芪地通便方对慢传输型便秘小鼠钙离子/结肠钙调蛋白/肌球蛋白轻链激酶信号通路的影响  

Effect of Qidi Tongbian Decoction on Ca 2+/CaM/MLCK pathway in mice with slow transit constipation

作　　者：刘冠茜 李军祥[1] 毛堂友 石磊[1] 王佳丽 蒋婉婷 庞家乐 杨思邈 陈润花[1] LIU Guanxi;LI Junxiang;MAO Tangyou;SHI Lei;WANG Jiali;JIANG Wanting;PANG Jiale;YANG Simiao;CHEN Runhua(Gastroenterology Department,Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学东方医院脾胃肝胆科,100078

出　　处：《环球中医药》2025年第4期701-710,共10页Global Traditional Chinese Medicine

基　　金：国家中医药管理局高水平中医药重点学科中西医结合临床(消化病学)(zyyzdxk-2023271);北京市中医药科技发展资金项目(JJ-2020-37);北京中医药大学2023年度基本科研业务费项目“揭榜挂帅项目”(2023-JYB-JBQN-015)。

摘　　要：目的基于网络药理学及动物实验探究芪地通便方对慢传输型便秘(slow transit constipation,STC)的作用机制。方法通过数据库检索获得芪地通便方药物活性成分、作用靶点、疾病靶点,进行京都基因与基因组百科全书通路富集分析,预测芪地通便方治疗STC的作用通路。通过盐酸洛哌丁胺灌胃以建立小鼠STC模型进行实验验证。共30只SPF级雄性BALB/C小鼠,随机分为空白组、模型组、阳性药组和(芪地通便方)低、中、高剂量组,每组5只,并进行相应药物干预。检测计算小鼠粪便含水率、小肠推进率,观察记录结肠组织病理学变化,进行分子生物学检测并计算结肠钙离子(calciumion,Ca^(2+))浓度,结肠钙调蛋白(calmodulin,CaM)、肌球蛋白轻链激酶(myosin light-chain kinase,MLCK)mRNA及蛋白相对表达量、磷酸化肌球蛋白轻链(phospho-myosin light chain,p-MLC)蛋白相对表达量。结果网络药理学筛选出芪地通便方活性成分74个,疾病与药物交集靶点510个,最终获得钙离子信号通路等芪地通便方在STC治疗中具有较高预测概率的信号通路。动物实验方面,与空白组相比,模型组小鼠的粪便含水率、小肠推进率、Ca^(2+)浓度、CaM、MLCK的mRNA及蛋白表达、p-MLC蛋白表达均明显下降(P<0.05);结肠组织出现黏膜充血肿胀,固有层内腺体萎缩、紊乱,杯状细胞数量显著减少等病理变化。与模型组比较,阳性药组、芪地通便方各剂量组小鼠的粪便含水率、小肠推进率、Ca^(2+)浓度、CaM、MLCK的mRNA及蛋白表达和p-MLC蛋白表达均明显升高(P<0.05);结肠组织黏膜层结构完整,腺体排列密集,杯状细胞数量明显增多。结论芪地通便方具有改善STC症状的药效学作用,其作用机制可能与激活Ca^(2+)/CaM/MLCK信号通路的活性相关。Objective To investigate the mechanism of Qidi Tongbian Decoction in the treatment of slow transit constipation(STC)based on network pharmacology and animal experiments.Methods The pharmacological active ingredients and corresponding targets of Qidi Tongbian Decoction,as well as the disease targets associated with STC,were retrieved from public databases.KEGG pathway enrichment analysis was conducted to predict the therapeutic pathways through which Qidi Tongbian Decoction exerts its effects on STC.30 SPF-grade male BALB/C mice were randomly divided into untreated control group,model group,Qidi Tongbian Decoction low-dose group,Qidi Tongbian Decoction medium-dose group,Qidi Tongbian Decoction high-dose group,and positive control group(n=5 per group),and subjected to corresponding drug interventions.During the experiment,stool water content and intestinal transit rate were measured and calculated.Histopathological changes in the colon were observed and recorded.Molecular biological assessments were conducted to measure colonic calcium ion(Ca^(2+))concentration,as well as mRNA and protein expression levels of calmodulin(CaM),myosin light chain kinase(MLCK),and phosphorylated myosin light chain(p-MLC).Results From the network pharmacology analysis,74 active ingredients of Qidi Tongbian Decoction and 510 disease-drug intersection targets were identified.Ultimately,calcium pathway was selected for further experimental research as it had a high predictive probability in STC treatment.In animal experiments,compared with the untreated control group,the model group exhibited was significant decreased in stool water content,intestinal transit rate,Ca^(2+)concentration,mRNA and protein expression levels of CaM,MLCK,and p-MLC(P<0.05).Pathological changes in the colon tissue included mucosal congestion and swelling,atrophy and disorder of glands in the lamina propria,and a significant reduction in the number of goblet cells.Compared with the model group,the positive control group and all dose of Qidi Tongbian Decoction groups

关 键 词：慢传输型便秘 芪地通便方 钙离子/结肠钙调蛋白/肌球蛋白轻链激酶信号通路 网络药理学 实验验证 

分 类 号：R285.5[医药卫生—中药学]