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贺守第, 孙晨霞, 廖威权, 廖康汉, 朱辉军, 黄胜光, 张剑勇, 谭宁.冬凌草及冬凌草甲素对类风湿关节炎血瘀证小鼠炎症模型的影响[J].湖南中医药大学学报,2024,44(11):1984-1990[点击复制] |
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| 冬凌草及冬凌草甲素对类风湿关节炎血瘀证小鼠炎症模型的影响 |
| 贺守第,孙晨霞,廖威权,廖康汉,朱辉军,黄胜光,张剑勇,谭宁 |
| (华中科技大学协和深圳医院中医风湿科, 广东 深圳 518000;广州中医药大学第四临床医院风湿科, 广东 深圳 518000) |
| 摘要: |
| 目的 研究中药冬凌草（Rabdosia Rubescens, RR）及其主要活性成分冬凌草甲素（Oridonin, ORI）对类风湿关节炎（rheumatoid arthritis, RA）血瘀证小鼠炎症模型的炎症影响，为RA血瘀证提供新的治疗药物。方法 将C57BL/6雌性小鼠随机分为对照组（Control组）、模型组（Model组）、低浓度RR组（L-RR组）、高浓度RR组（H-RR组）、低浓度ORI组（L-ORI组）、高浓度ORI组（H-ORI组），每组6只。除Control组外，其余5组小鼠双侧后肢足垫内注射1次10 μL乳化的完全弗氏佐剂，背部皮下注射0.1 mg/kg盐酸肾上腺素注射液2 h后，置于4 ℃冷水中游泳5 min，1次/d，连续7 d。L-RR组、H-RR组灌胃240、480 mg/（kg·d）的RR溶液，L-ORI组、H-ORI组腹腔注射20、40 mg/（kg·d）的ORI溶液，1次/d，连续14 d。每3天测量小鼠后足足掌的厚度；治疗14 d后，称量小鼠体质量，评估关节炎指数评分，ELISA法检测足趾血管内皮生长因子A（vascular endothelial growth factor A, VEGFA）、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素（interleukin, IL）-1β和IL-6浓度，HE染色、番红固绿染色检测踝关节组织破坏程度。结果 各组间体质量差异均无统计学意义（P＞0.05）。与Contorl组比较，Model组关节肿胀、关节炎症细胞浸润、滑膜增生、软骨被破坏，后足足掌厚度、关节炎指数评分及TNF-α、IL-6、IL-1β、VEGFA浓度均增加（P＜0.05）。与Model组比较，L-RR组、H-RR组、L-ORI组、H-ORI组关节肿胀症状减轻；H-RR组、L-ORI组、H-ORI组第22天后足足掌厚度降低（P＜0.05）；H-RR组、H-ORI组的关节炎指数评分降低（P＜0.05）；H-RR组、L-ORI组、H-ORI组TNF-α、IL-6、IL-1β浓度降低（P＜0.05）；H-RR组、L-ORI组、H-ORI组VEGFA浓度降低（P＜0.05）；L-RR组、H-RR组、L-ORI组、H-ORI组的踝关节组织中炎症细胞浸润、滑膜增生及软骨破坏均减少。结论 RR、ORI可改善血瘀证RA小鼠模型关节肿胀、后足足掌厚度，降低关节炎指数评分及炎症因子TNF-α、IL-6、IL-1β和VEGFA的浓度，并能减少踝关节组织中炎症细胞浸润、滑膜增生及软骨破坏，说明RR及其主要活性成分ORI具有治疗RA血瘀证炎症模型的作用，VEGFA可能作为RA血瘀证疗效评估的指标。 |
| 关键词: 冬凌草 冬凌草甲素 类风湿关节炎 血瘀证 炎症 |
| DOI：10.3969/j.issn.1674－070X.2024.11.007 |
| 投稿时间：2024-04-22 |
| 基金项目:广东省中医药管理局项目（20241255）;深圳市科技创新计划面上项目（JCYJ20230807115913027）;深圳市医疗卫生三名工程项目（SZZYSM202311003）;深圳市南山区卫生健康系统科技重大项目（NSZD2024013）;深圳市南山区科技项目（NS2021106，NS2021065）。 |
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| Effects of Rabdosia Rubescens and oridonin on the inflammatory model of rheumatoid arthritis with blood stasis pattern in mice |
| HE Shoudi, SUN Chenxia, LIAO Weiquan, LIAO Kanghan, ZHU Huijun, HUANG Shengguang, ZHANG Jianyong, TAN Ning |
| (Rheumatology Department of Chinese Medicine, Shenzhen Hospital Collaborating with Huazhong University of Science and Technology, Shenzhen, Guangdong 518000, China;Rheumatology Department, The Fourth Clinical Hospital of Guangzhou University of Chinese Medicine, Shenzhen, Guangdong 518000, China) |
| Abstract: |
| Objective To study the effects of Chinese medicine Rabdosia Rubescens (RR) and its main active ingredient oridonin (ORI) on inflammation in a mouse model of rheumatoid arthritis (RA) with blood stasis pattern, and provide new therapeutic drugs for the treatment of this disease. Methods The C57BL/6 female mice were randomized into Control group, Model group, low concentration RR (L-RR) group, high concentration RR (H-RR) group, low concentration ORI (L-ORI) group, and high concentration ORI (H-ORI) group，with six mice in each group. Except for Control group, the other five groups were injected with 10 μl of emulsified complete Freund's adjuvant into the foot pads of bilateral hind limbs once. Two hours after the subcutaneous injection of adrenaline hydrochloride 0.1 mg/kg on the back, they were placed in cold water at 4℃ for five minutes, once a day, for seven consecutive days. L-RR and H-RR groups were treated with RR solution at the doses of 240 mg/(kg·d) and 480 mg/(kg·d) respectively by gavage, while L-ORI and H-ORI groups were intraperitoneally injected with ORI solution at the doses of 20 mg/(kg·d) and 40 mg/(kg·d) respectively, once a day, for 14 consecutive days. The thickness of the hind paws of mice was examined every 3 days. After 14-day treatment, the body mass was measured and the arthritis index score was evaluated. Besides, ELISA was used to check the concentrations of vascular endothelial growth factor A (VEGFA), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in the toes of mice, while HE staining and safranin-fast green staining to assess the degree of the ankle joint tissue damage. Results There was no statistically significant difference in body mass among the six groups (P>0.05). Compared with Control group, Model group showed joint swelling, infiltration of inflammatory cells in the joints, synovial hyperplasia, and cartilage destruction with increases in the thickness of the hind paws, arthritis index score, as well as the concentrations of TNF-α, IL-6, IL-1β, and VEGFA (P<0.05). Compared with Model group, L-RR, H-RR, L-ORI, and H-ORI groups showed alleviated joint swelling; H-RR, L-ORI, and H-ORI groups had decreased hind paw thickness after 22 days (P<0.05); H-RR and H-ORI groups had reduced arthritis index scores (P<0.05); H-RR, L-ORI, and H-ORI groups had lower concentrations of TNF-α, IL-6, and IL-1β (P<0.05); H-RR, L-ORI, and H-ORI groups had lower VEGFA concentration (P<0.05); L-RR, H-RR, L-ORI, and H-ORI groups all had reduced inflammatory cell infiltration, synovial hyperplasia, and cartilage destruction in the ankle joint tissue. Conclusion For a mouse model of RA with blood stasis pattern, RR and ORI can alleviate the joint swelling, decrease the hind paw thickness, lower the arthritis index scores and the concentrations of inflammatory factors including TNF-α, IL-6, IL-1β, and VEGFA, and reduce the inflammatory cell infiltration, synovial hyperplasia, and cartilage destruction in the ankle joint tissue. This indicates that RR and its main active ingredient ORI have therapeutic effects on the inflammatory model of RA with blood stasis pattern, and VEGFA may serve as an indicator for evaluating the efficacy of treatments for RA with blood stasis pattern. |
| Key words: Rabdosia Rubescens oridonin rheumatoid arthritis blood stasis pattern inflammation |
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