生、制枳实换用的枳实芍药散对便秘型肠易激综合征大鼠“脑-肠-菌”轴功能的影响  

Effect of Raw and Processed Aurantii Fructus Immaturus Switched to Zhishi Shaoyaosan on Function of Brain-Gut-Microbiota Axis in Rats with Constipation-predominant Irritable Bowel Syndrome

作　　者：余远盼 林桂梅[1,2] 李医名 姚怡 YU Yuanpan;LIN Guimei;LI Yiming;YAO Yi(School of Pharmacy,Liaoning University of Traditional Chinese Medicine(TCM),Dalian 116600,China;TCM Processing Technology Inheritance Base of the State Administration of TCM(Liaoning),Dalian 116600,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]国家中医药管理局中药炮制技术传承基地(辽宁),辽宁大连116600

出　　处：《中国实验方剂学杂志》2024年第9期1-9,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81503248);辽宁省自然科学基金项目(2019-MS-230);辽宁省教育厅基本科研项目(LJKMZ20221314)。

摘　　要：目的:从“脑-肠-菌”轴的角度探讨生、制枳实换用的枳实芍药散改善便秘型肠易激综合征(C-IBS)大鼠的作用机制。方法:将80只大鼠随机分为空白组、模型组、匹维溴铵组(15.625 mg·kg^(-1))、生枳实芍药散组、清炒枳实芍药散组、麸炒枳实芍药散组、炒炭枳实芍药散组、成品枳实芍药散组(3.75 g·kg^(-1)),每组10只。除空白组灌胃常温0.9%氯化钠溶液外,其余各组均灌胃0~4℃冰0.9%氯化钠溶液(2 mL·d^(-1),共14 d)建立C-IBS大鼠模型。连续给药14 d后检测大鼠粪便含水量、小肠推进率;酶联免疫吸附测定法(ELISA)检测5-羟色胺(5-HT)、血管活性肠肽(VIP)、神经肽Y(NPY)、降钙素基因相关肽(CGRP)、P物质(SP)、二胺氧化酶(DAO)、D-乳酸(D-LA)水平;苏木素-伊红(HE)染色观察结肠组织病理损伤变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测结肠组织环磷酸腺苷(cAMP)、蛋白激酶A(PKA)、水通道蛋白3(AQP3)mRNA的表达及蛋白免疫印迹法(Western blot)检测结肠组织VIP和AQP3蛋白的表达;采用气质联用色谱法(GC-MS)检测短链脂肪酸(SCFAs)含量的变化。结果:与空白组比较,模型组大鼠粪便含水量、小肠推进率显著降低(P<0.01);血清中5-HT、VIP、CGRP、SP水平显著上升,NPY水平显著下降(P<0.01);血浆中DAO和D-LA水平显著升高(P<0.01);结肠组织黏膜上皮轻度破损,杯状细胞减少且腔内颗粒明显减少;结肠组织中AQP3、cAMP、PKA mRNA表达水平和AQP3、VIP蛋白表达水平明显下降(P<0.05,P<0.01);粪便中SCFAs总量呈明显下降趋势,其中,乙酸、异丁酸、异戊酸、戊酸、己酸含量显著减少,丙酸、丁酸含量明显上升(P<0.05,P<0.01)。与模型组比较,各给药组均明显提高C-IBS大鼠小肠推进率,改善肠黏膜屏障功能,调整模型大鼠血清中脑肠肽水平(P<0.05,P<0.01),明显升高大鼠结肠组织中AQP3、cAMP、PKA mRNA及VIP、AQP3蛋白的表达水平(P<0.05,P<0.01),除异丁酸外各�Objective:To research the mechanism underlying the effect of raw and processed Aurantii Fructus Immaturus switched to Zhishi Shaoyaosan(ZSS)on constipation-predominant irritable bowel syndrome(C-IBS)rats via the brain-gut-microbiota axis.M ethod:Eighty rats were randomly divided into the blank,model,positive drug(pinaverium bromide,15.625 mg·kg^(-1)),raw ZSS,stir-fried ZSS,bran-fried ZSS,charcoal-fried ZSS and finished ZSS groups(3.75 g·kg^(-1)),with 10 rats in each group.Except for the blank group,which received intragastric administration of 0.9%sodium chloride solution at room temperature,all other groups were administered the ice solution at 0 to 4°C(2 mL·d^(-1),for a total of 14 d)to establish the C-IBS rat model.The fecal water content and the propulsion rate of small intestine were detected after 14 d of continuous drug administration.The levels of 5-hydroxytryptamine(5-HT),vasoactive intestinal peptide(VIP),neuro-peptide Y(NPY),calcitonin gene-related peptide(CGRP),substance P(SP),diamine oxidase(DAO)and D-lactic acid(D-LA)were detected by enzyme linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining was used to observe the changes in colonic pathological injury in each group.The expression levels of cyclic adenosine monophosphate(cAMP),protein kinase A(PKA)and aquaporin-3(AQP3)mRNA in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and the protein expressions of VIP and AQP3 in colon tissues were detected by Western blot.The content of short chain fatty acids(SCFAs)was determined by gas chromatography-mass spectrometry.Result:Compared with the blank group,the fecal water content and intestinal propulsion rate of rat in the model group were significantly decreased(P<0.01),and the levels of 5-HT,VIP,CGRP and SP in serum were significantly increased.Simultaneously,the NPY levels significantly decreased(P<0.01),the levels of DAO and D-LA in plasma were significantly increased(P<0.01),and the mucosal epithelium of colon tissue was slightl

关 键 词：枳实芍药散 枳实 炮制 便秘型肠易激综合征 脑-肠-菌轴 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33