文献类型：期刊文献

中文题名：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

英文题名：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[1];高慧琴[1];吴国泰[1];李飒[1];王丽明[1,2]

第一作者：郝倩莹

机构：[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

第一机构：甘肃中医药大学

年份：2024

卷号：35

期号：5

起止页码：1034

中文期刊名：时珍国医国药

外文期刊名：Lishizhen Medicine and Materia Medica Research

收录：;北大核心:【北大核心2023】；CSCD:【CSCD_E2023_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：獐牙菜苦苷;灵仙新苷;组分配伍;类风湿关节炎;COX-2

外文关键词：Swertiamarin;Lucigenin;Component compatibility;Rheumatoid arthritis;COX-2

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节COX-2基因表达最为显著(P<0.01)。结论獐牙菜苦苷、灵仙新苷及其配伍对RA模型大鼠能起到良好的治疗作用,其中獐-灵配伍组的治疗效果明显优于各单体给药组;獐牙菜苦苷与灵仙新苷具有明显的协同增效作用,其作用机制可能是通过降低炎性细胞因子的分泌,抑制COX-2蛋白活性而发挥作用。
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 worse,and the AI score and serum levels of TNF-α,IL-1β,RF,CRP,anti-CCP Ab,and PGE2 were significantly higher than those in other groups(P<0.05 or P<0.01)in the left hind toe of rats;the ankle joint tissues and structures were severely damaged,and the expression of COX-2 proteins and genes were significantly up-regulated(P<0.01).Compared with the model group,the general state of rats in each dosing group showed different degrees of improvement;AI scores were significantly reduced(P<0.01),and serum levels of TNF-α,IL-1β,RF,CRP,anti-CCP,and PGE2 were significantly reduced(P<0.05 or P<0.01),among which the swertia-ling dosing group inhibited TNF-α,IL-1β,RF,CRP,anti-CCP and PGE2 were the most prominent;the histopathological status of ankle joints was improved to different degrees;IHC and WB test results showed that the swertia-spirit combination group reduced the protein expression level of COX-2 in ankle joints most significantly(P<0.01);and PCR results showed that swertia-spirit ascorbic acid group reduced the gene expression of COX-2 in ankle joints most significantly(P<0.01).Conclusion Swertia bittersweet,lingxian xinoside and their combination can play a good therapeutic effect on RA model rats,in which the therapeutic effect of the swertia-lingxian combination group was significantly better than that of the monomer administration group;swertia bittersweet and lingxian xinoside have obvious synergistic effect,and their mechanism of action may be to play a role in the reduction of the secretion of inflammatory cytokines,and the inhibition of the activity of the COX-2 protein.