文献类型：期刊文献

中文题名：秦艽、威灵仙组分配伍对类风湿关节炎模型大鼠血清炎症因子及踝关节NF-κB、VEGF表达的影响

英文题名：Effect of Component Compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma on Serum Inflammatory Factors and Expression of NF-κB and VEGF in Ankle Joint of Rats with Rheumatoid Arthritis

作者：郭从嘉[1];吴国泰[1];高慧琴[1];郑丽霞[1];李飒[1];王丽明[1];郝倩莹[1]

第一作者：郭从嘉

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：2023

卷号：29

期号：11

起止页码：53

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金地区基金项目(81960725)。

语种：中文

中文关键词：秦艽;威灵仙;组分配伍;类风湿关节炎;抗炎;核转录因子-κB;血管内皮生长因子

外文关键词：Gentianae Macrophyllae Radix;Clematidis Radix et Rhizoma;component compatibility;rheumatoid arthritis;anti-inflammation;nuclear factor-κB;vascular endothelial growth factor

摘要：目的:探讨秦艽、威灵仙组分配伍对类风湿关节炎(RA)模型大鼠的抗炎效果及作用机制。方法:选取5~6周龄SPF级SD大鼠72只,雌雄各半。除空白组外,均采用Ⅱ型胶原诱导法复制胶原诱导型关节炎大鼠模型(CIA)。将造模成功的64只大鼠按随机数字表法分为模型组、甲氨蝶呤组(0.375 mg·kg^(-1))、龙胆苦苷配伍木兰花碱组(150.4541 mg·kg^(-1)+5.0618 mg·kg^(-1))、龙胆苦苷配伍灵仙新苷组(150.4541 mg·kg^(-1)+16.4331 mg·kg^(-1))、獐牙菜苷配伍木兰花碱组(3.4558 mg·kg^(-1)+5.0618 mg·kg^(-1))、獐牙菜苷配伍灵仙新苷组(3.4558 mg·kg^(-1)+16.4331 mg·kg^(-1))、獐牙菜苦苷配伍木兰花碱组(9.3032 mg·kg^(-1)+5.0618 mg·kg^(-1))、獐牙菜苦苷配伍灵仙新苷组(9.3032 mg·kg^(-1)+16.4331 mg·kg^(-1)),每组8只,共9组。各组分别给予相应药液或生理盐水灌胃,连续给药15 d。实验中观察并记录各组大鼠一般状态;酶联免疫吸附测定法(ELISA)检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、类风湿因子(RF)、C反应蛋白(CRP)、前列腺素E2(PGE2)、抗环瓜氨酸肽抗体(anti-CCP Ab)含量;苏木素-伊红(HE)染色法观察各组大鼠踝关节组织病理学变化;免疫组化法(IHC)及蛋白免疫印迹法(Western blot)检测大鼠踝关节核转录因子-κB(NF-κB)、血管内皮生长因子(VEGF)蛋白含量;实时荧光定量聚合酶链式反应法(Real-time PCR)检测大鼠踝关节NF-κB、VEGF的mRNA表达。结果:与空白组比较,模型组大鼠一般状态较差,足跖肿胀显著;血清CRP、anti-CCP Ab、IL-1β含量显著升高(P<0.01);踝关节组织结构破坏严重;踝关节NF-κB、VEGF蛋白及mRNA表达显著上调(P<0.01)。与模型组比较,各给药组大鼠一般状态明显改善;血清TNF-α、IL-1β、RF、CRP、PGE2及anti-CCP Ab的含量均不同程度降低,其中龙胆苦苷配伍灵仙新苷降低TNF-α与IL-1β、龙胆苦苷配伍木兰花碱降低RF与CRP、獐牙菜苦苷配伍木兰花碱降低PGE2、獐牙菜苷配伍灵仙新苷降低anti-CCP Ab的效果优于其他组分配伍;踝关节组织病理变化均有不同程度改善;踝关节NF-κB、VEGF蛋白及mRNA表达明显下调(P<0.05,P<0.01),其中獐牙菜苦苷配伍灵仙新苷下调蛋白及mRNA表达作用最为显著。结论:秦艽、威灵仙组分配伍对RA模型大鼠能起到良好的治疗作用,二药组分配伍具有多途径、多靶点协同增效的作用,其中龙胆苦苷配伍木兰花碱、龙胆苦苷配伍灵仙新苷、獐牙菜苷配伍灵仙新苷、獐牙菜苦苷配伍木兰花碱及獐牙菜苦苷配伍灵仙新苷5种组分配伍模式具有潜在优势。其作用机制可能是通过降低炎性因子分泌及抑制NF-κB、VEGF蛋白和mRNA的异常表达而发挥作用。
Objective:To investigate the anti-inflammatory effect of the component compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma on the rat model of rheumatoid arthritis(RA)and the mechanism.Method:Seventy-two SPF-grade SD rats(male and female)aged 5 to 6 weeks were selected.Except the blank group,the rat model of collagen-induced arthritis(CIA)was replicated by the typeⅡcollagen induction method.The 64 rats after successfully modeling were randomly divided into model group,methotrexate group(0.375 mg·kg^(-1)),gentianoside with magnoflorine group(150.4541 mg·kg^(-1)+5.0618 mg·kg^(-1)),gentianoside with clematichinenoside AR group(150.4541 mg·kg^(-1)+16.4331 mg·kg^(-1)),sweroside with magnoflorine group(3.4558 mg·kg^(-1)+5.0618 mg·kg^(-1)),sweroside with clematichinenoside AR group(3.4558 mg·kg^(-1)+16.4331 mg·kg^(-1)),swertiamarin with magnoflorine group(9.3032 mg·kg^(-1)+5.0618 mg·kg^(-1)),and swertiamarin with clematichinenoside AR group(9.3032 mg·kg^(-1)+16.4331 mg·kg^(-1)),with 8 rats in each group.Each group was given the corresponding medicinal solution or normal saline by gavage for 15 d.During the experiment,the general status,of rats in each group were observed and recorded.Tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),rheumatoid factor(RF),C reactive protein(CRP),prostaglandin E2(PGE2),and anticyclic peptide containing citrulline antibody(anti-CCP Ab)in the serum of rats were measured by enzymelinked immunosorbent assay(ELISA).The histopathological changes in rat ankle joints were observed by hematoxylin-eosin(HE)staining.Immunohistochemistry(IHC)and Western blot were used to detect the protein expression of nuclear factor-κB(NF-κB)and vascular endothelial growth factor(VEGF)in rat ankle joints.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of NF-κB and VEGF in rat ankle joints.Result:Compared with those in the blank group,rats in the model group were in poor general conditions with significant foot-plantar swelling,and the content of CRP,antiCCP Ab,and IL-1βin the rat serum was significantly increased(P<0.01).In the model group,the tissue structure of the ankle joint was severely damaged,and the protein and mRNA expression of NF-κB and VEGF in the rat ankle joints were significantly up-regulated(P<0.01).As compared with the model group,the general status of rats in each administration group was significantly improved.The levels of serum TNF-α,IL-1β,RF,CRP,PGE2,and anti-CCP Ab were reduced to different degrees in these administration groups,among which the effects of the gentianoside with clematichinenoside AR group on down-regulating serum TNF-αand IL-1β,the gentianoside with magnoflorine group on down-regulating serum RF and CRP,the sweroside with magnoflorine group on down-regulating serum PGE2,and the swertiamarin with clematichinenoside AR group on lowering serum anti-CCP Ab were better than those of administration groups.The histopathological changes in the ankle joint were improved to different degrees.The protein and mRNA expression of NF-κB and VEGF in rat ankle joints in the administration groups was significantly down-regulated(P<0.05,P<0.01),and the swertiamarin paired with clematichinenoside AR group had the most significant effect.Conclusion:The component compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma exerts a good therapeutic effect on the rat model of RA,and the compatibility of components from the two medicines has a multi-channel,multi-target,and synergistic effect.The five component compatibility patterns,namely gentiobioside with magnoflorine,gentiobioside with clematichinenoside AR,sweroside with clematichinenoside AR,swertiamarin with magnoflorine,and swertiamarin with clematichinenoside AR,all have potential advantages.The mechanism may be related to the reduction of inflammatory factor secretion and the inhibition of abnormal protein and mRNA expression of NF-κB and VEGF.