消积通便方对便秘小鼠VIP-cAMP-PKA信号通路的影响  

Effects of Xiaoji Tongbian Decoction on VIP-cAMP-PKA signal pathway in mice with constipation

作　　者：胡钰[1,2] 候淑婷 HU Yu;HOU Shuting(Nantong Affiliated Hospital of Nanjing University of Chinese Medicine, Nantong 226001, Jiangsu, China;The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, Jiangsu, China;Jiangsu Key Laboratory of Pediatric Respiratory Disease (Chinese Medicine), Nanjing 210023, Jiangsu, China)

机构地区：[1]南京中医药大学南通附属医院,江苏南通226001 [2]南京中医药大学附属医院,江苏南京210029 [3]江苏省儿童呼吸疾病(中医药)重点实验室,江苏南京210023

出　　处：《现代中西医结合杂志》2022年第4期485-490,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：南通市卫生和计划生育委员会科研课题(MB2020015);南通市科技计划项目(JCZ20199)。

摘　　要：目的探讨消积通便方治疗便秘的作用机制。方法将64只SPF级雄性ICR小鼠随机分为空白组、模型组、消积通便方低剂量组、消积通便方中剂量组、消积通便方高剂量组、莫沙必利组。空白组给予生理盐水灌胃,其余各组给予洛哌丁胺10 mg/kg灌胃,均每日2次,共14 d。消积通便方低、中、高剂量组以及莫沙必利组小鼠均于第8天开始灌胃相应药物,正常组和模型组灌胃等量的生理盐水,均连续7 d。观察各组小鼠一般情况,记录实验第15天胃排空率及肠道推进率,采用ELISA法测定血清中血管活性肠肽(VIP)和诱导型一氧化氮合酶(iNOS)水平,HE染色观察结肠组织黏膜情况,实时荧光定量PCR法检测结肠组织中环磷酸腺苷(cAMP)、蛋白激酶A(PKA)mRNA表达情况。结果空白组小鼠一般状态良好,体重逐步增加;模型组小鼠有不同程度易激惹或倦怠,精神不振,反应迟钝,进食量及饮水量减少;消积通便方各组及莫沙必利组小鼠精神状态及活动改善,进食、饮水量渐增。与空白组比较,模型组小鼠胃排空率及血清VIP、iNOS水平均明显增高(P均<0.05),肠道推进率及结肠组织中PKA mRNA、cAMP mRNA表达量均明显降低(P均<0.05);HE染色显示结肠黏膜固有层及黏膜下层可见炎性细胞浸润,肌层明显变薄。与模型组比较,消积通便方中、高剂量组及莫沙必利组小鼠胃排空率及血清VIP、iNOS水平均明显降低(P均<0.05),消积通便方高剂量组和莫沙必利组小鼠肠道推进率及结肠组织中PKA mRNA、cAMP mRNA表达量均明显升高(P均<0.05);HE染色显示消积通便方中、高剂量组及莫沙必利组小鼠结肠组织损伤明显减轻。结论消积通便方可能通过下调VIP、iNOS水平,激活cAMP-PKA通路,从而调节肠道平滑肌的活动,促进肠蠕动,缓解便秘症状。Objective It is to explore the mechanism of Xiaoji Tongbian(XJTB)Decoction in the treatment of constipation.Methods 64 male ICR mice of SPF grade were randomly divided into blank group,model group,XJTB low-dose group,medium-dose group,high-dose group and mosapride group.The blank group was given normal saline by gavage.The other groups were given loperamide 10 mg/kg by gavage,twice a day,totally for 14 days.The mice in XJTB low,medium and high groups and mosapride group were given corresponding drugs intragastrically on the 8 th day,the mice in the normal group and the model group were given the same amount of normal saline by intragastric administration,all treated for 7 consecutive days.The general condition of the mice was observed,the gastric emptying rate and intestinal propelling rate of the mice were calculated,the levels of vasoactive intestinal peptide(VIP)and inducible nitric oxide synthase(iNOS)in serum were determined by ELISA assay,and the pathological manifestations of intestinal mucosal tissue was observed by HE staining,the mRNA expressions of Cyclic adenosine monophosphate(cAMP),protein kinase A(PKA)were detected by RT-PCR.Results The mice in the blank group were in good condition and their body weight increased gradually.The mice in the model group had different degrees of irritability or tiredness,inactivity,retarded reaction,reduced food and water intake.The mental state and activity of mice in every XJTB group and mosapride group were improved,and the intake of food and water was increased.Compared with the blank group,the gastric emptying rate and serum VIP and iNOS levels of mice in the model group were significantly increased(all P<0.05),while the intestinal propulsion rate and the expressions of PKA mRNA and cAMP mRNA in colon tissue were significantly decreased(all P<0.05);HE staining showed that inflammatory cells were infiltrated in the lamina propria and submucosa of the colonic mucosa,and the muscular layer was obviously thinned.Compared with the model group,the gastric emptying rate

关 键 词：消积通便方 便秘 血管活性肠肽 诱导型一氧化氮合酶 环磷酸腺苷-蛋白激酶A通路 

分 类 号：R-332[医药卫生]