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目的探讨复方蜥蜴散不同微粒组合剂干预大鼠溃疡性结肠炎(UC)模型TLRs/My D88信号通路中肠组织髓样分化因子88(MyD88)、肿瘤坏死因子受体相关因子6(TRAF6)蛋白表达、下游炎症因子肿瘤坏死因子-α(TNF-α)含量及其对UC的作用机制。方法雄性SD大鼠84只,随机分为6组,即正常对照组、模型组、柳氮磺吡啶片治疗组、蜥蜴散80目、100目、80+100目等量混合治疗组,分别编号A~F组。A组予0.9%Na Cl溶液灌肠处理,B~F组均采用三硝基苯磺酸(TNBS)灌肠诱导制备UC模型。造模成功后,治疗组(C~F组)分别给予对应药物灌胃治疗,A组及B组给予0.9%Na Cl溶液灌胃治疗,连续14 d,频次相同。14 d后麻醉全部大鼠,行腹主动脉取血,离心低温保存,取血后取结肠组织病变处包埋。采用HE染色观察大鼠结肠黏膜病理改变,采用免疫组化法检测My D88、TRAF6蛋白的原位表达,采用酶联免疫吸附试验(ELISA试验)检测血清TNF-α水平。结果与正常对照组比较,其他组My D88、TRAF6蛋白表达量及TNF-α水平升高,差异均有统计学意义(P<0.05);各治疗组与模型组比较,各项指标水平均下降,差异有统计学意义(P<0.05);复方蜥蜴散各治疗组与柳氮磺吡啶片治疗组比较,复方蜥蜴散80+100目组治疗效果最佳,差异有统计学意义(P<0.05),蜥蜴散80目、100目组与柳氮磺吡啶片治疗组比较,差异无统计学意义(P>0.05);复方蜥蜴散各治疗组间比较,80+100目等量混合组各项指标水平明显降低,差异有统计学意义(P<0.05)。结论复方蜥蜴散不同微粒组合剂能抑制TLRs/My D88信号通路下游信号传递分子My D88、TRAF6蛋白表达及降低下游炎症因子TNF-α水平,提示其治疗溃疡性结肠炎的主要作用机制在于阻断溃疡性结肠炎的炎症反应,调节免疫,恢复肠道屏障,维持肠道稳态,促进肠黏膜修复。
Objective To investigate the effects of different compounds of compound lizards on the expression of myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor related factor 6 (TRAF6) protein in the TLRs / My D88 signaling pathway of ulcerative colitis (UC) Expression and downstream inflammatory cytokine TNF-α content and its mechanism of action on UC. Methods Eighty-four male Sprague-Dawley rats were randomly divided into six groups: normal control group, model group, sulfasalazine treatment group, lizards scattered 80 mesh, 100 mesh, 80 + 100 mesh equal mixed treatment group, A ~ F group. Group A was treated with 0.9% NaCl solution for enema treatment, and rats in groups B to F were treated with trinitrobenzene sulfonic acid (TNBS) enema to induce UC model. After successful modeling, the treatment group (C ~ F group) were given the corresponding drug gavage treatment, A group and B group were given 0.9% NaCl solution for 14 days, the same frequency. After 14 days, all rats were anesthetized, blood was taken from the abdominal aorta, centrifuged and stored at low temperature. The pathological changes of colonic mucosa were observed by HE staining. The expression of My D88 and TRAF6 protein was detected by immunohistochemistry. The level of serum TNF-α was detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the normal control group, the levels of My D88 and TRAF6 protein and the levels of TNF-α increased in other groups (all P <0.05). Compared with the model group, the levels of each index decreased (P <0.05). Compared with sulfasalazine treatment group, compound lizards powder treatment group showed the best effect (P <0.05), and the difference was statistically significant ), Lizards scattered 80 mesh, 100 mesh group and sulfasalazine tablets treatment group, the difference was not statistically significant (P> 0.05); compound lizards powder each treatment group comparison, 80 +100 mesh equal mixture of items The index level was significantly lower, the difference was statistically significant (P <0.05). Conclusion Different compounds from Lizards can inhibit the expression of My D88 and TRAF6 protein downstream of TLRs / My D88 signaling pathway and decrease the level of TNF-α, which is a downstream factor of TLRs / My D88 signaling pathway, suggesting that the main mechanism of action of LPS in treating ulcerative colitis is blocking Inflammatory reaction of ulcerative colitis, regulation of immunity, restoration of intestinal barrier, maintenance of intestinal homeostasis, and promotion of intestinal mucosa repair.