摘要
目的:基于“太阴阴火”理论探讨升阳益胃汤对溃疡性结肠炎免疫信号Toll样受体(Toll-like Receptor,TLR)4/NOD样受体热蛋白结构域相关蛋白(NOD-like Receptor Thermal Protein Domain Associated Protein,NLRP)3通路的作用机制。方法:选用8周龄的C57BL/6小鼠随机分成6组,分别为正常组,模型组,西药组,升阳益胃汤低剂量组,升阳益胃汤中剂量组,升阳益胃汤高剂量组,观察小鼠的饮食、活动及身体毛发等状态;记录小鼠的体质量、粪便情况;评估小鼠结肠炎疾病活动指数;收集肛门到盲肠末端组织;采用苏木精-伊红技术检测小鼠结直肠病理变化;检测TLR4、髓性分化原发应答基因(88)[Myeloid Differentiation Primary Response Gene(88),MyD88]、NLRP3、半胱氨酸蛋白酶(Caspase,CASP)-1、Gasder min域(Gasder min domain,GSDMD)、人裂解的胱天蛋白酶-3(Cleaved CASP-3)蛋白表达水平。结果:①给药组各组小鼠饮食欲、体质量、精神状态、活动度、毛发、反应等情况都有好转,与模型组比较,西药组和各中药组的疾病活动指数评分下降较明显(P<0.05)。②模型组与正常组比较,小鼠的结直肠长度缩短,差异有统计学意义(P<0.05),各给药组与模型组比较小鼠结直肠长度有改善趋势,但差异无统计学意义。③升阳益胃汤中剂量组、升阳益胃汤高剂量组小鼠结肠上皮组织结构基本完整,腺体排列整齐,杯状细胞正常分布,肌层较模型组增厚,升阳益胃汤低剂量组小鼠结肠组织上皮结构基本完整,黏膜缺损减轻,腺体排列依旧紊乱,黏膜下层仍可见充血,炎性细胞浸润减少。④模型组与正常组比较,小鼠结直肠上皮组织TLR4,MyD88,核因子-κBp65,NLRP3,GSDMD、Cleaved CASP-3蛋白相对表达量增高(P<0.05);给药组与模型组比较,西药组、升阳益胃汤低剂量组,升阳益胃汤中剂量组,升阳益胃汤高剂量组小鼠结直肠上皮组织TLR4、MyD88、核因子-κB p65、NLRP3、GSDMD、Cleaved CASP-3蛋白相对表达量降低(P<0.05);与西药组相比,升阳益胃汤高剂量组MyD88、GSDMD、Cleaved CASP-3的蛋白相对表达量差异无统计学意义。结论:升阳益胃汤能够促进溃疡性结肠炎小鼠肠道损伤的修复。可以下调TLR4、MyD88、核因子-κB p65信号通路,抑制溃疡性结肠炎小鼠肠黏膜上皮的炎症及细胞焦亡反应。
Objective:To investigate the mechanism of the Shengyang Yiwei decoction(升阳益胃汤)on TLR4/NLRP3 pathway in ulcerative colitis based on the Yin(阴)fire in Taiyin(太阴)theory.Methods:Eight-week-old C57BL/6 mice were randomly divided into 6 groups:the normal group,the model group,the western medicine group,the low-dose Shengyang Yiwei decoction group,the middle-dose Shengyang Yiwei decoction group and the high-dose Shengyang Yiwei decoction group.The diet,activity and body hair of mice were observed,and the body weight and feces of mice were recorded.Disease activity index of colitis in mice was evaluated.The tissue from the anus to the end of the cecum is collected.The pathological changes of mouse rectum were detected by HE.The expression levels of TLR4,MyD88,NLRP3,CASP-1,GSDMD and Cleaved CASP-3 were detected.Results:①The appetite,body weight,mental state,activity,hair and reaction of the mice in the treatment groups were improved.Compared with the model group,the score of disease activity index in the western medicine group and the TCM medicine groups decreased significantly(P<0.05).Compared with the normal group,the colorectal length of mice in the model group was shorter,and the difference was statistically significant(P<0.05).②Compared with the model group,the colorectal length of mice in the treatment groups tended to improve,but the difference was not statistically significant.③The colonic epithelial tissue structure of mice in the middle dose group and high dose group of Shengyang Yiwei decoction are basically intact.The glands are arranged neatly,with normal distribution of goblet cells.The muscular layer was thicker than that in the model group.The structure of colonic epithelium in the low dose group of Shengyang Yiwei decoction is basically intact.Reduction of mucosal defect,and the arrangement of glands is still disordered.Congestion can still be seen in the submucosa,and the infiltration of inflammatory cells decreased.④Compared with the normal group,the relative expression of colorectal epithelial tissues TLR4,MyD88,nuclear factor-κBp65,NLRP3,GSDMD and Cleaved CASP-3 proteins increased(P<0.05).Compared with the model group,the relative expression of colorectal epithelial tissue TLR4,MyD88,NF-κBp65,NLRP3,GSDMD,and Cleaved CASP-3 proteins in the western medicine group,the Shengyang Yiwei decoction the low-dose group,Shengyang Yiwei decoction medium-dose group,and Shengyang Yiwei decoction high-dose group decreased(P<0.05).Compared with the western medicine group,there was no significant difference in the relative expression of MyD88,GSDMD and Cleaved CASP-3 in the high dose Shengyang Yiwei decoction group.Conclusion:The Shengyang Yiwei decoction can promote the repair of intestinal injury in mice with ulcerative colitis.It can down-regulate the signal pathways of TLR4,MyD88 and NF-κBp65,and inhibit the inflammation and cell death of intestinal mucosal epithelium in mice with ulcerative colitis.
出处
《中医临床研究》
2023年第32期81-86,共6页
Clinical Journal Of Chinese Medicine
基金
山西省卫生健康委科研课题,探讨升阳益胃汤对溃疡性结肠炎免疫信号TLR4/NLRP3通路介导的巨噬细胞M1型极化的抑制作用(2021086)
基于“太阴阴火”理论探讨升阳益胃汤对溃疡性结肠炎免疫信号TLR4/NLRP3通路的作用机制(2020057)。
