升麻素衍生物抑制LPS诱导的RAW264.7细胞炎症反应及改善大鼠佐剂性关节炎  

Inhibition of LPS-Induced Inflammatory Response in RAW264.7 Cells and Amelioration of Adjuvant Arthritis in Rats by Cimifugin Derivatives

作　　者：马嘉欣 赵岩[1] 韩梅[1] 洪傲天 韩佳宏[1] 蔡恩博[1] 杨利民[1] MA Jiaxin;ZHAO Yan;HAN Mei;HONG Aotian;HAN Jiahong;CAI Enbo;YANG Limin(College of Traditional Chinese Medicines,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学,中药材学院,长春130118

出　　处：《中国细胞生物学学报》2024年第7期1408-1418,共11页Chinese Journal of Cell Biology

基　　金：国家中药材产业技术体系岗位科学家项目(批准号:CARS-21);吉林省科技发展计划(批准号:20210101217JC)资助的课题。

摘　　要：该文旨在设计并合成一系列升麻素衍生物,并筛选出活性较好的衍生物,并进一步对其抗类风湿性关节炎的作用机制进行初步研究。采用药物拼合原理合成升麻素有机酸酯衍生物和升麻素三氮唑衍生物,对其结构进行鉴定;筛选升麻素衍生物对脂多糖(LPS)诱导的RAW 264.7细胞的抗炎作用;并构建完全弗氏佐剂(CFA)诱导的大鼠佐剂性关节炎模型,测定大鼠体质量、踝关节肿胀度、关节炎指数、脏器指数、炎症因子和相关蛋白的表达水平,采用苏木素–伊红(HE)染色观察踝关节滑膜组织病理变化,最后将化合物与核心靶点进行分子对接验证。结果显示,共得到21个升麻素衍生物,经鉴定均为新化合物,在6.25μmol/L~50μmol/L药物浓度下,这些化合物对RAW264.7细胞存活率均在≥85%以上,有5个化合物NO抑制率的IC_(50)值优于升麻素和地塞米松(地塞米松IC_(50)为13.10μmol/L)。与空白组相比,模型组大鼠体质量极显著降低(P<0.001)、足趾肿胀度显著提高(P<0.01)、血清炎症因子水平显著提高(P<0.05)、脏器指数显著提高(P<0.05)。与模型组相比,化合物15高剂量组的大鼠体质量显著提高(P<0.01)、足趾肿胀度显著降低(P<0.01)、血清炎症因子水平显著降低(P<0.05)、脏器指数显著降低(P<0.01),其中化合物15高剂量组疗效较好,且优于甲氨蝶呤。与模型组相比,化合物15高剂量组关节滑膜病坏、炎性细胞浸润经给药后得到明显改善,此外该组大鼠踝关节滑膜组织中p-p38/p38表达水平显著降低(P<0.01)、p-ERK/ERK表达水平极显著降低(P<0.001)和Cleaved Caspase-3表达水平显著降低(P<0.01),化合物15与关键靶点具有较好的亲和力。该研究得出,化合物15可治疗由完全弗氏佐剂(CFA)诱导的大鼠佐剂性关节炎,其作用机制可能与调控MAPK信号通路有关。The purpose of this paper was to explore the design and synthesis of a series of cimifugin de-rivatives,and to screen out the derivatives with better activity,so as to further study the mechanism of action of their anti-rheumatoid arthritis.The anti-inflammatory effects of cimifugin derivatives on LPS(lipopolysaccharide)-induced RAW 264.7 cells were screened,and CFA(complete Frances adjuvant)-induced rat adjuvant arthritis model was constructed,and the body weight,ankle swelling,arthritis index,organ index and related inflammatory factors were measured,and the compounds were molecularly docked with the core targets.The pathological changes of ankle synovium were observed by HE(hematoxylin-eosin)staining.The results showed that a total of 21 cimifugin deriva-tives were obtained,all of which were identified as new compounds,and at the concentration of 6.25μmol/L~50μmol/L,these compounds more than 85%on the viability of RAW 264.7 cells,and the IC_(50) value of NO inhibition rate of 5 compounds was better than that of cimifugin and dexamethasone(Dex IC_(50) was 13.10μmol/L).Compared with the blank group,the rats in the model group showed highly significant decrease in body weight(P<0.001),significant increase in toe swelling(P<0.01),significant increase in serum inflammatory factor levels(P<0.05)and significant increase in organ indices(P<0.05).Compared with the model group,the body weight of rats in the high dose group of compound 15 was significantly increased(P<0.01);toe swelling was significantly decreased(P<0.01);serum in-flammatory factor level was significantly decreased(P<0.05);and organ index was significantly decreased(P<0.01)among which the high-dose compound 15 group had better efficacy and was better than methotrexate.Compared with the model group,compound 15 high dose group joint synovial disease and inflammatory cell infiltration signifi-cantly improved.In addition,the expression levels of p-p38/p38 were significantly reduced(P<0.01),p-ERK/ERK were highly significantly reduced(P<0.001)and Cleaved Ca

关 键 词：升麻素 衍生物 类风湿性关节炎 结构修饰 MAPK 

分 类 号：R285.5[医药卫生—中药学]