目的 研究葡萄籽提取物（GSPE）与柳氮磺吡啶（SASP）联合用药治疗葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎的效果及作用机制。方法 将SPF级C57小鼠随机分为对照组、模型组、GSPE （250 mg/kg）组、SASP （250 mg/kg）组、联合用药（SASP 250 mg/kg+SASP 250 mg/kg）组，对照组给予正常饮用水，其他组均自由饮用3% DSS水溶液，连续饮用7 d，诱发小鼠溃疡性结肠炎模型；造模第1天同步开始ig给药，每天记录小鼠体质量、便血、便型等症状变化；7 d后断头取血，收集结肠和脾脏，记录结肠长度、脾脏质量变化情况。HE染色评估小鼠结肠黏膜组织病理变化，ELISA法检测血清和结肠组织中炎症因子肿瘤坏死因子（TNF-α）、白细胞介素（IL）-1β、IL-6和NO的表达变化以及丙二醛（MDA）、超氧化物歧化酶（SOD）细胞因子的含量变化，免疫组化分析结肠组织上皮细胞中NF-κB-p65、Nrf2、HO-1和Keap-1含量的变化。应用金氏Q值法分析联合用药的作用效果。结果 与模型组比较，各给药组的小鼠体质量下降幅度显著降低（P<0.01），小鼠腹泻、便血症状改善明显，其中联合用药组小鼠体质量较单独给药组下降更缓，小鼠腹泻、便血程度改善更明显（P<0.05）。各给药组小鼠血清及结肠组织中IL-1β、IL-6、TNF-α、NO、MDA含量较模型组显著降低，SOD含量则显著升高（P<0.01）；病理组织切片分析发现，各给药组小鼠结肠黏膜病理损伤较模型组显著减轻，其中联合用药组小鼠结肠黏膜病理损伤较单独给药组降低更为显著（P<0.05）。免疫组化分析结果也显示，各给药组小鼠的NF-κB-p65、Keap-1蛋白表达与模型组比较显著下调（P<0.01），Nrf2、HO-1蛋白表达显著上调（P<0.01）。联合给药组与单独给药组比较，NF-κB-p65、Keap-1蛋白表达显著降低（P<0.05），Nrf2、HO-1蛋白表达显著升高（P<0.05）。金氏Q值法评价两药合用后对IL-1β、IL-6、TNF-α、NO、MDA、NF-κB-p65、Keap-1的作用效果均大于1.15，说明GSPE与SASP具有协同作用。结论 GSPE与SASP联合用药治疗实验性溃疡性结肠炎效果优于单独给药，联合用药增强GSPE和SASP的抗氧化和抗炎作用，GSPE与SASP配伍治疗溃疡性结肠炎可进一步发挥协同增效的作用。

Objective To study the effect and mechanism of grape seed extract combined with sulfasalazine on DSS-induced ulcerative colitis (UC) in mice. Methods SPF grade C57 mice were randomly divided into five groups:control group, model group, grape seed extract group (GSPE, 250 mg/kg), sulfasalazine group (SASP, 250 mg/kg) and the combination medicine group[GSPE (250 mg/kg) + SASP (250 mg/kg)]. The control group was given normal drinking water, and the other groups were free to drink 3% DSS aqueous solution for 7 consecutive days to induce a model of UC in mice. The symptoms such as body weight, blood in the stool, and stool type were recorded every day. All mice were sacrificed after drug intervention for 7 consecutive days. Blood, colons and spleens were collected, respectively. The changes in colon length and spleen weight were recorded. HE staining was used to evaluate the pathological changes of mouse colonic mucosa. ELISA was used to detect the expression of inflammatory factors tumor necrosis factor:TNF-α, interleukin 1β (IL-1β), interleukin 6 (IL-6) and NO in serum and colon tissues, as well as the changes of MDA, SOD cytokines. Immunohistochemical analysis of the changes of content of NF-κB-p65, Nrf2, HO-1, Keap-1 were carried out in colon tissue epithelial cells. Effect of combined drug was evaluated by the Jin's Q-value method. Results Compared with model group, the body weight of mice in the GSPE group, SASP group and combination medicine group decreased slowly, and the symptoms of diarrhea and blood in the stool were significantly improved. The weight loss of the combination medicine group was slower than that of GSPE group and SASP group. Meanwhile, compared with the single administration groups, the diarrhea and bloody stools of the combination medicine group improved significantly. After administration, the contents of IL-1β, IL-6, TNF-α, NO, and MDA in the serum and colon tissues of mice were significantly lower than that in the model group, and the SOD content was significantly increased (P<0.01); pathological tissue analysis revealed that the pathological damage of the colonic mucosa of the mice in each administration group was significantly reduced, and the pathological damage of the colonic mucosa of the mice in the combination medicine extract was significantly lower than that of the other two groups (P<0.05). Immunohistochemical analysis showed that the expression of NF-κB and Keap-1 protein in each administration group was significantly reduced (P<0.01), and the expression of Nrf2 and HO-1 protein was significantly increased (P<0.05). Among them, the expression of NF-κB-p65 and Keap-1 protein in the combination medicine group was significantly lower than that in SASP group and GSPE group (P<0.05). The expression of Nrf2 and HO-1 protein in the combination medicine group were significantly higher than that in single administration groups (P<0.05). The Jin's Q-values of the effect of combined drug on IL-1β, IL-6, TNF-α, NO, MDA, NF-κB-p65, and Keap-1 were all greater than 1.15, which indicated that there is quite obviously synergy between GSPE and SASP. Conclusion Compared with single administration, the combined use of SASP and GSPE will enhance the antioxidant and anti-inflammatory effects of GSPE and SASP. The combined use of the two drugs has a synergistic effect on the treatment of UC.

R285.5

国家自然科学基金资助项目（81673693）；武警后勤学院创新团队基金项目（2019）；武警后勤学院博士启动金（WHB201710）；武警后勤学院基础研究项目（WHJ201905）