刘玉欣 孙志娟 代荣琴 陈正立 梁闯 范洪

摘要 目的：探究雷公藤多苷對类风湿关节炎大鼠关节血清及组织Ras/Raf1表达的影响。方法：将60只SD大鼠随机分为对照组、模型组和干预组，每组20只。模型组与干预组建立大鼠风湿关节炎模型，干预组用雷公藤多苷干预，对照组和模型组给予等量生理盐水。以足趾肿胀度为模型标准和疗效判定指标，实验终点通过酶联免疫吸附试验法（ELISA）检测大鼠血清Ras、Raf1以及炎症介质肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）水平，实验终点通过Western Blotting检测大鼠滑膜组织Ras和Raf1蛋白表达量并对大鼠踝关节进行HE染色分析。结果：ELISA结果表明，与对照组比较，模型组血清Ras、Raf1、TNF-α、IL-6显著升高（均P<0.01），与模型组比较，干预组血清Ras、Raf1、TNF-α、IL-6显著降低（均P<0.01）;Western Blotting结果表明：与对照组大鼠比较，模型组大鼠滑膜组织Ras、Raf1显著升高（均P<0.01），与模型组比较，干预组大鼠滑膜组织Ras、Raf1显著降低（均P<0.05）。HE染色表明：与对照组比较，模型组病理总评分升高（P<0.01），与模型组比较，干预组病理总评分下降（P<0.05）。结论：雷公藤多苷可抑制大鼠血清炎症介质释放并降低外周血和关节组织中Ras、Raf1蛋白的表达，起到骨保护作用。

关键词 雷公藤多苷;类风湿关节炎;大鼠;滑膜组织;炎症;病理

Effects of Tripterygium Glycosides on Ras/Raf1 Expression in Rats with Rheumatoid Arthritis

LIU Yuxin1，SUN Zhijuan1，DAI Rongqin3，CHEN Zhengli2，LIANG Chuang1，FAN Hong1

（1 Department of Laboratory，Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine，Cangzhou 061000，China; 2 Department of Laboratory，the First Hospital of Hebei Medical University，Shijiazhuang，Hebei 050023，China; 3 Cangzhou Medical College，Cangzhou 061001，China）

Abstract Objective：To explore the effects of tripterygium glycosides on the expression of Ras/Raf1 in joint serum and tissues of rats with rheumatoid arthritis.Methods：A total of 60 SD rats were randomly divided into a control group（n=20），a model group（n=20） and an intervention group（n=20）.Rat rheumatoid arthritis model was established in model group and intervention group.The intervention group was intervened with tripterygium glycosides，and the control group and model group were given the same amount of normal saline.Take toe swelling as model standard and curative effect judgment index，at the end of the experiment，the rat serum levels of Ras，Raf1，TNF-α and IL-6 were detected by ELISA.At the end of the experiment，the protein expression of Ras and Raf1 in rat synovial tissue was detected by Western Blotting，and the rat ankle joint was analyzed by HE staining.Results：The results of ELISA showed that，compared with the control group，the levels of serum Ras，Raf1，TNF-α and IL-6 in the model group were increased significantly（P<0.01）.Compared with the model group，the levels of serum Ras，Raf1，TNF-α and IL-6 in the intervention group were significantly decreased（P<0.01）.The results of Western Blotting showed that，compared with the control group，Ras and Raf1 in rat synovial tissue in the model group were significantly increased（P<0.01），and compared with the model group，Ras and Raf1 in rat synovial tissue in the intervention group were significantly decreased（P<0.01）.He staining showed that，compared with the control group，the total score of pathology in the model group increased（P<0.01），and compared with the model group，the total score of pathology in the intervention group decreased（P<0.05）.Conclusion：Tripterygium glycosides can inhibit the release of serum inflammatory factors in rats and reduce the expression of Ras and Raf1 protein in peripheral blood and joint tissue of rats，which plays a role in bone protection.

Keywords Tripterygium glycosides; Rheumatoid arthritis; Rats; Synovial tissue; Inflammation; Pathology

中图分类号：R285.5文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.17.011

风湿性关节炎（Rheumatoid Arthritis，RA）是急性或慢性结缔组织炎症，在临床上较为常见。临床以滑膜组织过度增生、关节和肌肉游走性酸痛、红肿、疼痛为主要特征[1]。膜受体酪氨酸蛋白激酶信号传递途径Ras/Raf/MEK/ERK可以将细胞外信号转导至细胞内，调控细胞的增殖与分化、凋亡与炎症反应等重要生物学功能[2]。雷公藤多苷具有抗炎及抑制细胞免疫和体液免疫等作用，临床多用于风湿热瘀，毒邪阻滞所致的类风湿性关节炎[3]。本研究通过建立大鼠类风湿性关节炎模型，探究雷公藤多苷对类风湿性关节炎大鼠Ras/Raf1表达的影响。

1 材料与方法

1.1 材料

1.1.1 动物

8周龄雄性SPF级SD大鼠60只，体质量180～200 g，购于斯贝福生物技术有限公司，许可证号（SCXK 2019-0010）。饲养环境温度20～25 ℃，湿度55%，自由饮食、饮水。……