基于VIP-cAMP-PKA-AQP3信号通路探讨枳实-白芍药对调控C-IBS的作用及机制  

Effect and mechanism of couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba on regulation of C-IBS based on VIP-cAMP-PKA-AQP3 signaling pathway

作　　者：辛丹 梁晓东[1] 滕佳林[1] 张亚楠[1] 马晓麒 唐迎雪[1] XIN Dan;LIANG Xiao-dong;TENG Jia-lin;ZHANG Ya-nan;MA Xiao-qi;TANG Ying-xue(Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学,济南250355

出　　处：《中华中医药杂志》2022年第11期6461-6465,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：山东省重点产业关键技术项目(No.2016CYJS08A01-3,No.2016CYJS08A01-4);山东省重点研发计划(No.2016ZDJS07A12)。

摘　　要：目的:研究不同配伍比例的枳实-白芍药对对便秘型肠易激综合征(C-IBS)大鼠的影响,探讨其对C-IBS的作用及机制。方法:取SPF级Wistar成年雄性大鼠60只,随机留取10只大鼠作为空白组。其余50只大鼠为实验组,均采用冰0.9%氯化钠溶液灌胃的方法制备C-IBS模型,造模成功后将50只实验组大鼠随机分为模型对照组、匹维溴铵组、枳实-白芍药对1∶1组、枳实-白芍药对1∶2组、枳实-白芍药对2∶1组,每组10只。经过14 d治疗后,计算粪便含水量,ELISA法检测血清血管活性肠肽(VIP)、环磷酸腺苷(cAMP)、蛋白激酶A(PKA)、水通道蛋白3(AQP3)的含量,荧光定量PCR检测结肠组织VIP、PKA、AQP3 mRNA的表达,Western Blot检测结肠组织cAMP、PKA、AQP3蛋白的表达。结果:与模型对照组比较,枳实-白芍药对1∶1组、枳实-白芍药对1∶2组、枳实-白芍药对2∶1组均可升高粪便含水量(P<0.05),枳实-白芍药对2∶1组可显著升高血清VIP、cAMP、PKA、AQP3含量(P<0.01),枳实-白芍药对1∶1组可升高血清VIP含量(P<0.05),枳实-白芍药对1∶2组可显著升高血清VIP、cAMP、AQP3含量(P<0.01,P<0.05),枳实-白芍药对1∶1组、枳实-白芍药对1∶2组、枳实-白芍药对2∶1组均可显著升高结肠组织VIP、PKA、AQP3 mRNA和cAMP、PKA、AQP3蛋白表达水平(P<0.05,P<0.01),且各组之间差异无统计学意义,说明枳实和白芍配伍时以经典名方枳实芍药散的原方剂量即可。结论:枳实-白芍药对配伍比例为1∶1时即可发挥治疗C-IBS的作用,其机制可能与调控VIP-cAMP-PKA-AQP3信号通路有关。Objective:To study the effect of different compatibility proportion of couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba on constipation-predominant irritable bowel syndrome(C-IBS)model rats,and to explore its effect and mechanism on C-IBS.Methods:A total of 60 adult male SPF Wistar rats were selected,and 10 rats were randomly selected as normal group.The other 50 rats served as the experimental group,and the C-IBS model was established by intragastric administration of ice 0.9%sodium chloride solution.After the model was prepared successfully,the 50 rats in the experimental group were randomly divided into model control group,pinaverium bromide group,couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 1∶1 group,couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 1∶2 group and couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 2∶1 group,with 10 rats in each group.After 14 days of treatment,the fecal water content was calculated,the contents of VIP,cAMP,PKA and AQP3 in serum were detected by ELISA,the mRNA expressions of VIP,PKA and AQP3 in colon tissues were detected by fluorescence quantitative PCR,and the protein expressions of cAMP,PKA and AQP3 in colon tissues were detected by Western Blot.Results:Compared with the model control group,groups of 1∶1,1∶2 and 2∶1 of couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba could increase fecal water content(P<0.05),couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 2∶1 group could increase contents of VIP,cAMP,PKA and AQP3 in serum(P<0.01),couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 1∶1 group could increase serum VIP content(P<0.05),couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba 1∶2 group could increase contents of VIP,cAMP and AQP3 in serum(P<0.01,P<0.05),groups of 1∶1,1∶2 and 2∶1of couplet medicine of Aurantii Fructus Immaturus-Paeoniae Radix Alba could increase mRNA levels of VIP,PKA and AQP3 and protein ex

关 键 词：枳实 白芍 药对 便秘型肠易激综合征 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 机制 经方 配伍 

分 类 号：R285.5[医药卫生—中药学]