【摘要】 目的 基于Toll样受体4(TLR4)-核苷酸结合寡聚化结构域样受体(NOD)信号通路探究溃结灵对溃疡性结肠炎(UC)大鼠肠黏膜修复和肠道自噬的影响。方法 将60只大鼠分为正常组、UC组(模型组)和溃结灵低、中、高剂量组，每组12只，除正常组外其余4组给予2,4,6-三硝基苯磺酸和乙醇溶液灌肠制备UC模型。正常组和UC组灌胃等量生理盐水，溃结灵低、中、高剂量组灌胃1.85、3.70、7.40 g/kg溃结灵，每日1次，连续12周。分析各组大鼠病变活动指数(DAI)评分、结肠长度、结肠黏膜损伤指数(CMDI)评分；检测血清D-乳酸(D-LAC)、二胺氧化酶(DAO)、血清和结肠组织肿瘤坏死因子-α(TNF-α);检测尿乳果糖(L)、甘露醇(M),用于计算尿L/M;采用Western blot法检测结肠组织自噬相关蛋白1(Beclin1)、自噬相关基因5(ATG5)、TLR4、TLR2、NOD样受体蛋白3(NLPR3)蛋白表达。结果 与正常组比较，治疗1、2、6和12周UC组DAI评分升高；与UC组比较，治疗1、2、6和12周溃结灵低、中、高剂量组DAI评分降低(P<0.05)。与正常组比较，UC组结肠长度明显缩短，CMDI评分、血清D-LAC和DAO、尿L/M、血清和结肠组织TNF-α及结肠组织TLR4、TLR2、NLPR3蛋白表达升高或增加，结肠组织Beclin1和ATG5表达减少；与UC组比较，溃结灵低、中、高剂量组结肠长度延长，结肠组织Beclin1、ATG5表达增加，CMDI评分、血清D-LAC和DAO、尿L/M、血清和结肠组织TNF-α及结肠组织TLR4、TLR2、NLPR3蛋白表达降低或减少(P<0.05)。溃结灵低、中、高剂量组上述指标比较差异有统计学意义(P<0.05),呈剂量效应关系。结论 溃结灵可通过调控TLR4-NOD信号通路改善UC大鼠肠道自噬，减轻结肠组织炎症反应，有助于肠黏膜修复，而高剂量组效果较优。更多还原

【Abstract】 Objective To investigate the effects of Kuijieling on intestinal mucosal repair and intestinal autophagy in ulcerative colitis(UC) rats based on Toll-like receptor 4(TLR4)-nucleotide-bound oligomeric domain-like receptor(NOD) signaling pathway. Methods A total of 60 rats were divided into normal group, UC group(model group), and low-dose, medium-dose and high-dose Kuijieling groups, with 12 rats in each group. Except the normal group, the other four groups were given 2, 4, 6-trinitrobenzene sulfonic acid and ethanol solution enema to prepare UC model. The normal group and UC group were given the same amount of normal saline, and the low-, medium-and high-dose Kuijieling groups were given the same amount of Kuijieling 1.85 g/kg, 3.70 g/kg and 7.40 g/kg, respectively, once a day for 12 consecutive weeks. Disease activity index(DAI) score, colon length, and colon mucosal damage index(CMDI) score in each group were analyzed. Serum D-lactic acid(D-LAC), diamine oxidase(DAO), serum and colon tissue tumor necrosis factor-α(TNF-α) were detected using detection kit. Lactulose(L) and mannitol(M) were detected, for calculating urine L/M. Western blot assay was used to detect the expressions of autophagy-related protein Beclin1, autophagy-related gene 5(ATG5), TLR4, TLR2 and NOD-like receptor protein 3(NLPR3) proteins in colon tissue. Results Compared with normal group, DAI scores in the UC group was higher at 1, 2, 6 and 12 weeks after treatment, and compared with UC group, DAI scores in low-, medium-and high-dose Kuijieling groups was significantly decreased at 1, 2, 6 and 12 weeks after treatment(P<0.05). Compared with normal group, colon length in UC group was significantly shortened, CMDI score, serum D-LAC and DAO, urinary L/M values, TNF-α in serum and colon tissue, and TLR4, TLR2, NLPR3 protein expressions in colon tissue were significantly increased, and Beclin1 and ATG5 in colon tissues were significantly decreased(P<0.05). Compared with UC group, colonic length was significantly prolonged in low-, medium-and high-dose Kuijieling groups, Beclin1 and ATG5 protein expression was significantly increased. CMDI score, serum D-LAC and DAO, urinary L/M values, TNF-α in serum and colon tissue, and TLR4, TLR2, NLPR3 protein expressions were significantly decreased(P<0.05). There were statistically significant differences in the above indexes among low-, medium-and high-dose Kuijieling groups in a dose-effect manner(P<0.05). Conclusion Kuijieling can improve intestinal autophagy in UC rats by regulating TLR4-NOD signaling pathway, alleviate colon tissue inflammation, and facilitate intestinal mucosal repair, and the effect of high-dose group was better.更多还原