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目的:研究血红素加氧酶1(hemeoxygenase 1,Hmox1)在骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9)诱导下间充质干细胞C3H10T1/2向成骨分化过程中发挥的作用。方法:用Ad-BMP9感染C3H10T1/2,分别用Q-PCR和Western blot检测Hmox1mRNA和蛋白水平的变化;Hmox1激动剂COPP处理BMP9诱导的C3H10T1/2细胞,碱性磷酸酶(ALP)染色和活性测定检测早期成骨指标ALP的变化;过表达Hmox1的重组腺病毒(Ad-Hmox1)处理BMP9诱导的C3H10T1/2细胞,ALP染色和活性测定早期成骨指标ALP,茜素红染色检测晚期成骨指标钙盐沉积,Western blot检测成骨相关基因COL1A1。结果:Ad-BMP9感染C3H10T1/2后,Hmox1的mRNA及蛋白水平均升高;BMP9与Hmox1激动剂COPP联用与BMP9组相比ALP的活性增强;Ad-Hmox1可以增强BMP9诱导下C3H10T1/2细胞的ALP活性和钙盐沉积,以及成骨相关基因COL1A1表达。结论:Hmox1可以促进BMP9诱导下C3H10T1/2细胞的成骨分化。
AIM: To investigate the role of hemeoxygenase 1 (Hmox1) in the osteogenic differentiation of mesenchymal stem cells (C3H10T1 / 2) induced by bone morphogenetic proteins 9 (BMP9). Methods: C3H10T1 / 2 cells were infected with Ad-BMP9, and the mRNA and protein levels of Hmox1 were detected by Q-PCR and Western blot respectively. H9 cells treated with BMP9 were treated with Hmox1 agonist COPP for 3 hours, and alkaline phosphatase (ALP) The expression of ALP in early osteoblast was detected by immunohistochemistry and the expression of ALP was detected by ALP staining and ALP staining. The expression of ALP was detected by alizarin red staining in advanced C3H10T1 / 2 cells treated with recombinant adenovirus over-expressing Hmox1 (Ad-Hmox1) Indicators of calcium deposition, Western blot detection of osteogenic related gene COL1A1. RESULTS: The mRNA and protein levels of Hmox1 in C3H10T1 / 2 infected with Ad-BMP9 were all increased. The activity of ALP in BMP9-treated HMCs was higher than that in BMP9 group. Ad-Hmox1 enhanced the expression of C3H10T1 / 2 Cell ALP activity and calcium deposition, and osteogenesis-related gene COL1A1 expression. Conclusion: Hmox1 can promote the osteogenic differentiation of C3H10T1 / 2 cells induced by BMP9.