新型查尔酮衍生物MW-9抑制MAPK信号通路治疗小鼠溃疡性结肠炎

MW-9,a chalcones derivative bearing heterocyclic moieties,ameliorates ulcerative colitis via regulating MAPK signaling pathway

目的揭示新型查耳酮衍生物MW-9抑制MAPK通路治疗溃疡性结肠炎的效应机制,为查尔酮衍生物MW-9治疗溃疡性结肠炎提供科学依据。方法建立细菌脂多糖诱导的小鼠巨噬细胞RAW264.7炎症模型,MTT法检测MW-9对细胞活度的影响;Griess法检测细胞NO含量;ELISA法检测细胞培养上清及血清中炎症因子IL-6、IL-1β和TNF-α的水平。构建葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠模型,记录体质量;HE染色检测结肠病理损伤程度;Western blot测定结肠组织MAPK通路关键蛋白p38、ERK1/2、JNK的磷酸化水平。结果MW-9明显抑制RAW264.7细胞NO的产生,其IC50为20.47 mg·L^(-1),抑制炎症因子IL-6、IL-1β和TNF-α的高表达(P<0.05)。MW9在低于6 mg·L^(-1)的浓度下无明显细胞毒性。MW-9治疗后,小鼠体质量下降减缓,可降低血清中促炎因子IL-6、IL-1β和TNF-α水平;与模型组比较,MW-9明显降低MAPK通路中磷酸化p38和ERK1/2蛋白的表达水平。结论新型查耳酮衍生物MW-9在体内外具有明显的抗炎活性,其治疗溃疡性结肠炎作用机制可能与抑制MAPK信号通路有关。

Aim To investigate the therapeutic effect of the MW-9 on ulcerative colitis(UC)and reveal the underlying mechanism,so as to provide a scientific guidance for the MW-9 treatment of UC.Methods The model of lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cells was established.The effect of MW-9 on RAW264.7 cells viability was detected by MTT assay.The levels of nitric oxide(NO)in RAW264.7 macrophages were measured by Griess assay.Cell supernatants and serum levels of inflammatory cytokines containing IL-6,TNF-αand IL-1βwere determined by ELISA kits.Dextran sulfate sodium(DSS)-induced UC model in mice was established and body weight of mice in each group was measured.The histopathological damage degree of colonic tissue was assessed by HE staining.The protein expression of p-p38,p-ERK1/2 and p-JNK was detected by Western blot.Results MW-9 intervention significantly inhibited NO release in RAW264.7 macrophages with IC50 of 20.47 mg·L^(-1) and decreased the overproduction of inflammatory factors IL-6,IL-1βand TNF-α(P<0.05).MW-9 had no cytotoxicity at the concentrations below 6 mg·L^(-1).After MW-9 treatment,mouse body weight was gradually reduced,and the serum IL-6,IL-1βand TNF-αlevels were significantly down-regulated.Compared with the model group,MW-9 significantly decreased the expression of p-p38 and p-ERK1/2 protein.Conclusions MW-9 has significant anti-inflammatory activities both in vitro and in vivo,and its underlying mechanism for the treatment of UC may be associated with the inhibition of MAPK signaling pathway.