摘要
目的检测TNF-α结合肽和TNFR封闭肽对TNBS诱导的大鼠溃疡性结肠炎的治疗作用。方法以TNBS诱导的大鼠溃疡性结肠炎为模型,联合给予TNF-α结合肽和TNFR封闭肽[2.5mg/(kg·次)]处理,同时设各种对照;观察大鼠的症状、体重、结肠组织病理改变等;采用一般的生物化学检测方法检测大鼠血清和结肠组织中NO含量、活性氧以及结肠组织中MP0活性等炎性介质;采用RT-PCR技术检测大鼠腹腔MIL-1β和IL-8等mRNA水平的变化;采用免疫组织化学SP法检测大鼠结肠组织中TNF-α蛋白质表达水平的改变。结果TNF结合肽和TNFR封闭肽联合多肽组大鼠的体重、结肠组织形态学评分、组织病理学评分及血清和结肠组织中NO含量、MP0活性等指标均显著低于模型组及无关肽组(P<0.01);其腹腔M的IL-1β和IL-8 mRNA水平和结肠组织中TNF-α蛋白的表达水平及阳性细胞率也显著低于显著低于模型组及无关肽组(P<0.01)。结论TNF结合肽和TNFR封闭肽联用可显著减轻TNBS诱导大鼠实验性溃疡性结肠炎的病理损伤,改善症状,有效抑制大鼠血清和结肠组织中NO含量以及结肠组织中MP0活性的活性,抑制大鼠腹腔巨噬细胞中Il-1β、IL-8的mRNA转录和结肠组织中TNF-α蛋白的表达,对TNBS诱导的大鼠溃疡性结肠炎具有治疗作用。本研究为进一步研制和开发拮抗TNF-α的新型肽类药物提供了重要的实验依据。
Objective To examine the therapeutic effects of TNF-α binding peptide （TBP） and TNFR blocking peptide （TRBP） on TNBS-induced colitis in rats. Methods The combination of TBP and TRBP （2.5 mg/kg, once a day） were administered orally to treat the rats with TNBS-induced colitis, while the control animals simultaneously were treated with sodium saline or irrelevant peptide. The macroscopic score and histological score of colonic damage were assessed; the myelopemxidase （MPO） activity and nitric oxide （NO） production in serum and the colonic tissue were determined; the mRNA expressions of proinflammatory cytokines, such as IL-1β and IL-8, in the peritoneal macmphages were assayed by RT-PCR; protein expression of TNF-α in the colonic tissue was also investigated by SP immtmohistochemistry assay. Results Macroscopic score, histological score, MPO activity, and NO production in rats treated with the combination of TBP and TRBP were significantly lower than those in groups of irrelavant peptide and sodium saline （ P 〈 0.01 ）. The protein expression of TNF-α and the mRNA expressions of IL-1β and IL-8 in rats treated with the combination of TBP and TRBP were also down-regulated （ P 〈 0.01 ）, compared with that in rats treated with the irrelevant peptide or sodium saline; the number of TNF-α positive cells in peptide-treatment group was larger than that in the two control group too （ P 〈 0.01 ）. Conclusion TNF-α binding peptide and TNFR blocking peptide obviously improve the symptoms of the TNBS-induced colitis in rats and alleviate the colonic pathological damage. This research will provide an experimental basis for the clinical treatment of the inflammatory disease.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第6期663-667,共5页
Immunological Journal
基金
国家"863"高技术研究发展计划资助项目(2004AA215162)
