·糖尿病基础研究·

　　磺脲类药物对成骨细胞MC3T3-E1自噬、凋亡和功能的影响

　　张丽 季虹 苏华 刘兴艳 辛衍代 荣海钦

　　【摘要】 目的 探讨磺脲类药物对成骨细胞自噬、凋亡及其功能的影响。 方法 用磺酰罗丹明B染色检测不同浓度的格列本脲(GLB)、格列齐特(GLC)和格列吡嗪(GLP)对成骨细胞存活率的影响;Western blot分析三种磺脲类药物对细胞中自噬、凋亡标志蛋白表达的变化;Hoechst染色镜下观察上述药物对细胞凋亡的影响;通过对骨钙素(OCN)和碱性磷酸酶(ALP)的测定研究药物对细胞分化功能的影响。 结果 中、高浓度的GLB、GLC和GLP使MC3T3-E1细胞存活率降低。在药物干预下，MC3T3-E1细胞自噬和凋亡标志蛋白表达增加，mTOR信号途径无明显变化。三种药物可使细胞分泌OCN和ALP的能力下降。 结论 GLB、GLC和GLP可诱导成骨细胞MC3T3-E1发生自噬和凋亡，降低成骨细胞分化功能，且可能通过mTOR-非依赖途径诱导细胞自噬。

　　【关键词】 磺脲药物;成骨细胞;自噬;凋亡;分化功能

　　Influence of sulfonylureas on autophagy, apoptosis, and differentiation of the mice MC3T3-E1 cells ZHANG Li, JI Hong, SU Hua, et al. Shandong Institute of Endocrine and Metabolic Diseases, Jinan 250062, China

　　Corresponding author: RONG Hai-qin, E-mail: haiqinrong@126.com

　　【Abstract】Objective To investigate the influence of sulfonylureas on the autophagy, apoptosis, and differentiation of the MC3T3-E1 cells. Methods Sulforhodamine B Assay was carried out to determine the effect of glibenclamide, gliclazide and glipizide in different concentrations on the cell survival. Western blot analysis was done to determine the protein levels of autophagy and apoptosis markers. Hoechst staining was applied to observe the impact of the three sulfonylureas on the cell apoptosis. The osteocalcin and alkaline phosphatase were assayed to assess the influence of these drugs on the differentiation of MC3T3-E1 cells. Results The intervention with glibenclamide, gliclazide, and glipizide in medium and high concentrations made the survival rate of MC3T3-E1 cells decreased. Under the condition of drug intervention, the expression of autophagy and apoptosis markers of MC3T3-E1 cells was enhanced, whereas no remarkable changes were detected in mTOR signaling. The three sulfonylureas weakened the secretion of osteocalcin and alkaline phosphatase of MC3T3-E1 cells. Conclusion Glbenclamide, gliclazide, and glipizide induce the autophagy and apoptosis of MC3T3-E1 cells, decrease the osteoblastic differentiation, and may induce autophagy via an mTOR-independent pathway.

　　【Key Words】 Sulfonylureas; Osteoblast; Autophagy; Apoptosis; Differential function