摘要
背景 随着我国进入人口老龄化加速发展期,类风湿关节炎(RA)患病比例不断增加,给社会和家庭造成沉重负担.四妙汤中药物的有效成分确有改善 RA 的作用,但分析方剂的作用及机制缺乏相关研究.目的 观察四妙汤对胶原诱导法建立的类风湿关节炎大鼠的干预效果及其对维生素 D 系统及中性粒细胞胞外诱捕网(NETs)的影响.方法 2021 年 11 月—2023 年 1 月选取 SD 雌性大鼠 70 只,采用随机数字表法将大鼠分为空白组(Control 组)、模型组(Model 组)、甲氨蝶呤组(MTX 组)、维生素 D 组(VD 组)、四妙汤低剂量组(SMT-L 组)、四妙汤中剂量组(SMT-M 组)、四妙汤高剂量组(SMT-H 组),每组 10 只.通过胶原诱导法建立滑膜型关节炎大鼠模型,于造模 7 d 后开始灌胃治疗.观察药物干预前后大鼠踝部肿胀程度,测量关节炎指数(AI)评分.采用免疫组化法检测滑膜组织髓过氧化物酶(MPO)、肿瘤坏死因子 α(TNF-α)表达情况,检测大鼠血清中白介素 6(IL-6)、25-羟基维生素 D3[25(OH)D3]、NETs 水平,检测关节滑膜组织 24-羟基化酶(CYP24A1)、1-α 羟化酶(CYP27B1)、MPO、维生素 D 受体(VDR)、弹性蛋白酶(NE)表达水平.结果 药物干预第第 7 天 Control组、MTX 组、VD组、SMT-L 组、SMT-M 组、SMT-H 组踝部肿胀程度低于 Model 组,第 14 天 Model 组踝部肿胀程度高于 Control 组,SMT-M 组踝部肿胀程度高 于 Model 组,第 21 天 Model 组 踝部肿胀程度高 于 Control 组、MTX 组,低 于 VD 组、SMT-L 组、SMT-M 组、SMT-H 组,第 28 天 Control 组、MTX 组、VD 组、SMT-L 组、SMT-M 组、SMT-H 组踝部肿胀程度低于 Model 组(P<0.05).第14 天 SMT-H 组大鼠 AI 评分低于 Model 组,第 21 天 MTX 组、VD 组、SMT-L 组、SMT-H 组AI 评分低于 Model 组,第 28 天MTX 组、VD 组、SMT-L 组、SMT-M 组、SMT-H 组AI 评分低于 Model 组(P<0.05).7 组大鼠滑膜组织 MPO、TNF-α 表达水平比较,差异有统计学意义(P<0.05),其中 Model 组 MPO、TNF-α 高于 Control组,MTX 组、VD 组、SMT-H 组 MPO、TNF-α 低于 Model 组,SMT-M 组 TNF-α 低于 Model 组(P<0.05).Model 组 IL-6、NETs 高于 Control 组,25(OH)D3 低于 Control 组;MTX 组、VD 组、SMT-H 组 IL-6、NETs 低于 Model 组,25(OH)D3 高于 Model 组;SMT-M 组 NETs 低于 Model 组(P<0.05).Model 组 CYP24A1 高于Control 组、MTX 组、VD 组、SMT-M 组、SMT-H 组,CYP27B1、VDR 低于 Control 组、MTX 组、VD 组、SMT-M 组、SMT-H 组,MPO 高于 Control 组、MTX 组、VD 组、SMT-L 组、SMT-M 组、SMT-H 组,NE 高于Control 组、VD 组、SMT-M 组、SMT-H 组(P<0.05).结论 四妙汤可能通过调控维生素 D 系统抑制 NETs 的形成,进而治疗 RA.
Abstract
Background As China enters the period of accelerated development of population aging,the proportion of rheumatoid arthritis cases is increasing,causing a serious burden on society and families.The active ingredients in simiao decoction have improved effect,but there is a lack of relevant studies to analyze the action and mechanism of prescriptions.Objective To observe the intervention effect of simiao decoction on rats with rheumatoid arthritis established by collageninduced arthritis and its effect on vitamin D system and neutrophil extracellular traps(NETs).Methods From November 2021 to January 2023,70 SD female rats were selected.The rats were randomly divided into the blank group(control group),model group(model group),methotrexate group(MTX group),vitamin D group(VD group),Simiao decoction low-dose group(SMT-L group),Simiao decoction medium-dose group(SMT-M group)and Simiao decoction high-dose group(SMT-H group)by random number table method,with 10 rats in each group.The rat model of synovial arthritis was established by collagen induction method and the gavage treatment was started 7 d after modeling.The degree of ankle swelling was assessed,and the arthritis index(AI)score of rats in each group the levels of interleukin 6(IL-6),25-hydroxyvitamin D3[25(OH)D3],NETs in serum of rats and the expression levels of 24-hydroxylase(CYP24A1),1-α hydroxylase(CYP27B1),MPO,vitamin D receptor(VDR)and elastase(NE)in synovial tissue of joint were measured.Results The degree of ankle swelling in the control,MTX,VD,SMT-L,SMT-M and SMT-H groups was lower than that in the model group on day 7,the degree of ankle swelling in the model group was higher than the control group,the SMT-M group was higher than the model group on day 14,the degree of ankle swelling in the model group was higher than the control and MTX groups and lower than the VD,SMT-L,SMT-M and SMT-H groups on day 21,the degree of ankle swelling in the control,MTX,VD,SMT-L,SMT-M and SMT-H groups was higher than the model group,and the difference was statistically significant(P<0.05).The AI score of rats in the SMT-H group was lower than that in the model group on day 14,the AI score in the MTX,VD,SMT-L and SMT-H groups was lower than that in the model group on day 21,the score of rats in the MTX,VD,SMT-L,SMT-M,and SMT-H groups was lower than that in the model group on day 28,the difference was statistically significant(P<0.05).Immunohistochemical results showed significant differences in the expression levels of MPO and TNF-α in synovial tissue among the 7 groups(P<0.05),MPO and TNF-α in the model group were higher than those in the control group,while MPO and TNF-α in the MTX,VD and SMT-H groups were lower than those in the model group,TNF-α in the SMT-M group was lower than that in the model group,and the difference was statistically significant(P<0.05).IL-6 and NETs in the model group were higher than those in the control group,and 25(OH)D3 was lower than that in the control group.IL-6 and NETs in the MTX,VD and SMT-H groups were lower than those in the model group,and 25(OH)D3 was higher than that in the model group.NETs in the SMT-M group were lower than those in the model group,and the difference was statistically significant(P<0.05).CYP24A1 in the model group was higher than that in the control,MTX,VD,SMT-M and SMT-H groups,while CYP27B1 and VDR in the model group were lower than those in the control,MTX,VD,SMT-M and SMT-H groups,MPO was higher than that of the control,MTX,VD,SMT-L,SMT-M and SMT-H groups,and NE was higher than the control,VD,SMT-M and SMT-H groups,and the difference was statistically significant(P<0.05).Conclusion Simiao decoction may inhibit the formation of NETs by regulating the vitamin D system,thereby treating RA.
基金项目
国家自然科学基金资助项目(82260890)
教育厅滚动支持省属高校科研平台团队项目(黔教技2022023)
教育厅滚动支持省属高校科研平台团队项目(黔科合基础-ZK[2022]一般001)
维普
万方数据