressions of energy metabolites in the dialysate of rat ACC tissues,such as lactic acid(Lac),pyruvic acid(Pyr),and glucose(Gluc).High performance liquid chromatography(HPLC)was used to determine the concentrations of EAA neurotransmitters in the dialysate of rat ACC tissues,including D-serine(D-Ser),glycine(Gly) and glutamate(Glu).Results:Compared with control group,the model group AWR scores were higher under the pressure of 60,40,and 20 mmHg(1 mmHg≈0.133 kPa)(P<0.01);protein expressions of Panx1,Src and NMDAR-2B in ACC tissues were higher(P<0.01);concentrations of TNF-α,IL-1β and SP were higher(P<0.01);concentrations of

关 键 词：肠易激综合征 内脏痛 氨基酸类神经递质 乳酸 中枢敏化 大建中汤 

分 类 号：R285.5[医药卫生—中药学]