香烟烟雾提取物对破骨细胞体积及整合素αvβ3基因表达的影响  

Effect of cigarette smoke extract on osteoclast size and gene expression of integrinαvβ3

作　　者：秦颖[1] 葛化冰[1] 刘琪 QIN Ying;GE Hua-bing;LIU Qi(Department of General Dentistry,Beijing Stomatology Hospital,Capital Medical University,Beijing 100006,China)

机构地区：[1]首都医科大学口腔医学院王府井院区口腔综合科,北京100006

出　　处：《北京口腔医学》2024年第3期184-189,共6页Beijing Journal of Stomatology

摘　　要：目的探讨香烟烟雾提取物(cigarette smoke extract,CSE)对破骨细胞体积及整合素αvβ3基因表达的影响。方法首先制作CSE溶液,实验采用7周龄C57BL/6J小鼠骨髓单个核细胞加入M-CSF和RANKL诱导生成破骨细胞,将浓度为5%的CSE加入实验组破骨细胞作用3 d,对照组不加入CSE。通过Trap染色识别对照组和实验组破骨细胞并于光镜下采图;通过甲苯胺蓝染色识别两组破骨细胞在骨片上形成的骨陷窝并于光镜下采图,均用NIH imageJ计算数量及面积。用实时荧光定量聚合酶链反应,以GAPDH作为内参,最后通过2-ΔΔCT计算,以检测Trap、CTR以及整合素αvβ3 mRNA表达水平。通过Western blot检测整合素αvβ3蛋白表达。结果破骨细胞表面积及细胞核数量的计量表明,实验组诱导形成的破骨细胞显著大于对照组(P<0.01)。实验组骨片上吸收陷窝数量及面积明显多于对照组(P<0.05),CSE处理组的TRAP mRNA水平为1.16±0.13,明显高于对照组(P<0.05),CTRmRNA水平为0.38±0.04,明显低于对照组(P<0.01)。CSE处理组的整合素αvβ3mRNA水平为1.75±0.05,显著高于对照组(P<0.01),蛋白质印迹分析结果也明显高于对照组(P<0.05)。结论CSE能显著增加破骨细胞的体积并上调整合素αvβ3的基因表达,并可能对骨吸收有促进作用。Objective To investigate the effect of cigarette smoke extract(CSE)on osteoclast size and gene expression of integrinαvβ3 in vitro.Methods 7-week-old C57BL/6J mouse bone marrow mononuclear cells were added with M-CSF and RANKL to induce osteoclasts.5%CSE was added to the experimental group for osteoclasts for 3 days,while no CSE was added in the control group.The osteoclasts in the control and experimental groups were identified using TRAP staining and images collected under light microscopy.The bone lacunae formed by osteoclasts on bone slices was identified using toluidine blue staining and images were taken under light microscopy.NIH imageJ was used to calculate the quantity and area.Real-time fluorescence quantitative polymerase chain reaction was used with GAPDH as the internal reference.The were detected by Trap,CTR,and integrinαvβ3 mRNA expression levels,and integrinαvβ3 protein expression was detected by 2-ΔΔCT calculation.The integrinαvβ3 protein expression was detected by Western blot.Results The induced osteoclasts in the experimental group were significantly larger than those in the control group(P<0.01).The number and area of absorption lacunae on the bone slices of the experimental group was significantly higher than that of the control group(P<0.05).The TRAP mRNA level in the CSE treatment group was 1.16±0.13,which was significantly higher than that of the control group(P<0.05).The CTR mRNA level was 0.38±0.04,which was significantly lower than that of the control group(P<0.01).Integrinsαvβ3 mRNA level in the CSE treatment group was 1.75±0.05,significantly higher than the control group(P<0.01),and the results of Western blot analysis were also significantly higher than the control group(P<0.05).Conclusions CSE can significantly increase the size of osteoclasts and upregulate the gene expression of integrinsαvβ3,which may be involved in the promotion of bone resorption.

关 键 词：香烟烟雾提取物 破骨细胞 骨吸收陷窝 整合素ΑVΒ3 细胞迁移 

分 类 号：R349.64[医药卫生—基础医学]