基于NLRP3炎症小体探讨黄连厚朴汤治疗溃疡性结肠炎的作用机制  

Huanglian Houpo Decoction exerts therapeutic effects on ulcerative colitis by inhibiting NLRP3 inflammasome activation

作　　者：朱文瑞 赵天文 徐洪锋[1] Wen-Rui Zhu;Tian-Wen Zhao;Hong-Feng Xu(Department of Traditional Chinese Medicine Pharmacy,Shaoxing Hospital of Traditional Chinese Medicine,Shaoxing 312000,Zhejiang Province,China)

机构地区：[1]绍兴市中医院中药剂科,浙江省绍兴市312000

出　　处：《世界华人消化杂志》2023年第6期221-229,共9页World Chinese Journal of Digestology

基　　金：浙江省中医药科技计划项目,No.2023ZL728;浙江省药学会医院药学专项科研资助项目,No.2019ZYY09.

摘　　要：背景黄连厚朴汤(Huanglian Houpo Decoction,HHD)具有抗炎的药理作用,且能改善溃疡性结肠炎(ulcerative colitis,UC)所致肠道炎性损伤,但其潜在作用机制尚不清楚.目的以核苷酸结合寡聚化结构域样受体蛋白3(nucleotide binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体为切入点,探讨HHD对UC的作用机制.方法将雄性C57BL/6小鼠随机分为正常组、模型组、美沙拉嗪组、黄连厚朴汤低、中、高剂量组.除正常组外,采用3%葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导小鼠UC模型,造模的同时按分组给药.给药期间,每日进行小鼠的一般情况评价,计算疾病活动指数(disease activity index,DAI).给药结束后,取结肠和血清,测量结肠长度,评估结肠黏膜损伤指数(colon mucosa damage index,CMDI),以苏木素-伊红(hematoxylin-eosin,HE)染色观察结肠组织病理改变并进行病理学评分,酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)法检测血清中促炎因子白细胞介素1β(interleukin-1β,IL-1β)、白细胞介素6(interleukin-6,IL-6)及肿瘤坏死因子α(tumor necrosis factor,TNF-α)的水平,蛋白免疫印迹法(Western blot)法检测结肠组织NLRP3和半胱氨酸天冬氨酸蛋白水解酶1(cystein-asparate protease 1,Caspase-1)的蛋白表达水平.结果与正常组相比,模型组小鼠DAI评分显著升高(P<0.01),结肠长度明显缩短(P<0.01),CMDI评分显著升高(P<0.01),组织病理学评分显著降低(P<0.01),血清中IL-1β、IL-6及TNF-α水平显著增高(P<0.01),结肠组织中NLRP3与Caspase-1蛋白表达显著增多(P<0.01).与模型组相比,黄连厚朴汤各剂量组和美沙拉嗪组DAI评分显著改善(P<0.01),结肠长度明显恢复(P<0.05或P<0.01);黄连厚朴汤中、高剂量组及美沙拉嗪组CMDI评分明显下降(P<0.05或P<0.01),组织病理学评分明显降低(P<0.05或P<0.01);黄连厚朴汤各剂量组和美沙拉嗪组血清中促炎因子IL-1β、IL-6及TNF-α水平均明显降低(P<0.BACKGROUND Huanglian Houpo Decoction(HHD)has anti-inflam-matory effects,and can improve intestinal inflammatory injury caused by ulcerative colitis(UC),but the underlying mechanism remains unclear.AIM To investigate the mechanism of HHD in treating UC based on nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome activation.METHODS Male C57BL/6J mice were randomized into a control group,model group,mesalazine group,and low-,medium-,and high-dose HHD groups(HHD-H,HHD-M,and HHD-L,respectively).Except for the control group,all other groups had free access to 3%DSS solution to establish the UC model.Intragastric administration was started at the same time as modeling for 1 wk.During the experiment,the general mental state and disease activity index(DAI)score of mice were daily recorded.At the end of the treatment,colon and serum samples were collected,colon length was measured,and colon mucosa damage index(CMDI)score was calculated.The pathological changes in colon tissue were observed by hematoxylin-eosin(HE)staining and the histological damage score was evaluated.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumour necrosis factor-α(TNF-α)were measured by enzyme-linked immunosorbent assay(ELISA).The protein expression of NLRP3 and Caspase-1(cystein-asparate protease 1)in colon tissues was detected by Western blot.RESULTS Compared with the control group,the DAI score and CMDI score of the model group were significantly increased(P<0.01),while colon length was significantly shortened(P<0.01).The histological damage score was significantly reduced(P<0.01).The expression levels of IL-1β,IL-6,and TNF-αincreased significantly in serum(P<0.01).Furthermore,the modeling caused obvious pathological changes and up-regulated the expression of NLRP3 and Caspase-1 in the colon(P<0.01).Compared with the model group,the HHD groups and the mesalazine group showed significantly improved DAI scores(P<0.01).The colon length recovered significantly(P<0.05 or P<0.01).The CMDI score and

关 键 词：黄连厚朴汤 溃疡性结肠炎 NLRP3炎症小体 作用机制 

分 类 号：R285.5[医药卫生—中药学]