水甘草碱抑制破骨细胞活化缓解磨损颗粒诱导的炎性骨溶解  

Tabersonine alleviates wear particle-induced inflammatory osteolysis by inhibiting osteoclast activation

作　　者：张巍 俞磊 杨鹏[1] 耿德春[1] Zhang Wei;Yu Lei;Yang Peng;Geng Dechun(Department of Orthopedics,First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215006

出　　处：《中国组织工程研究》2024年第10期1519-1525,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(8227091833);项目负责人:耿德春。

摘　　要：背景:水甘草碱因其抗炎生物活性在心肌重塑、急性肾损伤和肺损伤等疾病中表现出良好的治疗作用。假体磨损颗粒常引发无菌性炎症,炎性因子的大量释放进一步导致了假体周围骨质的破坏和骨量丢失,然而目前尚无水甘草碱对假体周围骨溶解确切疗效的基础研究。目的:探讨水甘草碱对破骨细胞活化、炎症因子表达及磨损颗粒诱导炎性骨溶解的影响。方法:①细胞实验:将RAW264.7细胞分4组培养:对照组细胞加入完全培养基;破骨诱导组加入破骨诱导培养基(含50 ng/mL RANKL的完全培养基);水甘草碱低、高剂量组分别加入1,5μmol/L水甘草碱处理4 h后加入破骨诱导培养基。破骨诱导5 d后,分别进行抗酒石酸酸性磷酸酶染色、F-actin染色及RT-PCR检测。②动物实验:采用随机数字表法将20只C57BL/6J小鼠随机分为假手术组、骨溶解组、水甘草碱低及高剂量组,每组5只。骨溶解组、水甘草碱低及高剂量组通过将钛颗粒注射至颅骨表面的方法建立颅骨骨溶解模型,造模后第2天,水甘草碱低、高剂量组分别腹腔注射10,20 mg/kg水甘草碱,每隔1 d注射1次,直至造模2周后,收集小鼠动脉血血清进行炎性因子(白细胞介素1β、白细胞介素6、肿瘤坏死因子α)检测,收集颅骨进行micro-CT扫描和骨参数分析。结果与结论:①细胞实验:抗酒石酸酸性磷酸酶和F-actin染色显示,与破骨诱导组比较,水甘草碱低、高剂量组可抑制破骨细胞的活化及骨吸收,其中以高剂量组抑制更显著。RT-PCR检测结果显示,与对照组比较,破骨诱导组3种炎症因子的mRNA表达升高(P<0.01);与破骨诱导组比较,水甘草碱低及高剂量组3种炎症因子的mRNA表达降低(P<0.01),其中以高剂量组降低更明显。②动物实验:与假手术组比较,骨溶解组3种炎性因子水平升高(P<0.01);与骨溶解组比较,水甘草碱低及高剂量组3种炎性因子水平降低(P<0.05,P<0.01),�BACKGROUND:Tabersonine has shown good therapeutic effects in diseases such as myocardial remodeling,acute kidney injury and lung injury due to its antiinflammatory biological activity.Prosthetic wear particles often lead to aseptic inflammation,and the massive release of inflammatory factors further promotes periprosthetic bone destruction and bone loss;however,there are no basic studies on the efficacy of tabersonine on periprosthetic osteolysis.OBJECTIVE:To investigate the effects of tabersonine on osteoclast activation,expression of inflammatory factors and inflammatory osteolysis induced by wear particles.METHODS:(1)Cell experiment:RAW264.7 cells were divided into four groups for culture.A complete medium was added in the control group.Osteoclast induction medium(50 ng/mL RANKL+complete medium)was added to the osteoclast induction group.1 and 5μmol/L tabersonine was added for 4 hours,and then osteoclast induction medium was added to the low-and high-dose tabersonine groups,respectively.After 5 days of induction,tartrate-resistant acid phosphatase staining,F-actin staining and RT-PCR were performed.(2)Animal experiments:Twenty C57BL/6J mice were randomly divided into sham operation group,osteolysis group,low-dose tabersonine group and high-dose tabersonine group(n=5 per group).Skull osteolysis model of the skull was established by injecting titanium pellets on the skull surface in the osteolysis group,low-dose tabersonine group and high-dose tabersonine group.On day 2 after model establishment,mice in the low-dose and high-dose tabersonine groups received intraperitoneal injections of 10 and 20 mg/kg tabersonine every 2 days,respectively.2 weeks after surgery,mouse sera were collected for detecting inflammatory factors(interleukin 1β,interleukin 6,and tumor necrosis factorα),and cranial bones were collected for micro-CT scan and bone parameter analysis.RESULTS AND CONCLUSION:(1)Cellular experiments:Tartrate-resistant acid phosphatase staining and F-actin staining showed that compared with the osteoclast ind

关 键 词：水甘草碱 炎性因子 破骨细胞活化 磨损颗粒 假体周围骨溶解 

分 类 号：R459.9[医药卫生—治疗学] R336[医药卫生—临床医学] R684