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目的探讨磺脲类药物对成骨细胞自噬、凋亡及分化功能的影响。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3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。3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。
Objective To investigate the effects of sulfonylureas on autophagy, apoptosis and differentiation of osteoblasts. Methods The effects of different concentrations of GLB, GLC and GLP on the viability of osteoblasts were detected by sulforhodamine B staining. The changes of autophagy and apoptosis marker proteins in ureaplasma were observed. The effects of the above drugs on apoptosis were observed by Hoechst staining. The effects of these drugs on apoptosis were analyzed by measuring the levels of osteocalcin (OCN) and alkaline phosphatase (ALP) Effect of drugs on cell differentiation. As a result, high concentrations of GLB, GLC and GLP decreased the survival rate of MC3T3-E1 cells. Under drug intervention, the expression of autophagy and apoptosis marker protein increased in MC3T3-E1 cells, but there was no obvious change in mTOR signaling pathway. The three drugs decreased the ability of cells to secrete OCN and ALP. Conclusion GLB, GLC and GLP can induce autophagy and apoptosis of osteoblast MC3T3-E1, reduce osteoblast differentiation and induce autophagy via mTOR-independent pathway.