| 吴宁 卢晓昭 曹晓瑞 于立峰 陶惠人 朱庆生.高浓度葡萄糖通过p38MAPK-TXNIP/TRX-ROS通路抑制MC3T3-E1
细胞成骨分化[J].,2014,14(12):2224-2229 |
| 高浓度葡萄糖通过p38MAPK-TXNIP/TRX-ROS通路抑制MC3T3-E1
细胞成骨分化 |
| High Glucose Suppresses Osteoblastic Differentiation of MC3T3-E1 CellsVia p38 MAPK-TXNIP/TRX-ROS Signal Pathway |
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| DOI： |
| 中文关键词: 糖尿病性骨质疏松症 p38 MAPK 氧化应激 成骨细胞分化 |
| 英文关键词: Diabetic osteoporosis P38 MAPK Oxidate stress Osteoblast differentiation |
| 基金项目:国家自然科学基金项目（81070698） |
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| 中文摘要: |
| 目的：研究高浓度葡萄糖抑制MC3T3-E1细胞成骨分化的机理。方法：建立MC3T3-E1 细胞成骨分化诱导体系，观察不同浓
度葡萄糖（5.5mM 和22mM）对MC3T3-E1 细胞成骨分化的影响；用不同浓度的p38 MAPK 抑制剂Fr167653（0.1 滋M、1.0 滋M 和
10 滋M）进行药物干预，观察MC3T3-E1 细胞在22mM葡萄糖浓度下成骨分化的变化情况。通过钙含量检测、Real time PCR 检测
相关分化的变化；用Western Blot 方法检测MC3T3-E1 细胞分化过程中p38 MAPK 磷酸化状态、TXNIP 表达水平的变化；使用胰
岛素二硫键还原法检测细胞内TRX活性水平；使用活性氧检测试剂盒检测细胞内自由氧生成水平。结果：体外诱导条件下，高浓
度（22mM）葡萄糖通过升高p38 MAPK 磷酸化水平，上调TXNIP 表达水平，同时降低TRX 活性，使细胞内自由氧生成增加，抑制
MC3T3-E1 细胞的成骨分化；Fr167653通过抑制p38 MAPK 磷酸化，下调TXNIP 表达同时升高TRX活性，抑制细胞内自由氧生
成，解除高浓度葡萄糖对细胞成骨分化的抑制作用。结论：高浓度葡萄糖通过p38 MAPK-TXNIP/TRX-ROS 信号通路抑制
MC3T3-E1细胞成骨分化。 |
| 英文摘要: |
| Objective:Our study focus on the role of p38 MAPK and ROS in osteoblast differentiation of
MC3T3-E1 cell under high concentration of glucose in vitro.Methods:MC3T3-E1 cells were divided into four groups: negtive control
group(NC, MC3T3-E1 cells cultured in 琢-MEMmedium), normal glucose and calcification induced control group(NIC, MC3T3-E1 cells
cultured in calcification medium with 5.5mM glucose), high glucose group (HG, MC3T3-E1 cells cultured in calcification medium with
22mMglucose) and Fr167653 intervention group(Fr~, MC3T3-E1 cells cultured in calcification mediumwith 22mMglucose, in the presence
of p38 MAPK inhibitor, Fr167653). The calciumdeposition, alkaline phosphatase activity and the mRNA levels of osteoblast markers
were tested to evaluate the osteoblast differentiation potency of MC3T3-E1 cells. P38 MAPK phosphorylation level and TXNIP protein
expression level were detected by Western blot. Intracellular TRX activity was detected using the insulin disulfide reduction assay
method. Intracellular ROS generation level was detected by Reactive Oxygen Species Assay Kit.Results:We showed in this report that
22mMglucose suppressed the osteoblast differentiation process of MC3T3-E1 cells, as indicated by the reduced calcium deposition, ALP
activity and osteoblast markers. In the presence of Fr167653, the specific inhibitor of p38 MAPK, could significantly reverse the suppression
effect of high glucose and the decrease of above-mentioned osteoblast differentiation markers. Furthermore, high glucose increased
p38 MAPK phosphorylation level. The TXNIP expression level was up-regulated and the TRX activity was decreased, which further increased
the intracellular ROS generation. Fr167653, which blocked the p38 MAPK phosphorylation, down-regulated the TXNIP expression,
enhanced the TRX activity and reduced the ROS generation.Conclusion:These data suggest that high concentration of glucose had
a deleterious effect on osteoblast differentiation of MC3T3-E1 cells probably via p38 MAPK-TXNIP/TRX-ROS signal pathway. |
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