摘要
目的研究金线莲苷对巨噬细胞极化、破骨细胞生成和骨溶解的作用。方法(1)细胞实验:牺牲C57BL/6J小鼠,提取骨髓腔BMMs细胞,使用10~70μmol/L金线莲苷处理BMMs细胞,CCK-8kit测定安全剂量。金线莲苷预处理BMMs细胞4 h,随后脂多糖(lipopolysaccharide,LPS)和干扰素-γ(interferon gamma,IFN-γ)诱导BMMs细胞24 h和72 h,免疫荧光测定极化标志物iNOS和ARG-1表达情况。BMMs细胞经预处理在RANKL诱导下进行破骨分化6 d,TRAP染色和牛骨板吸收实验检测金线莲苷对破骨细胞生成和骨吸收功能的影响。(2)动物实验:随机将20只C57BL/6J小鼠分为4组:假手术组、骨溶解组、金线莲苷低剂量组、金线莲苷高剂量组,于小鼠颅骨放置钛颗粒。术后1 d每隔3 d腹腔注射5 mg/kg、10 mg/kg金线莲苷,假手术组和骨溶解组注射等体积生理盐水。给药2周后,对颅骨进行micro-CT扫描及骨参数分析。结果(1)细胞实验:低于50μmol/L剂量的金线莲苷对BMMs无明显毒性作用,使用10μmol/L及50μmol/L作为高低剂量进行后续研究;金线莲苷可以提高巨噬细胞M2极化,缓解LPS诱导的M1极化;金线莲苷可抑制RANKL诱导的破骨细胞活化,并且这种抑制作用呈现剂量依赖性。(2)动物实验:钛颗粒导致小鼠颅骨溶解破坏,50μmol/L金线莲苷显著改善了颅骨骨微结构。结论金线莲苷可以促进改善巨噬细胞极化重编程,减少破骨细胞的活化,缓解钛颗粒诱导的颅骨破坏。
Objective To study the effects of Kinsenoside on macrophage polarization,osteoclastogenesis and osteolysis.Methods(1)Cellular experiments:C57BL/6J mice were euthanized,and BMMs were isolated from the bone marrow cavity.The BMMs were treated with varying concentrations(10-70μmol/L)of Anoectochilus roxburghii glycoside,with safe dosages determined using the CCK-8 assay.Cells were pretreated with Anoectochilus roxburghii glycoside for 4 hours,followed by stimulation with lipopolysaccharide(LPS)and interferon-gamma(IFN-γ)to induce BMMs polarization over 24 and 72 hours.Immunofluorescence staining was conducted to assess the expression of polarization markers,iNOS and ARG-1.For osteoclast differentiation studies,pretreated BMMs were stimulated with RANKL for 6 days,after which TRAP staining and bovine bone resorption assays were utilized to evaluate the influence of Anoectochilus roxburghii glycoside on osteoclastogenesis and bone resorptive activity.(2)Animal experiments:20 C57BL/6J mice were randomly assigned into four experimental groups:the sham group,osteolysis group,low-dose Anoectochilus Roxburghii Glycoside group,and high-dose Anoectochilus Roxburghii Glycoside group.Titanium particles were implanted onto the calvarial bone surface.Beginning one day post-surgery,intraperitoneal injections of Anoectochilus Roxburghii Glycoside at doses of 5 mg/kg or 10 mg/kg were administered every three days,while the sham and osteolysis groups received equivalent volumes of saline.After a two-week treatment period,micro-CT scanning and subsequent bone parameter analysis were performed on the calvarial samples to evaluate the osteoprotective effects of Anoectochilus Roxburghii Glycoside.Results(1)Cellular experiments:the dose of Kinsenoside below 50μmol/L had no significant toxic effect on BMMs,and 10μmol/L and 50μmol/L were used as the high and low doses for the follow-up studies.Kinsenoside could increase the M2 polarization of macrophages and alleviate the LPS-induced M1 polarization.Kinsenoside could inhibit the RANKL-induced activation of osteoclasts,and this inhibition showed a dose-dependent effect.(2)Animal experiments:Titanium particles led to the destruction of cranial osteolysis in mice,and 50μmol/L Kinsenoside significantly improved the bone microstructure of cranial bone.Conclusion Kinsenoside promotes improved macrophage polarization reprogramming,reduces osteoclast activation,and attenuates Titanium particle-induced cranial bone destruction.
作者
陶云霞
张巍
李文豪
杨鹏
TAO Yun-xia;ZHANG Wei;LI Wen-hao;YANG Peng(Department of Orthopedics,First Affiliated Hospital of Soochow University,Suzhou,Jiangsu,215006,China;Department of Orthopedics,Suzhou Hospital,Nanjing Medical University,Suzhou,Jiangsu,215006,China)
出处
《中国血液流变学杂志》
2025年第1期24-28,66,F0002,共7页
Chinese Journal of Hemorheology
基金
2022年苏州市“科教强卫”科研项目(SKY2022117)。
关键词
骨溶解
极化
破骨细胞生成
金线莲苷
osteolysis
polarization
osteoclastogenesis
Kinsenoside
