| 晚期糖基化终末产物对破骨细胞分化不同阶段的影响 |
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| 引用本文： | 丁晓倩,胡 赟,罗 丹,唐 宇,李彩玉,郑雷蕾. 晚期糖基化终末产物对破骨细胞分化不同阶段的影响[J]. 南方医科大学学报, 2020, 40(4): 573-579. DOI: 10.12122/j.issn.1673-4254.2020.04.20 |
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| 作者姓名： | 丁晓倩 胡 赟 罗 丹 唐 宇 李彩玉 郑雷蕾 |
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| 摘 要： | 目的 探究晚期糖基化终末产物（AGEs）对破骨细胞分化不同阶段的影响。方法 利用核因子κB受体活化因子配体（RANKL）于体外诱导小鼠巨噬细胞系Raw264.7细胞进行破骨细胞向分化，通过抗酒石酸酸性磷酸酶（TRAP）染色确定破骨细胞分化的不同阶段，并将细胞随机分为对照组、分化早期阶段干预组、分化晚期阶段干预组；用CCK-8法检测AGEs作用下的细胞活性；破骨细胞向诱导完成后进行TRAP染色，以RT-PCR 检测RANK、NFATC-1、TRAF-6、TRAP、CTSK的mRNA表达，以Western blot检测CTSK及RANK的蛋白表达。结果 将Raw264.7细胞破骨细胞向分化的前3 d划分为细胞分化早期阶段，之后为细胞分化晚期阶段；100 mg/L的AGEs对Raw264.7细胞的生长无明显影响；早期阶段干预组及晚期阶段干预组的TRAP染色阳性细胞数量均多于对照组，早期阶段干预组与对照组的差异具有统计学意义（P<0.05）；早期阶段干预组及晚期阶段干预组的RANK、NFATC-1、TRAF-6、TRAP、CTSK的mRNA表达均多于对照组，早期阶段干预组与对照组的差异具有统计学意义（P< 0.05）；CTSK及RANK的蛋白表达趋势与相关mRNA表达变化趋势一致。结论 AGEs可能对破骨细胞分化的不同阶段产生不同影响，AGEs于破骨细胞分化早期阶段加入对其分化的促进作用比在晚期阶段加入更明显。
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Effects of advanced glycation end products on osteoclasts at different stages of differentiation |
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| Abstract: | Objective To explore the effect of advanced glycation end products (AGEs) on osteoclasts at different stages ofdifferentiation. Methods Raw264.7 cells cultured in vitro were induced for osteoclastogenesis using RANKL, and the stages ofdifferentiation of the osteoclasts were determined with TRAP staining. The cells were then randomly divided into controlgroup, early-stage AGEs intervention group and late-stage AGEs intervention group. The viability of the cells after AGEstreatment was assessed using CCK-8 method. The cells were examined after the induction for osteoclastogenesis using TRAPstaining, and the expression levels of RANK, NFATC-1, TRAF-6, TRAP and CTSK mRNAs were tested with RT-PCR; theexpressions of CTSK and RANK proteins were detected using Western boltting. Results We defined the initial 3 days ofinduction as the early stage of differentiation and the time beyond 3 days as the late stage of differentiation of Raw264.7 cells.Intervention with AGEs at 100 mg/L produced no significant effects on the viability of the cells, but AGEs suppressed the cellproliferation at a concentration exceeding 100 mg/L. The number of osteolasts in the early- and late-stage intervention groupswas greater than that in the control group, but the cell count differed significantly only between the early-stage interventiongroup and control group (P<0.05). The gene expressions of RANK, NFATC-1, TRAF-6, TRAP and CTSK all increased after theapplication of AGEs in both the early and late stages of differentiation, but the changes were significant only in the early-stageintervention group (P<0.05). The changes in CTSK and RANK protein expressions were consistent with their mRNAexpressions. Conclusion AGEs can affect the differentiation of osteoclasts differently when applied at different stages, andintervention with AGEs at the early stage produces stronger effect to promote osteoclast differentiation than its application at alate stage. |
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