目的探讨活动期溃疡性结肠炎(ulcerative colitis, UC)患者外周血中多种细胞因子的变化以及不同浓度的黄芩苷的干预作用。方法应用实时定量PCR法(Q-PCR)检测UC患者外周血单个核细胞IL4R、 IL6R、 IL23R、 RORC的表达,采用酶联免疫吸附法(ELISA )检测血清干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、转化生长因子-β1(TGF-β1)水平及体外黄芩苷干预下上述指标的变化情况。结果通过Q-PCR检测发现UC患者外周血IL4R、 IL6R、 IL23R、 RORC基因表达较肠易激综合征(IBS-D)组和健康对照组均有不同程度的增高,在体外不同浓度的黄芩苷的干预下,均有不同程度的降低,以20μmol/L和40μmol/L对上述指标的降低作用更明显。 ELISA检测血清中细胞因子, UC组IFN-γ、 IL-5、 IL-6与IBS-D组和健康对照组比较均有不同程度的升高,而IL-4、 IL-10及TGF-β较IBS-D组和健康对照组稍降低;20μmol/L和40μmol/L的黄芩苷可引起IFN-γ、IL-5、 IL-6明显降低, IL-4、 IL-10明显升高。结论较高浓度的黄芩苷能够明显抑制RORC、 IL23R的表达,使IFN-γ、IL-5、 IL-6等细胞因子的水平降低和IL-4、 IL-10及TGF-β1水平升高,说明黄芩苷可以调节免疫平衡而缓解溃疡性结肠的炎症反应。
Objective To investigate the cytokines levels in peripheral blood of ulcerative colitis (UC) patients at active stage, and to observe the effect of baicalin at various concentrations on the cytokines. Methods Quantitative polymerase chain reaction (Q-PCR) assay was used for the detection of interleukin 4 receptor (IL4R), IL6R, IL23R and RAR-related orphan receptor C (RORC) expression in single peripheral mononuclear cell of UC patients. Enzyme-linked immunosorbent assay (ELISA) was used to examine the serum levels of interferon gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, and transforming growth factor beta (TGF-β) levels in UC patients, and was applied for in-vitro detection of these indicators after baicalin intervention. Results Q-PCR results showed that IL4R, IL6R, IL23R, RORC gene expression levels in UC patients were increased to various degrees as compared to diarrhea-predominant irritable bowel syndrome (IBS-D) patients and the healthy volunteers, and then the levels were decreased to various degrees after intervention by baicalin at different concentrations, in particular at 20 and 40μmol/L. The results of ELISA showed that serum levels of cytokines of IFN-γ, IL-5, IL-6 in UC patients were increased to various degrees while IL-4, IL-10 and TGF-β1 was decreased as compared to IBS-D patients and the healthy volunteers. Baicalin at the concentrations of 20 and 40 μmol/L had an effect on decreasing IFN-γ, IL-5, IL-6 and on increasing IL-4 and IL-10. Conclusion Higher concentrations of baicalin show obvious effect on inhibiting RORC and IL23R expression, on decreasing IFN-γ, IL-5, IL-6 levels, and on increasing IL-4, IL-10 and TGF-β1 levels, which indicated that the therapeutic mechanism of baicalin in relieving ulcerative colonic inflammation is related with the regulation of immune function.