溃疡宁抑制NOXs-ROS-NLRP3炎症小体信号通路影响溃疡性结肠炎大鼠炎症的实验研究  

Experimental study on ulcerative colitis in rats for kuiyangning inhibiting NOXs-ROS-NLRP3 inflammasome signal pathway

作　　者：杨镛 蔡枫[1] 李鹏[2] 丁旭枫 明溪[2] YANG Yong;CAI Feng;LI Peng;DING Xufeng;MING Xi(Affiliated Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属市中医医院,上海200071 [2]上海中医药大学,上海201203

出　　处：《国际检验医学杂志》2018年第18期2230-2233,共4页International Journal of Laboratory Medicine

基　　金：上海市卫生和计划生育委员会资助项目(2015HP1124)

摘　　要：目的研究溃疡宁抑制黏膜组织还原型辅酶Ⅱ氧化酶-活性氧簇-NOD样受体蛋白3(NOXs-ROSNLRP3)炎症小体信号通路影响溃疡性结肠炎大鼠炎症的机制。方法选取BALB/c大鼠45只,随机分为空白组、模型组、溃疡宁组,每组各15只,经右旋葡萄糖硫酸钠(DSS)诱导建立溃疡性结肠炎模型,空白组、模型组给予蒸馏水0.02mL/g,溃疡宁组给予溃疡宁10g/(kg·d),灌胃3周后取结肠黏膜组织,以荧光定量PCR法检测NLRP3、凋亡相关的斑点样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白水解酶-1(caspase-1)mRNA表达水平,采用鲁米诺化学发光法检测结肠黏膜组织活性氧自由基(ROS)水平,计算DPI抑制后的还原型辅酶Ⅱ(NADPH)消耗率分析NOXs活性,并对比3组组织病理学评分(HS)及血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平。结果 NLRP3、ASC、caspase-1的mRNA表达水平:空白组<溃疡宁组<模型组,3组上述指标两两比较,差异有统计学意义(P<0.05);ROS水平、NOXs活性:空白组<溃疡宁组<模型组,3组上述指标两两比较,差异均有统计学意义(P<0.05);溃疡宁组HS评分(4.21±0.59)分,血清IL-1β(70.45±8.19)pg/L,TNF-α(61.55±6.24)pg/L,低于模型组,(P<0.05),高于空白组(P<0.05),模型组和空白组上述指标比较,差异也有统计学意义(P<0.05)。结论溃疡宁可能通过抑制结肠黏膜组织NOXs-ROS-NLRP3炎症小体信号通路来降低IL-1β、TNF-α促炎因子表达,从而减轻溃疡性结肠炎大鼠以结肠黏膜损伤为主要表现的炎症。Objective To study the mechanism of Kuiyangning in the inhibition of mucosa tissues NADPH oxidases （NOXs）-reactive oxygen species （ROS）-NOD like receptor protein 3 （NLRP3） inflammasome signal pathway to affect the ulcerative colitis rats. Methods Totally 45 BALB/c rats were selected and randomly divided into the blank group,the mode group and the Kuiyanning group,and 15 rats in each group.The dextrose sodium sulfate （DSS） was induced to establish the ulcerative colitis mode,the blank group and the mode group were given distilled water with 0.02 mL/g,and the Kuiyanning group was given the Kuiyangning 10 g/（kg·d ）.After 3 weeks of stomach irrigation,the colonic mucosa tissues were collected,and the expression levels of NLRP3,apoptosis related spot like protein （ASC） and cysteine aspartic acid hydrolase -1 （caspase-1） mRNA were detected by fluorescent quantitative PCR,and Luminol chemiluminescence method was used to detect the activity oxygen free radicals （ROS） in colonic mucosa tissues,and the NADPH consumption rate was calculated after DPI inhibition and the activity of NOXs was analyzed,and the tissue histopathology score （HS） and serum interleukin-1β （IL-1β）,tumor necrosis factor α （TNF-α） were compared among the three groups. Results The mRNA expression levels of NLRP3,ASC and caspase-1 were：the blank group〈the Kuiyangning group〈the mode group,and there were significant differences between the any two groups among the three groups （ P 〈0.05）.The ROS level and NOXs activity were：the blank group〈the Kuiyangning group〈the mode group,and there were significant differences in the above indexes between the any two groups among the three groups （ P 〈0.05）.The HS score with （4.21±0.59） points and serum IL-1β with （70.45±8.19） pg/L and the TNF-α with （61.55±6.24） pg/L in the Kuiyanning group were lower than those in the mode group（ P 〈0.05）,but were higher than those in the blank group（ P 〈0.05）,and there were signifi

关 键 词：溃疡宁 NOXs-ROS-NLRP3炎症小体信号通路 溃疡性结肠炎 大鼠 

分 类 号：R-33[医药卫生] R285