摘要
目的探讨糖基化终末产物（AGEs）对间充质干细胞（MSCs）增殖和迁移能力的影响及其机制。方法体外培养SD大鼠MSCs，流式细胞术鉴定阳性指标（CD29和CD90）和阴性指标（CD34和CD45）。噻唑蓝（Myr）比色法检测不同剂量（0、25、50、100、200mg／L）AGEs-牛血清白蛋白（BSA）在刺激不同时间（6、12、24h）后MSCs增殖变化。采用Wound—healing划痕实验检测不同剂量（0、25、50、100、200mg／L）AGEs—BSA刺激骨髓间充质干细胞24h后的迁移能力。荧光染色拍照测定AGEs—BSA（200mg／L）刺激12、24h后细胞内活性氧（ROS）累积情况。结果200mg／LAGEs—BSA刺激6、12、24hMSCs增殖能力显著下降，分别为81．22％、80．53％、51．50％，AGEs—BSA抑制细胞增殖呈时间和剂量依赖性。200mg／LAGEs—BSA刺激24h迁移能力下降[（0．012±0．006）μm／min比（0．022±0．002）μm／min，P〈0．01]。同时，细胞内的ROS在AGEs—BSA刺激下大量累积。结论AGEs—BSA刺激下通过其受体导致细胞内ROS累积，从而抑制细胞增殖和迁移能力。
Objective To explore the mechanism of advanced glycation end products （AGEs） attenuating the proliferation and migration in mesenchymal stem cells （MSCs）. Methods The MSCs were isolated from male Sprague Dawley rats, and identified by flow cytometry （ positive ： CD29 and CD90 ; negative： CD34 and CIM5）. MSCs were stimulated by AGEs -bovine serum albumin （BSA） with different doses （0, 25, 50, 100 and 200 mg/L） and time （6, 12 and 24 h）. The proliferation of cells was examined by methyl thiazol tetrazolium （MTF） assay. Wound - healing assay was used to assess the migration potential of cells treated with AGEs - BSA. Intracellular formation of reactive oxygen species （ROS） was evaluaed using a fluorescent probe. Results The relative metabolic activity was significantly reduced by 81.22%, 80. 53% and 51.50% with AGEs - BSA （200 mg/L） at 6, 12, and 24 h, respectively. The proliferation potential of MSCs was impacted by AGEs - BSA in a dose - and time - dependent manner. AGEs - BSA （200 mg/L for 24 h） obviously inhibited migration of MSCs [ （0. 012 ±0. 006） μm/min vs. （0. 022 ±0. 002） μm/min,P 〈0.01 ]. At the same time, AGEs - BSA significantly induced the intracellular ROS accumulation. Conclusion AGEs - BSA induced ROS formation via receptor to inhibit proliferation and migration.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第2期436-438,共3页
Chinese Journal of Experimental Surgery
关键词
糖基化终末产物
骨髓间充质干细胞
增殖
迁移
Advanced glycation end products
Bone mesenchymal stem cells
Proliferation
Migration
