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目的观察高糖环境下吡格列酮对MC3T3-E1成骨细胞的作用并探讨其可能机制。方法高糖(22.5 mmol·L~(-1))环境培养MC3T3-E1细胞,分为对照组,吡格列酮2.5、5、10μmol·L~(-1)组,干预24、48 h。检测细胞增殖活性、凋亡率、骨钙素和碱性磷酸酶(ALP)分泌水平,以及过氧化物酶体增殖物激活受体γ(PPARγ)、成骨因子runt相关基因2(Runx2)和骨形态蛋白2(BMP-2)mRNA的表达水平。并分析PPARγ、Runx2的表达与骨钙素、ALP、BMP-2的相关性。结果在相同干预时限,吡格列酮各组MC3T3-E1成骨细胞增殖活性、骨钙素和ALP的分泌水平、Runx2 mRNA和BMP-2 mRNA的表达均低于对照组,凋亡率和PPARγmRNA的表达高于对照组(P<0.05)。随吡格列酮浓度增加,细胞增殖活性、骨钙素和ALP的分泌、Runx2 mRNA和BMP-2 mRNA的表达均降低,而细胞凋亡率和PPARγmRNA的表达增高(P<0.05)。与干预24 h相比,干预48 h时相同浓度吡格列酮组细胞增殖活性,骨钙素和ALP的分泌水平,PPARγ、Runx2、BMP-2 mRNA的表达或无变化或略增加。结论高糖环境下吡格列酮对成骨细胞有损害作用,促进PPARγ表达、抑制Runx2的表达可能为其作用机制之一。
Objective To observe the effect of pioglitazone on MC3T3-E1 osteoblasts under high glucose conditions and to explore its possible mechanism. Methods MC3T3-E1 cells were cultured in high glucose (22.5 mmol·L -1) environment and divided into control group, 2.5, 5 and 10 μmol·L -1 pioglitazone groups for 24 and 48 h. The cell proliferation activity, apoptosis rate, osteocalcin and alkaline phosphatase (ALP) secretion levels, as well as peroxisome proliferator activated receptor γ (PPARγ), osteoblast ruun2 (Runx2) and Bone morphogenetic protein 2 (BMP-2) mRNA expression levels. The correlation between PPARγ, Runx2 expression and osteocalcin, ALP and BMP-2 was analyzed. Results In the same intervention time period, the proliferation activity, the secretion of osteocalcin and ALP, the expression of Runx2 mRNA and BMP-2 mRNA of MC3T3-E1 in each group were lower than those in the control group, and the apoptosis rate and PPARγ mRNA expression were high In the control group (P <0.05). With the increase of pioglitazone concentration, the cell proliferation activity, the secretion of osteocalcin and ALP, the expression of Runx2 mRNA and BMP-2 mRNA decreased, but the apoptosis rate and PPARγ mRNA expression increased (P <0.05). Compared with the 24 h intervention, the cell proliferation activity, the secretion of osteocalcin and ALP, the expression of PPARγ, Runx2 and BMP-2 mRNA in the same concentration of pioglitazone group at 48 h were unchanged or slightly increased. Conclusion Pioglitazone has a detrimental effect on osteoblasts in high glucose environment and promotes the expression of PPARγ. Inhibition of Runx2 may be one of its mechanisms.