粪便移植对艰难梭菌引起的大鼠伪膜性肠炎的作用机制研究

Therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium diffi-cile-associated pseudomembranous colitis

目的：研究粪便移植对艰难梭菌引起的大鼠伪膜性肠炎的治疗作用和机制。方法40只SD大鼠分为健康对照组、模型组、粪便移植组和万古霉素组,后3组皮下注射克林霉素（10 mg/kg）,24 h后用（产毒）艰难梭菌（105 CFU/ml）灌肠造模,造模1 d后分别用粪便悬液和万古霉素治疗。末次给药后禁食1 d,股动脉取血,对K、Na、血清白蛋白（ ALB）、白细胞计数（ WBC）、中性粒细胞百分率（ N%）、C-反应蛋白（ CRP）、IL-1β、IL-10、IL-12和IL-17水平进行检测,取结肠行病理学检查。结果通过皮下注射克林霉素加（产毒）艰难梭菌诱导成功建立大鼠伪膜性肠炎模型,模型组大鼠体重和结肠长度显著降低,结肠湿质量指数显著升高（P〈0.05）。与模型组比较,粪便移植组K和ALB明显升高（P〈0.05）,WBC、N%和CRP明显降低（P〈0.05）,IL-1β和IL-17明显降低（P〈0.05）, IL-10和 IL-10/IL-12含量升高（P〈0.05）。结论粪便移植法可有效治疗伪膜性肠炎,机制与其影响血清炎症因子水平有关。

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcuta-neously injected with clindamycin phosphate （10 mg）, followed by treatment with toxin producing Clostridi-um difficile （ACTT43255） enema 24 hours later. The rats in fecal transplant treatment group and vancomy-cin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion （ K） , sodium ion （ Na） , albumin （ ALB） , white blood cells （ WBC） , C-reaction protein （ CRP） , interleu-kin-1β （ IL-1β） , interleukin-10 （ IL-10 ） , interleukin-12 （ IL-12 ） and interleukin-17 （ IL-17 ） as well as the percentage of neutrophils （ N%） in serum samples were detected. The colon tissue samples were collect-ed for pathology examination. Results The rat model of pseudomembranous colitis was successfully estab-lished by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile （ACTT43255） enema. The signs of intestinal inflammation including serious weight loss, remarkably short-ened colon length and significantly increased colon wet weight index were observed in rats from the model group （P〈0. 05）. Compared with the rats from model group, the rats received fecal transplant showed sig-nificantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 （P〈0. 05）. Conclusion Fecal transplantation was proved to be an effective ap-proach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors.