Nepetoidin B对小鼠胶原诱导关节炎疗效及作用机制探讨  

Expation of the therapeutic effect and mechanism of Nepetoidin B on collagen-induced arthritis in mice

作　　者：孙耀宗 何韬 刘卓[1] 水芳 田瑞雪 唐葆青 张建辉 Sun Yaozong;He Tao;Liu Zhuo;Shui Fang;Tian Ruixue;Tang Baoqing;Zhang Jianhui(Department of Orthopaedics,the Second Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Orthopaedics,the 923rd Hospital of the People′s Liberation Army Liaison and Security Forces,Nanning 530021,China;Department of Nephrology,the Fifth Clinical College of Shanxi Medical University,Taiyuan 030012,China)

机构地区：[1]山西医科大学第二医院骨科,太原030001 [2]中国人民解放军联勤保障部队第九二三医院骨科,南宁530021 [3]山西医科大学第五临床医学院肾内科,太原030012

出　　处：《中华风湿病学杂志》2025年第3期213-218,I0002,共7页Chinese Journal of Rheumatology

基　　金：广西自治区卫生健康委自筹经费科研课题(Z-A20231059)。

摘　　要：目的探究Nepetoidin B对小鼠胶原诱导性关节炎(CIA)的疗效和可能的作用机制。方法雄性DBA/1小鼠采用随机数字表法分为4组,分别为阴性对照组、CIA组、甲氨蝶呤治疗(CIA+Meth)组、NepetoidinB治疗(CIA+NepB)组,每组6只。除对照组小鼠外,其余小鼠建立CIA模型。第21天开始按照分组进行治疗至第60天结束。治疗期间每3天评估关节炎症状;治疗结束后,苏木精-伊红(HE)染色评估关节病理变化;ELISA法和生化比色法检测血清IL-17,IL-6,丙二醛(MDA)和一氧化氮(NO)含量;蛋白印迹法检测关节组织中核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO1)通路。设置CIA小鼠用Nepetoidin B治疗后再用Nrf2抑制剂治疗组进行机制验证。符合正态分布方差齐的多组间数据采用单因素方差。分析两两比较采用,进一步t检验。结果与CIA组比较,CIA+MethB和CIA+NepB治疗后小鼠后肢肿胀程度明显减轻,关节炎评分显著下降[CIA+Meth组(5.2±1.3),CIA+NepB组(6.8±1.2)与CIA组(11.0±1.7)比较,t=6.69,P=0.004;t=5.00,P=0.009),关节组织病变程度减轻[CIA+Meth组(1.5±1.0),CIA+NepB组(2.2±0.8)与CIA组(4.0±0.9)比较,t=4.44,P<0.001;t=3.84,P=0.005);血清IL-17[CIA+Meth组(257±69)ng/ml,CIA+NepB组(279±103)ng/ml与CIA组(414±71)ng/ml比较,t=3.86,P=0.006;t=2.63,P=0.020],IL-6[CIA+Meth组(32±6)ng/ml,CIA+NepB组(44±5)ng/ml与CIA组(56±11)ng/ml比较,t=4.69,P<0.001;t=2.48,P=0.040),MDA[CIA+Meth组(22±4)μmol/L,CIA+NepB组(22±8)μmol/L与CIA组(34±11)μmol/L比较,t=2.77,P=0.038;t=2.29,P=0.049]和NO[CIA+Meth组(37±12)μmol/L,CIA+NepB组(37±11)μmol/L与CIA组(56±12)μmol/L比较,t=2.71,P=0.040;t=2.90,P=0.035]含量均显著降低。Nrf2(CIA+Meth组0.263±0.024,CIA+NepB组0.273±0.022与CIA组0.211±0.034比较,t=3.18,P=0.044;t=2.70,0.049)/HO1(CIA+Meth组0.524±0.021,CIA+NepB组0.501±0.014与CIA组0.453±0.033比较,t=3.95,P=0.006;t=3.41,P=0.032)通路显著升高。Nrf2抑制剂可以抵消Nepetoidin B治疗关节炎的作用(CIA+NepB组关节病理�ObjectiveTo investigate the therapeutic effect and potential mechanism of Nepetoidin B on rheumatoid arthritis(RA).MethodsDBA/1 mice were divided into four groups using the random number method,namely the control group,model group,methotrexate group,and Nepetoidin B group.The collagen-induced arthritis(CIA)model was prepared.Mice were treated from day 21th to day 60th.Arthritis symptoms were evaluated every three days during treatment.At the end of treatment,pathological changes of joint tissue were observed through HE staining.Serum IL-17,IL-6,MDA,and NO levels were measured using ELISA and biochemical colorimetric assays.The Nrf2/HO1 pathway in joint tissues was detected using western blot.A group of CIA mice was treated with Nepetoidin B,followed by an Nrf2 inhibitor to validate the mechanism.One-way analysis of variance was used to compare between multiple groups with homogeneity of variance,pairwise comparison using LSD-t test.ResultsThe study found that mice treated with methotrexate and Nepetoidin B exhibited a significant reduction in arthritis scores(CIA+Meth group 5.2±1.3,CIA+NepB group 6.8±1.2 vs.CIA group 11.0±1.7,t=6.69,P=0.004;t=5.00,P=0.009),and joint histopathology compared to the CIA mice(CIA+Meth group 1.5±1.0,CIA+NepB group 2.2±0.8 vs.CIA group 4.0±0.9,t=4.44,P<0.001;t=3.84,P=0.005).Additionally,there was a significant decrease in serum IL-17[CIA+Meth group(257±69)ng/ml,CIA+NepB group(279±103)ng/ml vs.CIA group(414±71)ng/ml,t=3.86,P=0.006;t=2.63,P=0.020],IL-6[CIA+Meth group(32±6)ng/ml,CIA+NepB group(44±5)ng/ml vs.CIA group(56±11)ng/ml,t=4.69,P<0.001;t=2.48,P=0.040),MDA[CIA+Meth group(22±4)μmol/L,CIA+NepB group(22±8)μmol/L vs.CIA group(34±11)μmol/L,t=2.77,P=0.038;t=2.29,P=0.049]and NO[CIA+Meth group(37±12)μmol/L,CIA+NepB group(37±11)μmol/L vs.CIA group(56±12)μmol/L,t=2.71,P=0.040;t=2.90,P=0.035]levels,and a significant elevation in the Nrf2(0.263±0.021,0.273±0.022 vs.0.221±0.034,t=3.18,P=0.044;t=2.70,P=0.049)/HO1(0.524±0.021,0.501±0.014 vs.0.453±0.033,t=3.95,P=0.006

关 键 词：关节炎 类风湿 氧化性应激 Nepetoidin B Nrf2/HO1通路 

分 类 号：R285.5[医药卫生—中药学]