摘要
目的 探讨芹菜素对骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)/核因子κB受体活化因子(RANK)信号通路的调控作用及对创伤性骨折大鼠骨折愈合的影响。方法 采用闭合式股骨干骨折术构建创伤性骨折大鼠模型,将造模成功的大鼠随机分为模型组,芹菜素低、中、高剂量组和阳性对照组,每组10只,另取10只健康大鼠作为对照组。各组给予相应干预30 d。采用计算机断层扫描测定大鼠骨小梁密度和厚度,番红O-固绿染色观察大鼠软骨组织病理学变化,酶联免疫吸附试验检测大鼠血清中骨钙素(BGP)、碱性磷酸酶(ALP)、诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6水平,实时荧光定量PCR和蛋白质印迹法检测大鼠股骨组织OPG、RANKL、RANK mRNA和蛋白水平。结果 对照组大鼠软骨组织正常,结构完整;与对照组比较,模型组大鼠软骨组织出现严重损伤,骨小梁密度和厚度、血清BGP、ALP及股骨组织OPG mRNA和蛋白水平降低(P<0.05),血清iNOS、TNF-α、IL-6及股骨组织RANKL、RANK mRNA和蛋白水平升高(P<0.05);与模型组比较,芹菜素低、中、高剂量组大鼠软骨组织病理损伤改善,骨小梁密度和厚度、血清BGP、ALP及股骨组织OPG mRNA和蛋白水平依次升高(P<0.05),血清iNOS、TNF-α、IL-6及股骨组织RANKL、RANK mRNA和蛋白水平依次降低(P<0.05);芹菜素高剂量组与阳性对照组各项指标比较,差异无统计学意义(P>0.05)。结论 芹菜素可以减轻创伤性骨折大鼠炎症反应,促进骨折愈合,其机制可能与上调OPG表达,抑制RANKL/RANK信号通路有关。
Objective To investigate the effect of apigenin on osteoprotegerin(OPG)/receptor activator of nuclear factor-κb ligand(RANKL)/receptor activator of nuclear factor-κb(RANK)signaling pathway and fracture healing in rats with traumatic fracture.Methods The rat model of traumatic fracture was established by closed femoral shaft fracture.The rats were randomly divided into model group,low dose apigenin group,medium dose apigenin group,high dose apigenin group and positive control group,with 10 rats in each group.Another 10 healthy rats were used as control group.Each group was given corresponding intervention for 30 d.The density and thickness of trabecular bone were measured by computed tomography,and the pathological changes of cartilage were observed by safranin O-fast green staining.The serum levels of osteocalcin(BGP),alkaline phosphatase(ALP),inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein levels of OPG,RANKL,and RANK in rat femoral tissue were detected by real-time fluorescent quantitative PCR and Western blot.Results In the control group,the cartilage tissue was normal and the structure was intact.Compared with the control group,the cartilage tissue of rats in the model group was severely damaged,the density and thickness of bone trabecula,the levels of serum BGP and ALP,OPG mRNA and protein in femoral tissue were decreased(P<0.05),the levels of iNOS,TNF-α,IL-6 in serum,the RANKL,RANK mRNA and protein in femoral tissue were significantly increased(P<0.05).Compared with the model group,the pathological changes of cartilage tissue in the low,medium,and high dose apigenin groups were significantly improved,and the density and thickness of trabecular bone,serum BGP and ALP levels,and OPG mRNA and protein levels in femoral tissue were increased in sequence(P<0.05),the levels of iNOS,TNF-α,IL-6 in serum and RANKL,RANK mRNA and protein in femoral tissue were decreased in sequence(P<0.05).There was no significant difference between high dose apigenin group and positive control group(P>0.05).Conclusion Apigenin could reduce inflammatory response and promote fracture healing in rats with traumatic fracture.The mechanism may be related to up-regulation of OPG expression and inhibition of RANKL/RANK signaling pathway.
作者
陈月
苗存良
安志辉
杜蒙蒙
刘国峰
贾天阳
李盼
CHEN Yue;MIAO Cunliang;AN Zhihui;DU Mengmeng;LIU Guofeng;JIA Tianyang;LI Pan(Department of Hand and Foot Microsurgery,Handan Central Hospital,Handan,Hebei 056001,China)
出处
《国际检验医学杂志》
CAS
2023年第12期1446-1451,共6页
International Journal of Laboratory Medicine
基金
河北省医学科学研究课题计划项目(20201574)。
