梓醇抑制骨关节炎发病机制的研究  

Study of mechanism for catalpol suppresses osteoarthritis in rats

作　　者：曾允富[1] 卞阳阳 王溶 彭磊[1] ZENG Yun-fu;BIAN Yang-yang;WANG Rong(Trauma Center, the First Affiliated Hospital of Hainan Medical College,Haikou, Hainan 570102, China)

机构地区：[1]海南医学院第一附属医院创伤中心

出　　处：《中华全科医学》2019年第10期1626-1630,1651,共6页Chinese Journal of General Practice

基　　金：国家自然科学基金项目(81460339)

摘　　要：目的探讨梓醇(CAT)对白介素-1β(IL-1β)诱导的软骨细胞炎症模型以及大鼠骨关节炎(OA)关节退变的机制研究。方法为探讨CAT在骨关节炎中的作用及其特定的作用机制,随机选取12只SD大鼠膝关节软骨细胞进行原代培养,将培养后的软骨细胞随机分成4组,即对照组(Control组)、IL-1β刺激组、IL-1β+CAT10μg/mL组、IL-1β+CAT50μg/mL组。动物实验中随机选取12只大鼠分为3组,Sham组、OA组及OA+CAT组,每组4只,OA组行右侧前交叉韧带横断和半月板切除术,实验组即OA+CAT组在切除完右侧前交叉韧带横断和半月板后,给予CAT[5mg/(kg·d)]腹腔注射,随机方法均采用抽签法。利用Western blotting方法、实时定量RT-PCR和免疫荧光染色检测软骨细胞的代谢、凋亡水平及相关信号通路。同时,应用番红-固绿染色和国际骨关节炎研究学会评分(OARSI)系统,对实验性大鼠软骨退变程度进行了评估。结果①CAT可抑制IL-1β诱导的软骨细胞凋亡;②CAT在软骨细胞中抑制IL-1β介导的分解代谢酶的释放,同时抑制IL-1β诱导的细胞外基质的降解;③CAT抑制IL-1β介导的NF-κB通路,同时减少IL-6、肿瘤坏死因子-α(TNF-α)的释放;④CAT在大鼠OA模型中可部分逆转软骨的退化。结论本实验结果提示,CAT能抑制NF-κB通路,减轻IL-1β诱导的大鼠软骨细胞的炎症反应和分解代谢水平,同时能抑制软骨细胞凋亡水平,所有的结果提示CAT具有治疗OA的作用。Objective To establish a IL-1β-induced rat chondrocyte inflammatory response model, and investigate the effects of catalpol(CAT) on cartilage degeneration of rats with osteoarthritis(OA). Methods To investigate the role of CAT in osteoarthritis and its specific mechanism, chondrocytes from 12 SD rats were randomly selected for primary culture. Chondrocytes were divided into four groups: control group, IL-1β group, IL-1β+CAT 10 μg/mL group and IL-1β+CAT 50μg/mL group. In animal experiments, 12 rats were randomly divided into three groups: Sham group, OA group and OA+CAT group, with 4 rats in each group. OA group underwent right anterior cruciate ligament transection and meniscectomy. The experimental group, namely OA+CAT group, was given intraperitoneal injection of CAT [5mg/(kg·d)] after resection of right anterior cruciate ligament transection and meniscus. Catabolic metabolism, apoptotic level and relative signaling pathway were measured by Western blotting, RT-PCR and immunofluorescence staining. We also assess the cartilage degeneration in an experimental rat model using Safranin O/fast green staining and Osteoarthritis Research Society International(OARSI) system. Results ①CAT prevented chondrocyte apoptotic level triggered by IL-1β;②CAT suppressed the release of catabolic enzymes, and inhibited the degradation of extracellular matrix induced by IL-1β;③CAT inhibited the nuclear factor Kappa B pathway, reduced the production of inflammatory cytokines(IL-6, TNF-α) in IL-1β-treated chondrocytes;④CAT partially reversed cartilage degeneration in the knee joint in animal model of OA. Conclusion CAT treatment can attenuate IL-1β-induced inflammatory response and catabolism in rat chondrocytes by inhibiting the NF-κB pathway, suggesting the therapeutic potential of CAT for the treatment of OA.

关 键 词：核因子-ΚB 炎症 梓醇 骨关节炎 凋亡 

分 类 号：R684.3[医药卫生—骨科学]