獐牙菜苦苷、灵仙新苷组分配伍对RA模型大鼠血清学指标及踝关节COX-2的影响  

Effects of swertiamarin bitter glycoside and lingxian new glycoside component pairing on serological indices and ankle COX-2 in RA model rats

作　　者：郝倩莹 高慧琴[1] 吴国泰[1] 李飒 王丽明 HAO Qian-ying;GAO Hui-qin;WU Guo-tai;LI Sa;WANG Li-ming(Gansu University of Chinese Medicine,Lanzhou Gansu,730101 China;Chinese People's Liberation Army 96604 Military Hospital,Lanzhou,Gansu 730031,China)

机构地区：[1]甘肃中医药大学,甘肃兰州730101 [2]中国人民解放军96604部队医院,甘肃兰州730031

出　　处：《时珍国医国药》2024年第5期1034-1040,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金地区基金项目(81960725)。

摘　　要：目的探讨獐牙菜苦苷、灵仙新苷组分配伍对类风湿关节炎(Rheumatoid arthritis,RA)模型大鼠血清学指标及踝关节COX-2的影响。方法选取SPF级SD大鼠5~6周龄36只,雌雄各半。除空白组外,均采用II型胶原诱导法复制(Collagen induced arthritis,CIA)大鼠模型。将造模成功的30只大鼠按随机数字表法分为模型组、阳性药(正清风痛宁)组、獐牙菜苦苷组、灵仙新苷组、獐牙菜苦苷配伍灵仙新苷组(以下简称“獐-灵配伍组”),每组6只。给药组分别给予相应药液灌胃,模型组和空白组给予等量生理盐水灌胃,连续灌胃10 d。实验中对各组大鼠进行一般状态观察记录、关节炎评分(AI),苏木素-伊红(HE)染色法观察大鼠踝关节组织病理变化;酶联免疫吸附测定法(ELISA)检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、类风湿因子(RF)、C反应蛋白(CRP)、抗环瓜氨酸肽抗体(anti-CCP Ab)及前列腺素E2(PGE2)含量;免疫组织化学法(Immunohistochemistry)及蛋白免疫印迹法(Western blot)检测大鼠踝关节环氧化酶-2(COX-2)蛋白表达;实时荧光定量聚合酶链式反应法(Real-time PCR)检测大鼠踝关节COX-2基因表达。结果与空白组比较,模型组大鼠一般状态较差,大鼠左后足趾AI评分及血清中TNF-α、IL-1β、RF、CRP、anti-CCP Ab、PGE2含量显著高于其他组(P<0.05或P<0.01);踝关节组织结构严重破坏,COX-2蛋白及基因表达显著上调(P<0.01)。与模型组比较,各给药组大鼠一般状态呈不同程度好转;AI评分明显降低(P<0.01),血清中TNF-α、IL-1β、RF、CRP、anti-CCP、PGE2含量明显降低(P<0.05或P<0.01),其中獐-灵配伍组抑制TNF-α、RF、CRP、anti-CCP、PGE2作用最为突出;獐牙菜苦苷在抑制IL-1β作用最突出。踝关节组织病理状态均有不同程度改善;IHC及WB检测结果显示獐-灵配伍组降低踝关节COX-2蛋白表达水平最为显著(P<0.01);PCR检测结果显示獐牙菜苦苷组降低踝关Objective Investigating the effects of swertiamarin and lingsenoside fractions on serological indices and ankle COX-2 in a rheumatoid arthritis(RA)model rat.Methods Thirty-six SPF-grade SD rats of 5-6 weeks of age,half male and half female,were selected.Except for the blank group,the rat model of Collagen induced arthritis(CIA)was replicated using the type II collagen induction method.The 30 rats successfully modeled were divided into the model group,the positive drug(Zhengqing Fengqin)group,the swertia bittersweet group,the lingxian xinsenoside group,and the swertia bittersweet with lingxian xinsenoside group(hereinafter referred to as the"swertia-lingxian group")according to the method of randomized numerical tables,and there were 6 rats in each group.The drug groups were given the corresponding liquid by gavage,and the model group and the blank group were given an equal amount of saline by gavage for 10 d.In the experiments,the general state of the rats in each group was observed and recorded,arthritis scores(AI)were recorded,and histopathological changes of the ankle joints of the rats were observed by hematoxylin-hematoxylin(HE)staining method;the serum tumor necrosis factor-α(TNF-α)and the tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA);the serum tumor nec rosis factor-α(TNF-α)and tumor necrosis factor-α(TNF-α)were measured by ELISA.Tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),rheumatoid factor(RF),C-reactive protein(CRP),anti-cyclic citrullinated peptide antibody(anti-CCP Ab)and prostaglandin E2(PGE2)in rat serum were detected by enzyme-linked immunosorbent assay(ELISA),and rat ankle joint cyclo-oxidation was detected by immunohistochemistry and protein immunoblot(Western blot).The protein expression of rat ankle cyclooxygenase-2(COX-2)was detected by immunohistochemistry and Western blotting;the gene expression of rat ankle COX-2 was detected by real-time PCR.Results Compared with the blank group,the general state of rats in the model group was w

关 键 词：獐牙菜苦苷 灵仙新苷 组分配伍 类风湿关节炎 COX-2 

分 类 号：R285.5[医药卫生—中药学]