硝菔通结方对功能性便秘大鼠结肠组织中VIP-cAMP-PKA-AQP3信号通路的影响

目的:探讨硝菔通结方对功能性便秘大鼠结肠组织中VIP-c AMP-PKA-AQP3信号通路的影响。方法:SD雄性大鼠150只,随机分为5组,分别为正常组、模型组、硝菔通结方高、中、低剂量组(380,190,95 g·kg-1),每组30只,除正常组外采用复方地芬诺酯ig法建立SD大鼠功能性便秘模型,给予不同剂量硝菔通结方ig,共给药4周。检测大鼠首粒黑边排出时间及粪便含水率;采用实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白质免疫印迹(Western blot)分别检测治疗1,2,3周不同时间点大鼠结肠组织中血管活性肠肽(VIP),环磷酸腺苷(c AMP),蛋白激酶A(PKA),水通道蛋白3(AQP3)mRNA及蛋白质的表达变化。结果:与正常组比较,模型组大鼠首粒黑便排出时间显著延长,粪便含水率显著降低(P0.01),结肠组织中VIP,c AMP,PKA,AQP3的mRNA和蛋白表达水平明显降低(P0.05,P0.01);与模型组比较,硝菔通结方各剂量组大鼠首粒黑便排出时间显著缩短,粪便含水率明显增加(P0.05,P0.01),结肠组织中VIP,c AMP,PKA,AQP3的mRNA和蛋白表达水平均有升高趋势,且高、中剂量组较低剂量组升高更显著,给药后第2,3周较第1周的指标变化更显著(P0.05,P0.01)。结论:硝菔通结方能够调节胃肠动力和改善肠道水液代谢治疗功能性便秘,其机制可能是通过干预VIP-c AMP-PKAAQP3通路来实现,并且具有一定的时间和剂量依赖性。

Effect of Xiaofu Tongjie Recipe on VIP-cAMP-PKA-AQP3 Signaling Pathway in Colon Tissues of Rats with Functional Constipation

Objective: To explore the effect of Xiaofu Tongjie recipe on VIP-cAMP-PKA-AQP3 signaling pathway in colon tissues of rats with functional constipation in rats. Method: Totally 150 SD male rats were randomly divided into normal group, model group, Xiaofu Tongjie recipe high dose, middle dose and low dose groups (380, 190, 95 g·kg^-1), n=30 in each group. Compound diphenoxylate was applied by ig to induce functional constipation SD rat models, and then different doses of Xiaofu Tongjie recipe were given by ig administration for 4 weeks. The excretion time of first black defecating feces and moisture content in the feces were detected. The Real-time PCR and Western bolt were used respectively to detect the mRNA and protein expression levels of vasoactive intestinal peptide (VIP), cyclic adenosine monophosphate (cAMP), protein kinase A (PKA) and aquaporin 3 (AQP3) at different time points after treatment of 1, 2, 3 weeks. Result: As compared with the normal group, the excretion time of first black defecating feces was significantly prolonged in model group; moisture content in the feces was significantly reduced (P<0.01); the mRNA and protein expression levels of VIP, cAMP, PKA and AQP3 in colon tissues were significantly reduced (P<0.05, P<0.01). As compared with the model group, the excretion time of first black defecating feces was significantly shortened in various Xiaofu Tongjie recipe groups, and the moisture content in the feces was significantly increased (P<0.05, P<0.01); the mRNA and protein expression levels of VIP, cAMP, PKA and AQP3 in colon tissues were increased, and the increase was more obvious in high and middle dose groups; the changes of the indexes at week 2 and week 3 were more obvious than those of week 1 (P<0.05, P<0.01). Conclusion: Xiaofu Tongjie recipe can regulate the intestinal peristalsis and intestinal fluid metabolism and cure the functional constipation in a time and dose-dependent manner, which may be achieved by intervention on VIP-cAMP-PKA-AQP3 signaling pathway.