车前子对腹泻大鼠炎性因子和结肠组织AQP4 mRNA和蛋白表达的影响  

Effect of Plantaginis Semen on Inflammatory Cytokines and mRNA and Protein Expressions of AQP4 in Colon Tissue of Diarrhea Rats

作　　者：王月 彪雅宁 储心乔 张纳博 张睦清[2] 王茜 韩雪 张一昕 WANG Yue;BIAO Ya-ning;CHU Xin-qiao;ZHANG Na-bo;ZHANG Mu-qing;WANG Xi;HAN Xue;ZHANG Yi-xin(School of Pharmacy,Hebei University of Chinese Medicine,Application Technology Research Center of Traditional Chinese Medicine(TCM)Prescription in University of Hebei Province,Shijiazhuang 050200,China;Hebei Hospital of TCM,Shijiazhuang 050011,China)

机构地区：[1]河北中医学院药学院,河北省高校中药组方制剂应用技术研究中心,石家庄050200 [2]河北省中医院,石家庄050011

出　　处：《中国实验方剂学杂志》2020年第20期52-58,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省中医药管理局科研计划项目(2018210);中央财政公共卫生专项“2018年中药资源普查”项目(Z135080000022);国家级大学生创新创业训练计划项目(201714432001)。

摘　　要：目的:观察车前子对腹泻大鼠血清中炎性因子和结肠组织水通道蛋白4(AQP4)mRNA和蛋白表达的影响,探讨其治疗腹泻的作用机制。方法:采用番泻叶药液灌胃法复制大鼠腹泻模型。将60只SD雄性大鼠随机分为正常组、模型组、氢氯噻嗪组(9 mg·kg^-1)和车前子低、中、高剂量组(0.95,1.9,3.8 g·kg^-1)。除正常组外,其余各组均于每日上午灌胃番泻叶药液(20 mL·kg^-1),正常组灌胃等量的蒸馏水;每日下午各治疗组灌服相应药物,正常组和模型组灌服等量蒸馏水,连续14 d。实验结束后,根据粪便性状和大便次数,统计对比各组大鼠的稀便率、平均稀便级和腹泻指数,采集血清和结肠组织标本,检测血清中肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6)和C反应蛋白(CRP)的含量;苏木素-伊红(HE)染色观察结肠黏膜病理形态学变化;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测结肠组织中AQP4 mRNA和蛋白的表达情况。结果:与正常组比较,模型组稀便率、平均稀便级和腹泻指数显著升高(P<0.01),结肠黏膜表皮细胞出现凋亡坏死,固有层毛细血管扩张充血明显,少量中性粒细胞浸润,血清中TNF-α,IL-6和CRP的含量明显升高(P<0.05,P<0.01),AQP4 mRNA和蛋白的表达均明显下降(P<0.01);与模型组比较,车前子各剂量组大鼠的稀便率、平均稀便级和腹泻指数均显著降低(P<0.01),结肠黏膜表皮细胞凋亡坏死、毛细血管扩张充血和中性粒细胞浸润现象明显改善,血清中TNF-α和CRP的含量降低(P<0.05,P<0.01),AQP4 mRNA和蛋白的表达水平明显升高(P<0.05,P<0.01)。结论:车前子具有较好的止泻作用,其机制可能是抑制炎症反应、修复结肠黏膜病理损伤,上调AQP4的表达,调节水液代谢。Objective:To observe Plantaginis Semen’s mechanism in treating diarrhea by observing the effect on inflammatory factors in serum and mRNA and protein expressions of aquaporin4(AQP4)in colon tissue of diarrhea rats. Method: Senne Folium was orally administered to duplicate diarrhea rats. Sixty male SD rats were randomly divided into normal group,model group,hydrochlorothiazide group(9 mg·kg^-1),and low,middle,and high-dose Plantaginis Semen groups(0. 95,1. 9,3. 8 g·kg^-1). Senne Folium(20 mL·kg^-1)was intragastrically administered in 5 groups in the morning,except for normal group that was orally given the same dose of distilled water. In the afternoon,each treatment group was orally given the corresponding drugs,while normal group and model group were orally given the same dose of distilled water. The loose stool rate,average degree of loose stool,and diarrhea index were compared according to fecal traits and stool times after 14 days of treatment. The serum and colon tissue were collected to detect the contents of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and C-reactive protein(CRP)in serum. Hematoxylin-eosin(HE)staining was used to observe the pathological morphological changes of colon tissue,and quantiative Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the mRNA and protein expressions of AQP4 in colon tissue. Result: In the model group,the loose stool rate,average degree of loose stool,and diarrhea index were significantly increased(P<0. 01),apoptosis and necrosis were observed in the epidermal cells of colonic mucosa,telangiectasia and congestion in lamina propria were obvious,and a few neutrophils were infiltrated,and the contents of TNF-α,IL-6 and CRP in serum increased(P<0. 05,P<0. 01),the mRNA and protein expressions of AQP4 significantly decreased(P<0. 01). Compared with the model group,the loose stool rate,average degree of loose stool,and diarrhea index were significantly decreased in low,middle,and high-dose Plantaginis

关 键 词：腹泻 车前子 肿瘤坏死因子-α(TNF-α) 白细胞介素-6(IL-6) C反应蛋白(CRP) 水通道蛋白4(AQP4) 

分 类 号：R2-0[医药卫生—中医学] R22