摘要
背景:破骨细胞过度活化是骨质疏松症发生、发展过程中的关键环节,探究破骨细胞功能、开发具有抑制破骨细胞分化减少骨重吸收的药物,是当前探索骨质疏松症临床治疗的重要手段。目的:基于MAPK/核因子κB信号通路,分析帕罗西汀影响破骨细胞分化的作用机制。方法:①体外实验:采用CCK-8实验检测不同浓度帕罗西汀对小鼠骨髓源性巨噬细胞增殖的影响。在巨噬细胞集落刺激因子和核因子κB受体活化因子配体的诱导下,使用不同浓度帕罗西汀干预破骨细胞分化,抗酒石酸酸性磷酸酶染色确定破骨细胞数目;RT-qPCR检测帕罗西汀对破骨细胞分化相关细胞因子mRNA表达的影响;Western Blot检测帕罗西汀对小鼠骨髓源性巨噬细胞中核因子κB、MAPK信号通路及破骨细胞相关蛋白表达的影响。②体内实验:将40只C57BL/6小鼠按随机数字表分为空白对照组、脂多糖组、帕罗西汀2 mg/kg,5 mg/kg组,每组10只。脂多糖组、帕罗西汀2 mg/kg,5 mg/kg组每2 d颅骨矢状缝皮下注射脂多糖(构建小鼠颅骨骨溶解模型),帕罗西汀两组每次皮下注射脂多糖1 d后注射对应剂量的帕罗西汀,14 d后,分离颅骨进行Micro-CT扫描分析。结果与结论:①体外实验:当帕罗西汀浓度≤5μmol/L时对小鼠骨髓源性巨噬细胞的增殖无影响,浓度大于10μmol/L时抑制细胞的增殖;当帕罗西汀浓度≥0.5μmol/L时,可明显抑制破骨细胞分化的数量及形态,抑制破骨细胞分化c-Fos、Nfatc1、Ctsk、Mmp9、Acp5、Atp6v0d2的mRNA表达水平;0.5μmol/L的帕罗西汀可通过抑制核因子κB、MAPK信号通路的激活,进而抑制破骨细胞相关蛋白c-Fos、Nfatc1、Ctsk的表达;②体内实验:Micro-CT扫描分析结果显示,与脂多糖组比较,帕罗西汀5 mg/kg组可以有效提高骨小梁数量、骨体积分数(P<0.05),降低骨表面积骨体积比、骨小梁分离度(P<0.05);③结果表明,帕罗西汀可以通过抑制破骨细胞
BACKGROUND:Excessive activation of osteoclasts is one of the key links in the occurrence and development of osteoporosis.Exploring the function of osteoclasts and developing drugs that can inhibit osteoclast differentiation and reduce bone resorption is an important means to explore the clinical treatment of osteoporosis.OBJECTIVE:To analyze the mechanism of paroxetine on osteoclast differentiation based on the mitogen-activated protein kinase/nuclear factor-κB signal pathway.METHODS:(1)In vitro experiment:Different concentrations of paroxetine were used to interfere with bone marrow-derived macrophages in mice,and then the effect of paroxetine on cell activity was determined by cell counting kit-8.Under the induction of macrophage colony stimulating factor and receptor activator of receptor activator of nuclear factor-κB ligand,different concentrations of paroxetine were used to interfere with osteoclast differentiation.The number of osteoclasts was determined by tartrate-resistant acid phosphatase staining.The effect of paroxetine on the mRNA expression of osteoclast differentiation related cytokines was detected by real-time fluorescence quantitative real-time PCR.The effect of paroxetine on nuclear factorκB and mitogenactivated protein kinase signal pathways in the cells was detected by western blot.(2)In vivo experiment:Forty male C57BL/6 mice were randomly divided into four groups(n=10 per group):blank control group,lipopolysaccharide group,paroxetine 2 mg/kg treatment group,and paroxetine 5 mg/kg treatment group.Lipopolysaccharide was injected subcutaneously into the sagittal suture of the skull every 2 days to construct the mouse skull osteolysis model in the lipopolysaccharide group,paroxetine 2 mg/kg and 5 mg/kg groups.The corresponding dose of paroxetine was injected 1 day after each lipopolysaccharide injection in the two paroxetine groups.After 14 days of treatment,mouse skulls were isolated for Micro-CT scanning.RESULTS AND CONCLUSION:(1)In vitro experiment:there was no significant effect on the
作者
莫耀民
刘槃
马瑞鑫
曾高峰
宗少晖
Mo Yaomin;Liu Pan;Ma Ruixin;Zeng Gaofeng;Zong Shaohui(Department of Spine Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530000,Guangxi Zhuang Autonomous Region,China;Collaborative Innovation Centre of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry,Nanning 530000,Guangxi Zhuang Autonomous Region,China;School of Public Health,Guangxi Medical University,Nanning 530000,Guangxi Zhuang Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第32期5184-5190,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(81860402),项目负责人:宗少晖
广西医学高层次骨干人才培养“139”计划(桂卫科教发[2020]15号),项目负责人:宗少晖。
