大黄对洛哌丁胺便秘模型大鼠结肠水通道蛋白3表达及肠动力的作用研究  

Influence of Rhubarb on Expression of Aquaporin 3 in Colon and Intestinal Motility in Loperamide-induced Constipated Rats

作　　者：占煜 申旭龙 徐红[1] 唐学贵[2] 

机构地区：[1]成都市中西医结合医院,四川成都610041 [2]川北医学院附属医院,四川南充637000

出　　处：《中华中医药学刊》2016年第10期2315-2320,I0001,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81573990);四川省教育厅计划项目(11ZZ025);川北医学院科研发展计划项目(CBY13-A-ZD08)

摘　　要：目的:观察大黄对洛哌丁胺便秘模型大鼠肠道动力以及结肠AQP3功能和表达的干预作用,探讨大黄发挥通便效应的作用机制。方法:50只Wistar大鼠随机分为5组,分别是空白组(A)、模型组(B)、大黄+消炎痛组(C)、大黄+安慰剂组(D)、消炎痛+安慰剂组(E)。采用洛哌丁胺(1.5mg·kg^(-1)·d^(-1))给予B、C、D、E组大鼠灌胃7 d,建立便秘模型,处死A、B组大鼠比较一般情况、粪便含水量、炭末推进率以验证模型。随后,用生大黄粉(5 g·kg^(-1))混悬液予C、D组大鼠灌胃,E组则采用同等剂量生理盐水。C、E组大鼠灌胃前以消炎痛(10mg·kg^(-1))预处理,D组则使用生理盐水代替。5 h后处死C、D、E组大鼠。所有大鼠处死前均以10%活性炭悬液灌胃,测定炭末推进率。采用RT-PCR、免疫组织化学法、ELISA检测大鼠近端结肠组织中AQP3表达情况以及PGE2、VIP的活性水平。结果:1造模后大鼠的体质量增加、结肠存留粪便增多、粪便含水量降低、肠道传输时间延长(均P<0.05)。2大黄干预便秘大鼠,可加快肠道传输、增加粪便含水量、降低VIP浓度、升高PGE2含量、下调AQP3 mRNA及蛋白表达水平(均P<0.05)。3消炎痛能够显著抑制大黄的通便效应,减少粪便含水量、降低PGE2浓度、上调AQP3 mRNA及蛋白表达水平(均P<0.05),但对肠道传输和VIP无明显影响(均P>0.05)。结论:1通过刺激结肠产生PGE2,快速下调上皮细胞质膜AQP3表达水平,抑制结肠吸收水分,可能是大黄治疗便秘的作用机制之一。2大黄可增强便秘大鼠的肠传输功能,可能与其降低结肠VIP水平,促进肠道蠕动有关。Objective： The aim of this study was to examine the role of AQP3 and intestinal motility in the intervention of rhubarb for loperamide - induced constipated rats so as to explore the mechanism of laxative effect of rhubarb. Methods ： Thirty Wistar rats were randomly divided into five groups, including control group （ A）, model group （ B ）, rhubarb plus indomethacin group（ C）, rhubarb plus placebo group（D） and placebo plus indomethacin group（E）. Afterwards, it was established for constipation model by oral administration of loperamide （ 1.5 mg · kg- 1 · d-1） to rats in the B, C, D and E groups during seven days. Subsequently,rats in the A and B groups were killed to validate the constipation model by com- paring physical condition, fecal water content and charcoal - powder propulsion rate. After that, suspension of Rhez Radix et Rhizoma（5 g·kg-1 ） was administered orally to rats in the C and D groups,which was replaced by saline with the same dose to rats in E group. Additionally,indomethacin（ 10 mg·kg-1） or saline was administered intraperitoneally to rats in C, D and E groups（ saline only for D group） fifteen minutes before the lavage. Rats were autopsied after injecting administration of 10% active charcoal suspension for detecting charcoal - powder propulsion rate and the colons were removed five hours after treatment. RT - PCR, Immunohistochemistry and ELISA were used to examine the expression of AQP3 as well as concentrations of PGE2 and VIP in the proximal colon. Results ：（1）There was significant increase in body weight ,feces in the colon and intestinal transit time whereas fecal water content was remarkable reduced in model rats（ P〈 0.05 ）. （2）It was observed that rhubarb could significantly accelerate the intestinal transit, raise fecal water content, decrease the concentration of VIP, increase the concentration of PGE2 and down - regulate the expression levels of AQP3 in constipated rats （ P 〈 0.05 ）. （3）When pretreated with indometha

关 键 词：大黄 水通道蛋白3 前列腺素E2 血管活性肠肽 洛哌丁胺便秘模型 通便机制 结肠 

分 类 号：R285.5[医药卫生—中药学]