CAJ | 학술논문

目的：观察高铁培养环境下小鼠前成骨样细胞MC3T3-E1增殖、分化指标的变化趋势，探讨铁离子对成骨细胞增殖、分化的影响。方法小鼠前成骨样细胞MC3T3-E1在37℃条件下体外培养，在10 mmol/Lβ-甘油磷酸和50μg /mL抗坏血酸的诱导分化的作用下，分化为成骨细胞，同时用不同浓度（50、100、200μmol/L）枸橼酸铁铵（ FAC）干预，用MTT法检测细胞的增殖活性，RT-PCR法检测成骨细胞分化基因成骨相关转录因子（Runx2）、锌指结构转录因子（Osterix）、骨唾液酸蛋白（BSP）和骨钙素（OC）的表达，碱性磷酸酶（ALP）活性试剂盒检测细胞碱性磷酸酶活性。结果 MC3T3-E1细胞的增殖活性、成骨分化相关基因的表达以及ALP水平随FAC干预浓度的增加呈剂量依赖性降低（ P＜0.05）。结论高铁培养环境可明显抑制小鼠前成骨样细胞MC3T3-E1的增殖和分化。
목적：관찰고철배양배경하소서전성골양세포MC3T3-E1증식、분화지표적변화추세，탐토철리자대성골세포증식、분화적영향。방법소서전성골양세포MC3T3-E1재37℃조건하체외배양，재10 mmol/Lβ-감유린산화50μg /mL항배혈산적유도분화적작용하，분화위성골세포，동시용불동농도（50、100、200μmol/L）구연산철안（ FAC）간예，용MTT법검측세포적증식활성，RT-PCR법검측성골세포분화기인성골상관전록인자（Runx2）、자지결구전록인자（Osterix）、골타액산단백（BSP）화골개소（OC）적표체，감성린산매（ALP）활성시제합검측세포감성린산매활성。결과 MC3T3-E1세포적증식활성、성골분화상관기인적표체이급ALP수평수FAC간예농도적증가정제량의뢰성강저（ P＜0.05）。결론고철배양배경가명현억제소서전성골양세포MC3T3-E1적증식화분화。
Objective To observe the changing pattern of proliferative and differentiatial indexes of mouse MC 3T3-E1 preosteoblasts in the culture with excess iron , and to investigate the effect of iron on the proliferation and differentiation of osteoblasts .Methods Mouse MC3T3-E1 preosteoblasts were cultured at 37℃in vitro.Under the induction of 10mmol/lβ-glycerophosphate and 50μg/ml L-ascorbic acid, MC3T3-E1 preosteoblasts differentiated into osteoblasts .Meanwhile, different concentrations (50, 100, and 200μmol/l) of FAC were additioned into the medium for intervention .Proliferation of MC3T3-E1 cells was detected using MTT method .The mRNA expressions of Runx2, Osterix, bone sialoprotein (BSP), and osteocalcin (OC) were detected using RT-PCR.ALP activity was measured using ALP viability kit .Results The proliferation of MC3T3-E1 cells, the mRNA expressions of Runx2, Osterix, BSP, and OC, and ALP activity decreased along with the increase of FAC concentration , showing a dose-dependent manner ( P<0.05). Conclusion Excess iron culturing can significantly inhibit the proliferation and differentiation of mouse MC 3T3-E1 preosteoblasts.