柚皮苷对破骨细胞凋亡的影响  

Effect of naringin on apoptosis of osteoclasts in vitro

作　　者：李风波[1] 孙晓雷[1] 马剑雄[1] 马信龙[1] LI Feng-bo;SUN Xiao-lei;MA Jian-xiong;MA Xin-long(Department of Orthopedics,Tianjin Hospital,Tianjin 300211,China)

机构地区：[1]天津市天津医院骨科,天津300211

出　　处：《中国矫形外科杂志》2021年第5期450-454,共5页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81871777,31600769)。

摘　　要：[目的]探讨不同浓度柚皮苷单体对破骨细胞的凋亡的影响及相关机制。[方法]采用100 ng/ml浓度RANKL诱导小鼠单核细胞RAW264.7细胞株获取破骨细胞,之后采用含有不同浓度柚皮苷培养基对破骨细胞进行干预3 d。行TRAP染色,扫描电镜观察骨薄片上骨吸收,流式细胞仪检测破骨细胞凋亡,荧光定量PCR检测破骨细胞凋亡基因BCL-2、BAX、 Caspase-3及MMP-9表达的影响。[结果]与空白对照组比较,各浓度组柚皮苷均可以有效的减少RANKL诱导破骨细胞的数量(P<0.05),减少薄骨片的吸收面积(P<0.05),且呈剂量依赖。流式细胞仪检测显示各浓度柚皮苷组可增加破骨细胞的凋亡率(P<0.05)。与空白对照相比,各柚皮苷组BCL-2和MMP-9的mRNA相对表达量降低(P<0.05),而BAX和Caspase-3的mRNA相对表达量升高(P<0.05),呈剂量依赖。[结论]柚皮苷可降低破骨细胞数量和骨吸收能力,这可能与下调BCL-2和MMP-9,上调BAX的caspase-3,增加破骨细胞凋亡有关。[Objective] To investigate the effect of naringin on osteoclast apoptosis in vitro, and explore the relative mechanisms.[Methods] RAW264.7 cells were induced to osteoclasts by 100 ng/ml RANKL for 5 days, and then the cells were intervened by naringin in different concentrations for another 3 days. Following assays were conducted, including TRAP staining for osteoclast, bone resorption on bone slice observed by scanning electron microscopy, osteoclast apoptosis assessed by flow cytometry, apoptosis-related gene mRNAs expression measured by real-time fluorescence quantitative PCR, such as BCL-2, BAX, Caspase-3 and MMP-9. [Results] Compared with the blank control group, the naringin in 3 groups with different concentrations significantly reduce the number of RANKL-induced osteoclasts by TRAP staining(P<0.05), and declined bone resorption area on bone slice(P<0.05) in dosage dependent manner. As result of flow cytometry, the naringin in 3 groups with different concentrations significantly increased the apoptosis rate of the osteoclasts compared with that of blank control group(P<0.05). In term of RT-qPCR assessment, the naringin in 3 groups with different concentrations significantly downgraded mRNA expressions of BCL-2 and MMP-9(P<0.05), whereas significantly upgraded mRNA expressions of BAX and Caspase-3 in dosage dependent manner in contrast with the blank control group(P<0.05). [Conclusion] The naringin considerably reduces the osteoclast number and bone resorption area on bone slice in dosage dependent manner this study in vitro, which might be related to down-regulate of BCL-2 and MMP-9, whereas up-regulate of BAX and caspase-3, and increase of osteoclast apoptosis.

关 键 词：柚皮苷 破骨细胞 细胞凋亡 骨质疏松 

分 类 号：R318[医药卫生—生物医学工程]