CAJ | 학술논문

目的：探讨Jagged1通过活化Notch通路对RAW 264畅7细胞向破骨分化及增殖的影响。方法：将RAW 264畅7细胞分三组培养：对照组（细胞培养基＋鼠RANKL 50 ng／L）、Jagged1组（细胞培养基＋鼠RANKL ＋Jagged1重组蛋白）、γ-分泌酶抑制剂（ DAPT）组（细胞培养基＋鼠RANKL ＋Jagged1重组蛋白＋DAPT），实时荧光定量PCR检测各组破骨细胞标志基因组织蛋白酶K（Cathepsin K，CK）、抗酒石酸性磷酸酶（Tartrate-resistant acid phosphatase， TRAP）、降钙素受体（Calcitionin receptor， CTR）及Notch靶基因HES-1、HEY-1 mRNA的表达，TRAP染色鉴定破骨细胞，扫描电镜检测破骨细胞溶骨功能。免疫荧光检测NICD的表达变化，CCK-8检测RAW 264畅7细胞增殖。结果：Jagged1组TRAP、CK、CTR 及HES-1、HEY-1 mRNA 的表达、TRAP＋细胞数较对照组及DAPT组明显升高（P＜0畅05），而DAPT组与对照组均无明显变化；细胞免疫荧光显示Jagged1组NICD除表达于细胞膜、细胞质外，细胞核也有较高表达，对照组及DAPT组细胞核中NICD无明显表达；细胞培养48 h，Jagged1组细增殖较对照组及DAPT组出现明显抑制。结论：Jagged1通过活化Notch通路促进RAW 264畅7向破骨细胞分化、抑制其增殖。
목적：탐토Jagged1통과활화Notch통로대RAW 264창7세포향파골분화급증식적영향。방법：장RAW 264창7세포분삼조배양：대조조（세포배양기＋서RANKL 50 ng／L）、Jagged1조（세포배양기＋서RANKL ＋Jagged1중조단백）、γ-분비매억제제（ DAPT）조（세포배양기＋서RANKL ＋Jagged1중조단백＋DAPT），실시형광정량PCR검측각조파골세포표지기인조직단백매K（Cathepsin K，CK）、항주석산성린산매（Tartrate-resistant acid phosphatase， TRAP）、강개소수체（Calcitionin receptor， CTR）급Notch파기인HES-1、HEY-1 mRNA적표체，TRAP염색감정파골세포，소묘전경검측파골세포용골공능。면역형광검측NICD적표체변화，CCK-8검측RAW 264창7세포증식。결과：Jagged1조TRAP、CK、CTR 급HES-1、HEY-1 mRNA 적표체、TRAP＋세포수교대조조급DAPT조명현승고（P＜0창05），이DAPT조여대조조균무명현변화；세포면역형광현시Jagged1조NICD제표체우세포막、세포질외，세포핵야유교고표체，대조조급DAPT조세포핵중NICD무명현표체；세포배양48 h，Jagged1조세증식교대조조급DAPT조출현명현억제。결론：Jagged1통과활화Notch통로촉진RAW 264창7향파골세포분화、억제기증식。
Objective:To study the role of Jagged1 and Notch signaling pathway played in the differentiation and proliferation of RAW 264.7 cells.Methods: RAW 264.7 cells were divided into three groups to culture:The control group:RAW 264.7 cells were threated with culture and RANKL.The Jagged1 group:RAW 264.7 cells were threated with recombinant protein Jagged 1 besides the control group.The DAPT group:RAW 264.7 cells were threated with DAPT besides the Jagged 1 group.The mRNA expression of osteoclast markers(TRAP,CK,CTR) and Notch key target genes (HES-1 and HEY-1) were measured by real-time PCR.The formation of osteoclast , bone resorption , Notch expression and proliferation of RAW 264.7 cells were detected by TRAP staining , scanning electron microscope ,immunofluorescence and cell counting kit-8 ( CCK-8 ).Results: TRAP, CK, CTR , HES-1 and HEY-1 mRNA expression were significantly higher than the control group and DAPT group in Jadded 1 group ( P<0.05 ).TRAP+cell count ,osteolytic area was significantly increased in Jagged 1 group compared with control and DAPT group , and no significant difference observed between the last two groups.Immunofluorescence results showed high expression of N ICD in cell membrane and cytoplasm in all groups and additionally expressed in nucleus in Jadded 1 group.Cell proliferation was inhibited in Jagged 1 group also ( P<0.05 ).Conclusion:Jagged1 promotes RAW264.7 cells osteoclast differentiation and inhibits proliferation by activating Notch signaling pathway .