论文部分内容阅读
目的:观察当归对血虚便秘模型小鼠的治疗作用及对小鼠结肠水通道蛋白4(AQP4)表达的影响。方法:用乙酰苯肼、环磷酰胺和复方地芬诺酯联合复制血虚便秘小鼠模型,在观察当归对血虚便秘小鼠的血虚便秘体征、体重、血液学指标(RBC、WBC、HGB和HCT)、粪便排出时间等血虚便秘指标基础上,采用免疫组织化学技术(IHC)、实时荧光定量聚合酶链反应技术(RT-PCR)和蛋白质免疫印迹技术(WB)检测小鼠结肠AQP4的表达,采用酶联免疫技术(ELISA)检测小鼠结肠PLC-IP3-CaM途径中各因子的表达量,探讨当归润肠通便作用及其机制。结果:与血虚便秘组比较,3个剂量(16.67、8.33、4.17g/kg)当归水煎液均能改善模型小鼠血虚便秘证候,均能升高血虚便秘小鼠外周血RBC、WBC、HGB和HCT、缩短排便时间;3个剂量当归水煎液均可显著降低血虚便秘小鼠结肠AQP4和AQP4mRNA的表达;16.67g/kg的当归水煎液组小鼠结肠PLC显著降低;3个剂量当归水煎液组小鼠结肠IP3均有降低趋势,但差异无统计学意义;3个剂量当归水煎液组小鼠结肠CaM均显著降低。结论:当归对血虚便秘模型小鼠具有一定的治疗作用,其作用机制可能与调节结肠PLC-IP3-CaM信号通路从而下调结肠AQP4和AQP4mRNA的表达,减少结肠水分重吸收有关。
OBJECTIVE: To observe the therapeutic effect of Angelica sinensis on mice with constipation-deficient model and the effect on the expression of aquaporin-4 (AQP4) in mice. Methods: Aceticacid hydrazide, cyclophosphamide and diphenoxylate were combined to replicate the blood-deficient constipation mouse model. The incidence of constipation, weight, hematological parameters (RBC, WBC, HGB and HCT), excretion time and other constipation-related indicators of blood deficiency on the basis of the use of immunohistochemistry (IHC), real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and Western blotting (WB) The expression of AQP4 was detected by enzyme-linked immunosorbent assay (ELISA), and the effect of angelica colon and intestine on constipation was investigated. Results: Compared with the blood-stasis constipation group, all three doses (16.67,8.33 and 4.17g / kg) of Danggui Decoction could improve the syndrome of blood deficiency and constipation in model mice, , WBC, HGB and HCT to shorten the time of defecation. The three doses of Angelica decoction could significantly reduce the expression of AQP4 and AQP4 mRNA in the colon of blood-deficient constipation mice. The colon PLC of 16.67g / kg Angelica decoction decreased significantly ; 3 doses of Angelica decoction group mice colon IP3 decreased, but the difference was not statistically significant; 3 doses of Angelica decoction group mice colon CaM were significantly lower. Conclusion: Angelica sinensis has a certain therapeutic effect on mice with blood deficiency constipation. Its mechanism may be related to the regulation of colon PLC-IP3-CaM signaling pathway to down-regulate the expression of AQP4 and AQP4 mRNA and decrease the reabsorption of colon water.