基于VIP-cAMP-PKA-AQP3信号通路探讨益气润肠方治疗气虚型便秘大鼠的作用机制  

Action Mechanism of Yiqi Runchang Formula in Treatment of Qi-Deficiency Constipation Rats Based on VIP-cAMP-PKA-AQP3 Signaling Pathway

作　　者：熊润秋 高月[1] 陈鲁[2] XIONG Runqiu;GAO Yue;CHEN Lu(Shandong University of Traditional Chinese Medicine,Jinan 250014,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学,山东济南250014 [2]山东中医药大学附属医院,山东济南250014

出　　处：《中医药信息》2023年第9期10-15,共6页Information on Traditional Chinese Medicine

基　　金：山东省中医药科技发展计划项目(2017-073)。

摘　　要：目的:基于VIP-cAMP-PKA-AQP3信号通路探讨益气润肠方对气虚便秘模型大鼠作用及其机制。方法:选取48只SD大鼠随机分为空白组、模型组、乳果糖组、益气润肠方低剂量组、益气润肠方中剂量组和益气润肠方高剂量组,每组8只。空白组大鼠正常饮食,每日给予生理盐水灌胃,其余大鼠给予洛哌丁胺混悬液3 mg/(kg·d)灌胃加饥饱饮食2周制作气虚便秘模型。造模成功后,空白组及模型组每日给予生理盐水灌胃,乳果糖组给予乳果糖口服溶液1.67 g/(kg·d)灌胃,益气润肠方高、中、低剂量组分别给予益气润肠方17.5、8.75、4.375 g/(kg·d)灌胃,连续2周。14 d后称量大鼠粪便湿重、粪便干重并计算粪便含水率;留取结肠组织,采用苏木精伊红染色法观察结肠病理变化,qPCR检测大鼠结肠血管活性肠肽(VIP)、环磷酸腺苷(cAMP)、蛋白激酶A(PKA)、水通道蛋白3(AQP3)mRNA表达水平,Western blot检测大鼠VIP、AQP3蛋白表达水平。结果:与空白组比较,模型组大鼠体质量明显降低(P<0.01),粪便湿重及粒数明显减少(P<0.01);与模型组比较,益气润肠方低、中、高剂量组和乳果糖组体质量明显增高(P<0.01,P<0.05),粪便湿重增加、粪便粒数增多(P<0.05,P<0.01),但粪便含水率无统计学意义(P>0.05)。与乳果糖组比较,益气润肠方中、高剂量组粪便湿重明显增加、粪便粒数显著增多(P<0.05)。与空白组比较,模型组VIP、cAMP、PKA、AQP3 mRNA表达水平明显降低(P<0.01,P<0.05);与模型组比较,乳果糖组及益气润肠方各剂量组VIP、cAMP、PKA、AQP3 mRNA表达水平明显升高(P<0.05,P<0.01),益气润肠方高剂量组效果最显著(P<0.01),且优于乳果糖组(P<0.05)。与模型组比较,益气润肠方各剂量组VIP、AQP3的蛋白表达水平升高,且呈剂量依赖性(P<0.05)。益气润肠方可以明显改善咯派丁胺造成的便秘大鼠的肠道病理形态损害。结论:益气润肠方可通过调控VIP-cAMP-PKA-AQP3通路Objective:To investigate the effect and mechanism of Yiqi Runchang Formula in treating model rats with Qi-deficiency constipation.Methods:48 SD rats were randomly divided into the blank group,the model group,the lactutose group(1.67 g/kg·d),the Yiqi Runchang Formula groups of low-dose(4.375 g/kg·d),medium-dose(8.75 g/kg·d)and high-dose(17.5 g/kg·d).The model of Qi-deficiency constipation was established by loperamide suspension(3 mg/kg·d)gavage plus starvation and satiety diet for 2 weeks.After 14 days of corresponding intervention,wet fecal stool weight and dry fecal stool weight were measured to calculate water content of fecal stool.Colon tissues were collected and HE staining was used to observe the pathological changes of colon.mRNA expressions of vasoactive intestinal peptide(VIP),cyclic adenosine phosphate(cAMP),protein kinase A(PKA)and aquaporin 3(AQP3)were detected by qPCR.The protein expressions of VIP and AQP3 were detected by Western blot.Results:Compared with those in the blank group,the body mass was significantly decreased(P<0.01),the weight of wet fecal stool and the fecal pellet number were significantly reduced in the model group(P<0.01).Compared with those in the model group,the body mass was significantly increased(P<0.01,P<0.05),the weight of wet fecal stool and the fecal pellet number were significantly increased in the lactulose group and the Yiqi Runchang Formula groups of low-dose,medium-dose and high-dose(P<0.05,P<0.01);but water content of fecal stool showed no statistical difference(P>0.05).Compared with those in the lactulose group,the weight of wet fecal stool and the fecal pellet number were significantly increased in the Yiqi Runchang Formula groups of medium-dose and high-dose(P<0.05).Compared with those in the blank group,mRNA expressions of VIP,cAMP,PKA and AQP3 were significantly decreased in the model group(P<0.01,P<0.05).Compared with those in the model group,mRNA expressions of VIP,cAMP,PKA and AQP3 were significantly increased in the lactulose group and the Yiqi Runcha

关 键 词：功能性便秘 血管活性肠肽-环磷酸腺苷-蛋白激酶A-水通道蛋白3信号通路 洛哌丁胺 益气润肠方 

分 类 号：R285.5[医药卫生—中药学]