过表达SCD1对骨髓间质干细胞成骨分化的影响及其基因表达谱变化研究

目的:研究固醇辅酶A去饱和酶1(SCD1)过表达后对骨髓间质干细胞(BM-MSCs)成骨分化作用的影响,并利用基因芯片技术分析基因表达谱的变化。方法:利用已构建成功的SCD1慢病毒转染BM-MSCs,采用RT-PCR及C14技术检测SCD1在BM-MSCs中过表达情况及其活性。成骨诱导培养BM-MSCs后,采用Western blot和茜素红染色技术检测骨钙素(OC)等相关成骨指标,进一步运用全基因芯片检测过表达SCD1对BM-MSCs成骨分化表达谱的影响。结果:SCD1在BM-MSCs中成功过表达,过表达组SCD1活性明显高于对照组。成骨诱导7天、14天时,过表达组中的碱性磷酸酶(APL)活性和骨钙素水平均明显高于对照组(P<0.05)。成骨诱导一周、两周时,过表达组的碱性磷酸酶染色和茜素红染色均多于对照组。基因表达芯片的结果显示,过表达SCD1改变骨髓间质干细胞表达谱,检测出差异基因2896个。基因通路分析提示干扰素通路为表达差异最显著通路(P<0.05)。结论:过表达SCD1可以促进BM-MSCs的成骨分化,可能通过作用于干扰素通路影响成骨分化功能。这一发现可能为骨折愈合提供重要的思路和潜在治疗策略,值得深入研究。

Effect of SCD1 Overexpression on the Osteogenic Differentiation Function of Bone Marrow Mesenchymal Stem Cells and Differential Gene Expression Profile

ABSTRACT Objective: To investigate the effect of steroid coenzyme A desaturase 1 (SCD1) overexpression on osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs), and to analyze the changes of whole genome expression profile by gene chip technology. Methods: BM-MSCs were transfected with SCD1 lentivirus. The overexpression and activity of SCD1 in BM-MSCs were detected by RT-PCR and C14 technique. OC and other related osteogenesis indexes were detected by Western blot and alizarin red staining after BM-MSCs were induced and cultured. The effect of overexpression of SCD1 on osteogenic differentiation expression profile of BM-MSCs was further examined by whole gene chip. Results: SCD1 was successfully overexpressed in BM-MSCs, and SCD1 activity in overexpressing group was significantly higher than that in control group. The levels of alkaline phosphatase (APL) and osteocalcin (OC) in the overexpression group were higher than those in the control group at 7 and 14 days after osteogenesis induction (P < 0.05). Alkaline phosphatase staining and alizarin red staining were more in the overexpression group than in the control group at 7 and 14 days after osteogenesis induction. The results of gene expression microarray showed that 2896 differentially expressed genes were detected by overexpression of SCD1 in bone marrow mesenchymal stem cells. Gene pathway analysis showed that the interferon pathway was the most significant pathway (P<0.05). Conclusion: Overexpression of SCD1 can promote the osteogenic differentiation of BM-MSCs, which may affect the osteogenic differentiation function through interferon pathway. This finding may provide important thinking and potential therapeutic strategies for fracture healing, and is worthy of further study.