基于TRPV4介导BDNF/TrkB/CREB信号通路探究畅舟通便方对功能性便秘合并抑郁样行为大鼠的干预机制  

Exploring Intervention Mechanism of Changzhou Tongbian Formula on Functional Constipation Combined with Depressive-like Behaviors in Rats Based on TRPV4-mediated BDNF/TrkB/CREB Signaling Pathway

作　　者：姚玲 权隆芳[1] 谢振年[1] 戴洁 刘秋宁 李永海[2] 贾小强[1] YAO Ling;QUAN Longfang;XIE Zhennian;DAI Jie;LIU Qiuning;LI Yonghai;JIA Xiaoqiang(Department of Anorectal,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Department of Anorectal,Hefei First People’s Hospital,Hefei 230001 Anhui,China)

机构地区：[1]中国中医科学院西苑医院肛肠科,北京100091 [2]合肥市第一人民医院肛肠科,安徽合肥230001

出　　处：《中药新药与临床药理》2024年第9期1289-1298,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：安徽省重点研究与开发计划项目(202104j07020048);中国中医科学院科技创新工程项目(CI2021A01910,CI2021A00911);西苑医院院级课题(xyzy0201-07,XYZX0203-03)。

摘　　要：目的探究畅舟通便方对功能性便秘(functional constipation,FC)合并抑郁大鼠的干预机制。方法采用盐酸小檗碱法联合慢性不可预知温和刺激建立功能性便秘合并抑郁样行为大鼠模型,随机分为空白组、模型组、乳果糖组及畅舟通便方高、中、低剂量组,干预5周。分别于干预前、干预后计算粪便含水量;干预结束后计算小肠推进率;分别于干预前、干预后采用强迫游泳实验、糖水偏好实验评估大鼠的抑郁样行为水平;HE染色观察结肠黏膜形态;尼氏染色观察海马CA1、DG区锥体细胞形态和数量;Western Blot法检测各组大鼠结肠组织的TRP离子通道香草香素4型(TRPV4)及海马脑源性神经营养因子(BDNF)、酪氨酸激酶受体(TrkB)、丝裂原活化蛋白激酶(MAPKs)、环磷腺苷效应元件结合蛋白(CREB)蛋白相对表达量。结果与空白组比较,模型组大鼠的粪便含水率明显减少(P<0.001),小肠推进率明显降低(P<0.01),糖水偏好指数明显下降(P<0.001),海马CA1区、DG区的锥体细胞数量明显减少(P<0.001),结肠TRPV4以及海马BDNF、TrkB、MAPK、CREB蛋白含量均明显下降(均P<0.001)。与模型组比较,畅舟通便方高、中、低剂量组及乳果糖组大鼠的粪便含水率均明显增高(P<0.001,P<0.01);畅舟通便方高、低剂量组及乳果糖组的小肠推进率明显增高(P<0.01,P<0.05);畅舟通便方高、中、低剂量组及乳果糖组的糖水偏好指数明显升高(P<0.01,P<0.05);结肠病理无异常;畅舟通便方高、中、低剂量组及乳果糖组的海马CA1、DG区锥体细胞数目明显增多(P<0.01,P<0.001);畅舟通便方高、中、低剂量组及乳果糖组的结肠TRPV4含量明显增加(P<0.001),海马BDNF、TrkB、MAPK、CREB蛋白含量均明显增加(P<0.05,P<0.01,P<0.001)。结论畅舟通便方干预后可明显提高功能性便秘合并抑郁模型大鼠结肠的TRPV4含量,增加粪便含水量,促进肠蠕动,可改善便秘;同时TRPV4可激活BDNF/TrkB/MAPK�Objective To explore the intervention mechanism of Changzhou Tongbian Formula on functional constipation(FC)combined with depression in rats.Methods A rat model of FC combined with depressive-like behaviors was established by using gastric gavage of berberine hydrochloride and chronic unpredictable mild stress.The model rats were randomly divided into blank group,model group,lactulose group,and Changzhou Tongbian Formula high-,medium-,and low-dose groups,and the intervention was carried out for five weeks.The fecal water content was calculated before and after intervention.The small intestinal propulsion rate was calculated after intervention.The depressive-like behavior levels of rats were evaluated by forced swimming test and sucrose preference test before and after intervention.The colonic mucosal morphology was observed by HE staining.The morphology and number of pyramidal cells in the hippocampal CA1 and DG regions were observed by Nissl staining.The relative protein expression levels of the transient receptor potential vanilloid 4(TRPV4)in colonic tissue,as well as brain-derived neurotrophic factor(BDNF),tyrosine kinase receptor B(TrkB),mitogen-activated protein kinase(MAPK),and cAMP-response element binding protein(CREB)in the hippocampus of rats in each group were detected by Western Blot method.Results Compared with the blank group,the fecal water content of rats in the model group was significantly reduced(P<0.001),the small intestinal propulsion rate was significantly lower(P<0.01),the sucrose preference index was significantly decreased(P<0.001),the number of pyramidal cells in the CA1 and DG regions of the hippocampus was significantly reduced(P<0.001),and the TRPV4 in the colon and the BDNF,TrkB,MAPK,and CREB protein contents in the hippocampus were all significantly decreased(all P<0.001).Compared with the model group,the fecal water content of the Changzhou Tongbian Formula high-,midium-,low-dose groups and the lactulose group were significantly increased(P<0.001,P<0.01).The small intestinal propu

关 键 词：畅舟通便方 TRPV4 功能性便秘 抑郁 神经可塑性 BDNF/TrkB/CREB信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]