逍遥散对雷公藤多苷致胃损伤大鼠胃黏膜IL-1β、TNF-α、NF-κB P65的调节机制

Regulation mechanism of Xiaoyao Powder on the expression of IL-1β,TNF-αand NF-κB P65 in rats’gastric mucosa injuried by triperygium glycosides

目的:研究逍遥散对雷公藤多苷致胶原诱导性关节炎(CIA)大鼠胃损伤的治疗效果及机制。方法:90只SD大鼠适应喂养1周,除正常组大鼠(n=19)外,其余大鼠均以弗氏完全佐剂和牛Ⅱ型胶原制备CIA大鼠模型。模型制备成功后,将成模大鼠随机分组:模型组、模型+雷公藤多苷组(雷公藤多苷组)、模型+雷公藤多苷+逍遥散组(逍遥散组)。正常组和模型组每天灌胃等体积0.9%氯化钠溶液,雷公藤多苷组和逍遥散组每天灌服雷公藤多苷混悬液(9.375 mg/kg)制备胃损伤模型大鼠,逍遥散组在灌服雷公藤多苷混悬液1 h前灌服逍遥散水煎液(19.270 g/kg),连续6周。观察各组大鼠体质量、摄食量及滑膜、踝关节、胃窦病理变化;酶联免疫吸附法测定大鼠血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量;免疫组化、实时荧光定量PCR法和蛋白质印迹法观察大鼠胃窦组织IL-1β、TNF-α、核因子-κB P65(NF-κB P65)基因及蛋白表达。结果:雷公藤多苷组大鼠在关节炎症状缓解同时,伴有明显胃损伤,胃窦腺上皮萎缩变短,毛细血管扩张、充血,黏膜固有层炎细胞浸润,壁细胞明显减少或消失,肠上皮化生明显,体质量增长及摄食量显著低于正常大鼠,血清中IL-1β、TNF-α含量显著升高(P<0.05),胃窦组织IL-1β、TNF-α、NF-κB P65蛋白与基因表达显著增加(P<0.05)。逍遥散干预后,大鼠体质量增长及摄食量增加,胃窦病理形态有不同程度改善,血清中IL-1β、TNF-α含量显著降低(P<0.05),胃窦组织IL-1β、TNF-α、NF-κB P65蛋白与基因表达显著降低(P<0.05)。结论:逍遥散可通过调节IL-1β、TNF-α、NF-κB P65异常表达发挥保护胃黏膜作用。

Objective:To explore the therapeutic effect and mechanism of Xiaoyao Powder on gastric injury induced by triperygium glycoside(TG)in rats with collagen induced arthritis(CIA).Methods:A total of 90 SD rats were fed for 1 week.Except the normal group(n=19),the other rats were fed with Freund's complete adjuvant and bovine typeⅡcollagen to establish the CIA rat model.After the model was prepared successfully,these model rats were randomly divided into model group,model+TG group(TG group),and model+TG+Xiaoyao Powder group(Xiaoyao Powder group).The normal group and model group were given the same volume of normal saline every day,the rats in the TG group and Xiaoyao Powder group were given TG suspension(9.375 mg/kg)every day to prepare the gastric injury model,and the rats in Xiaoyao Powder group were given Xiaoyao Powder Decoction(19.270 g/kg)1 hour before the administration of TG suspension.The experiment lasted for 6 weeks.To observe the body weight,food intake,and pathological changes of synovium,ankle joint and gastric antrum in each group;the contents of IL-1βand TNF-αin serum were observed by enzyme-linked immunosorbent assay;immunohistochemistry,quantitative Real-time PCR and Western Blot were used to observe the gene and protein expressions of IL-1β,TNF-αand NF-κB P65 in gastric antrum.Results:The symptoms of arthritis were relieved in TG group,but at the same time,there was obvious gastric injury,the glandular epithelium of the antrum was atrophied and shortened,the capillaries were dilated and congested,the lamina propria inflammatory cells of the mucosa were infiltrated,the parietal cells were significantly reduced or disappeared,and the intestinal metaplasia was obvious,the body weight and food intake of gastric injury model rats were significantly lower than those of normal rats,and the contents of IL-1βand TNF-αin serum were obviously increased(P<0.05),the protein and gene expressions of IL-1β,TNF-αand NF-κB P65 in gastric antrum were increased distinctly(P<0.05).After Xiaoyao Powder intervention,the body weight and food intake were increased,the pathological morphology of gastric antrum was improved in different degrees,and the contents of IL-1βand TNF-αin serum were apparently decreased(P<0.05),the protein and gene expressions of IL-1β,TNF-αand NF-κB P65 were clearly lessened(P<0.05).Conclusion:Xiaoyao Powder can protect the gastric mucosa by regulating the abnormal expression of IL-1β,TNF-αand NF-κB P65.