紫草素联合低强度脉冲式超声波调节ERK-JNK/NF-κB信号通路抑制关节炎的药理活性研究  

Research on pharmacological activity of shikonin combined with low⁃intensity pulsed ultrasound to modulate ERK⁃JNK/NF⁃κB signaling pathway to inhibit arthritis

作　　者：刘晓旭 刘金渝 周本源 范凯健 LIU Xiaoxu;LIU Jinyu;ZHOU Benyuan;FAN Kaijian(Department of Rehabilitation Science,the Second Affiliated Hospital of Soochow University,Suzhou 215000,China;De-partment of Pharmacy,Mental Health Center,Chongming District,Shanghai 202150,China;Department of Radiation Oncology,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]苏州大学附属第二医院康复科,江苏苏州215000 [2]上海市崇明区精神卫生中心药剂科,上海202150 [3]上海交通大学医学院附属仁济医院放疗科,上海200127

出　　处：《内科理论与实践》2024年第6期385-392,共8页Journal of Internal Medicine Concepts & Practice

基　　金：上海市崇明区“可持续发展科技创新行动计划”项目(CKY2024-63)。

摘　　要：目的:探讨紫草素(shikonin)对抑制滑膜细胞增殖及炎症反应的可能机制,并评估其联合低强度脉冲式超声波(low-intensity pulsed ultrasound shockwave,LI-PUS)治疗类风湿性关节炎(rheumatoid arthritis,RA)的可行性。方法:体外实验通过脂多糖(lipopolysaccharide,LPS)诱导小鼠滑膜细胞异常增殖。采用细胞计数试剂(cell counting kit-8,CCK-8)和酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测滑膜细胞的增殖情况和炎症因子以及抗氧化物的表达水平。细胞划痕实验检测滑膜细胞的迁移情况。免疫荧光检测滑膜细胞中核因子κB(nuclear factor kappa B,NF-κB)的表达情况。体内实验通过完全弗氏佐剂诱导小鼠RA模型。40只小鼠平均分为正常组、对照组、紫草素组、紫草素+LI-PUS联合治疗组。记录治疗第12天和24天的小鼠关节炎肿胀指数并检测血清中炎症因子水平。蛋白质印迹法(Western blotting)检测小鼠膝关节滑膜组织中c-Jun氨基端蛋白激酶(c-Jun Nterminal protein kinase,JNK)、胞外信号调节激酶(extracellular signal-regulated kinase,ERK)、NF-κB磷酸化水平。结果:紫草素抑制LPS诱导的滑膜细胞炎症细胞因子和氧化应激。在紫草素不同浓度的干预下,滑膜细胞的迁移均被显著抑制,NF-κB蛋白表达含量有所降低。动物实验结果显示,佐剂诱导的关节炎小鼠关节肿胀评分显著升高,紫草素干预后明显降低。紫草素+LI-PUS组治疗效果良好。紫草素抑制炎症细胞因子表达,白介素(interleukin,IL)-1β显著降低,紫草素+LI-PUS联合治疗抗炎作用极显著。蛋白质印迹法结果显示,对照组ERK、JNK、NF-κB磷酸化程度显著高于正常组,紫草素组、紫草素+LI-PUS联合治疗组均显著抑制,且紫草素+LI-PUS联合治疗组优于紫草素组。结论:紫草素可抑制LPS诱导的滑膜细胞异常增殖,且可通过ERK-JNK/NF-κB信号通路发挥抗关节炎活性,中药联合LI-PUS共同治疗优于�Objective To investigate the possible mechanism of shikonin in inhibiting synovial cell proliferation and inflammatory response and the effect of combining with low⁃intensity pulsed ultrasound(LI⁃PUS)in the treatment of rheumatoid arthritis.Methods Abnormal proliferation of mouse synoviocytes were induced by lipopolysaccharide(LPS)in vitro experiments.CCK⁃8 and ELISA were performed to detect the proliferation of synoviocytes and the expression levels of inflammatory factors and antioxidants.Cell scratch assay was used to detect the migration of synoviocytes.Immunofluorescence was used to detect the expression of nuclear factor(NF)⁃κB in synoviocytes.In vivo experiments,a mouse rheumatoid arthritis(RA)model was induced by complete Freund’s adjuvant.Forty mice were equally divided into 4 groups,which were normal group,model control group,shikonin perilla treatment group,and shikonin perilla+LI⁃PUS combination treatment group.The arthritic swelling indexes of the mice were recorded in day 12 and day 24 of treatment.At the end of the experiment,the serum levels of inflammatory factors were tested,and the expression levels of c⁃Jun N⁃terminal protein kinase(JNK),extracellular signal⁃regulated kinase(ERK),and NF⁃κB in the synovial tissues of the knee joints in mice were detected by Western blot.Results Shikonin inhibited the expression of inflammatory cytokines in LPS⁃induced synoviocytes.Shikonin intervention was able to significantly reverse LPS⁃induced oxidative stress in synoviocytes.The migration of synoviocytes were significantly inhibited under the intervention of different concentrations of shikonin.The level of NF⁃κB protein expression in synovial cells was reduced after the intervention of shikonin.The results of animal experiments showed that adjuvant⁃induced arthritis mice had significantly higher joint swelling scores,which could be significantly reduced after the intervention of shikonin.Shikonin+LI⁃PUS group showed good therapeutic effect on mice.Shikonin inhibited the ex

关 键 词：紫草素 低强度脉冲超声 关节炎 ERK-JNK/NF-κB 

分 类 号：R593.2[医药卫生—内科学]