文献类型：期刊文献

中文题名：秦艽、威灵仙主要成分龙胆苦苷、木兰花碱对RA模型大鼠抗炎作用机制研究

英文题名：Anti-inflammatory Mechanism of Gentiopicroside and Magnoflorine of GENTIANAE MACROPHYLLAE RADIX and CLEMATIDIS RADIX ET RHIZOMA in Rats with Rheumatoid Arthritis

作者：葛珊[1];魏昀[1];吴晨[1];郑丽霞[1];李飒[1];郭从嘉[1];高慧琴[1]

第一作者：葛珊

机构：[1]甘肃中医药大学药学院,兰州730101

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2022

卷号：38

期号：4

起止页码：62

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金地区基金项目(编号:81960725);甘肃中医药大学研究生创新基金项目(编号:CX2020-37)。

语种：中文

中文关键词：龙胆苦苷;木兰花碱;类风湿关节炎;IκB激酶β;核因子-κB;环氧化酶-2

外文关键词：gentiopicroside;magnoflorine;rheumatoid arthritis;IKKβ;NF-κB;Cox-2

摘要：目的:探究秦艽、威灵仙中主要成分龙胆苦苷、木兰花碱及其配伍对类风湿关节炎(RA)模型大鼠的抗炎作用机制。方法:大鼠采用Ⅱ型胶原诱导法建立CIA大鼠模型,造模成功大鼠随机分为模型对照组、甲氨蝶呤1.05 mg/kg组、龙胆苦苷84.76 mg/kg组、木兰花碱0.5 mg/kg组、龙胆苦苷58.38 mg/kg-木兰花碱0.02 mg/kg组,另设正常对照组。各组分别灌胃给予相应药液或生理盐水,连续15 d。对大鼠进行一般状态观察记录、足跖肿胀度检测、关节炎指数(AI)评分;通过酶联免疫吸附技术(ELISA)检测大鼠血清中类风湿因子(RF)、C-反应蛋白(CRP)、肿瘤坏死因子-ɑ(TNF-ɑ)、白介素-1β(IL-1β)的含量;HE染色法观察各组大鼠关节组织病理学变化;实时荧光定量PCR技术(qRT-PCR)检测大鼠关节滑膜组织中IκB激酶β(Ikkβ)、核因子-κB(Nfkb)及环氧化酶-2(Cox2)mRNA的表达。结果:与正常对照组比较,模型对照组大鼠一般状态较差,足跖肿胀度及AI评分显著升高(P<0.01);血清中RF、CRP、TNF-ɑ、1L-1β的含量明显升高(P<0.05或P<0.01);滑膜组织中Ikkβ、Nfkb、Cox2 mRNA表达显著上调(P<0.01);关节组织结构破坏严重。与模型对照组比较,甲氨蝶呤1.05 mg/kg组、龙胆苦苷84.76 mg/kg组、木兰花碱0.5 mg/kg组和龙胆苦苷58.38 mg/kg-木兰花碱0.02 mg/kg组大鼠一般状态不同程度好转,大鼠足跖肿胀度及AI评分明显降低(P<0.05或P<0.01);血清中TNF-ɑ、IL-1β的含量显著降低(P<0.01),RF、CRP含量不同程度降低;滑膜组织中Ikkβ、Nfkb mRNA表达显著下调(P<0.01),不同程度下调Cox2 mRNA表达;关节组织病理变化不同程度改善。其中龙胆苦苷58.38 mg/kg-木兰花碱0.02 mg/kg组改善RA模型大鼠足跖肿胀情况、组织病理状态、抑制血清中CRP、TNF-ɑ、IL-1β分泌及对踝关节中Ikkb、Nfkb、Cox2 mRNA表达的下调作用最突出。结论:龙胆苦苷、木兰花碱及其配伍可对RA模型大鼠起到治疗作用,同时龙胆苦苷-木兰花碱的突出作用提示秦艽配伍威灵仙治疗RA具有一定优势。龙胆苦苷、木兰花碱及其配伍治疗RA的作用机制可能是通过降低炎性细胞因子分泌,抑制IKKβ/NF-κB/COX-2信号通路异常激活而发挥抗炎作用。
Objective: To investigate the anti-inflammatory mechanism of gentiopicroside and magnoflorine, the main components of GENTIANAE MACROPHYLLAE RADIX and CLEMATIDIS RADIX ET RHIZOMA, in rats with rheumatoid arthritis(RA). Methods: Rats were induced by type II collagen to establish the model of collagen-induced arthritis(CIA). The rats with successful modeling were randomly divided into a model group, a 1.05 mg/kg methotrexate group, a 84.76 mg/kg gentiopicroside group, a 0.5 mg/kg magnoflorine group, and a 58.38 mg/kg gentiopicroside+0.02 mg/kg magnoflorine group. The normal rats were divided into a control group. Each group was given the corresponding drug or normal saline for 15 days. The general state, paw swelling degree, and arthritis index(AI) score of rats were observed. The content of rheumatoid factor(RF), C-reactive protein(CRP), tumor necrosis factor-ɑ(TNF-ɑ), and interleukin-1β(IL-1β) in serum of rats in each group were detected by enzyme-linked immunosorbent assay(ELISA). Pathological changes in joints were observed by hematoxylin-eosin(HE) staining. Real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect the mRNA expression levels of IκB kinase β(IKKβ), nuclear factor kappa B(NF-κB), and cyclooxygenase 2(cox-2) in synovial tissues of rats in each group. Results: As compared with the control group, the general state of rats in the model group was poor, and the paw swelling degree and AI score were significantly increased(P<0.01). In the model group, the content of RF, CRP, TNF-ɑ, and IL-1β in serum were significantly increased(P<0.05 or P<0.01), the mRNA expressions of IKKβ, NF-κB, and cox-2 in synovial tissues were significantly up-regulated(P<0.01), and the joint structure was severely damaged. As compared with the model group, the general states of rats in the 1.05 mg/kg methotrexate group, the 84.76 mg/kg gentiopicroside group, the 0.5 mg/kg magnoflorine group, and the 58.38 mg/kg gentiopicroside+0.02 mg/kg magnoflorine group were improved to varying degrees, and the paw swelling degree and AI score were decreased(P<0.05 or P<0.01). In these groups, the content of TNF-ɑ and IL-1β in serum was decreased(P<0.01), and the content of RF and CRP was decreased to different degrees. The mRNA expressions of IKKβ and NF-κB in synovial tissues were down-regulated(P<0.01), and the mRNA expression of cox-2 was down-regulated to different degrees. The pathological changes in joints were improved to varying degrees. Among them, 58.38 mg/kg gentiopicroside+0.02 mg/kg magnoflorine group showed the optimal effect on the improvement of paw swelling degree and pathological changes in joints, decrease in the content of CRP, TNF-ɑ and IL-1β in the serum, and down-regulation of the mRNA expressions of IKKβ, NF-κB and Cox-2 in n synovial tissues. Conclusion: Gentiopicroside and magnoflorine can treat the rat model of RA. The evident effect of gentiopicroside-magnoflorine showed that GENTIANAE MACROPHYLLAE RADIX combined with CLEMATIDIS RADIX ET RHIZOMA had certain advantages in the treatment of RA, and the mechanism may be related to reducing the secretion of inflammatory cytokines, and inhibiting the excessive activation of IKKβ/NF-κB/cox-2 signaling pathway, thus playing an anti-inflammatory role.