摘要
目的探讨细胞外基质硬化对血管平滑肌细胞(VSMC)迁移的影响.方法采用病理和生理硬度细胞外基质中培养的人冠状动脉平滑肌细胞(HCASMC)的RNA-seq数据,通过生物信息学分析差异表达的基因和富集通路.采用Western blot和qRT-PCR验证病理和生理硬度细胞外基质对HCASMC中磷脂酶C-β2(PLCB2)表达的影响.采用平板划线和小鼠颈动脉内膜损伤模型,验证PLCB2对HCASMC迁移能力的影响.结果差异表达的基因在Gβγ信号通路(P<0.001,FDR=0.17)和PLC-β介导Gβγ信号通路(P<0.001,FDR=0.12)中显著富集,其中PLCB2基因显著上调.PCR[(4.95±0.27),P<0.001]和Western blot[(2.96±0.13),P<0.001].病理性硬化基质培养的HCASMC中PLCB2显著上调.干扰敲低PLCB2能够抑制细胞外基质硬化对HCASMC的促迁移作用.结论病理性细胞外基质硬化通过上调PLCB2促进VSMC迁移.
Objective To investigate the effect of extraceDular matrix stiffness on vascular smooth muscle cell migration.Methods Differentially expressed genes and enrichment pathways were analyzed by bioinfomiatics using RNA-seq data of human coronary arterial smooth muscle cells(HCASMC)cultured in extracellular matrix of pathological and physiological stiffiness.Western blot and qRT-PCR were used to verify the effect of extracellular matrix of pathological and physiological stiffiiess on the expression of PLCB2 in HCASMC.The effects of PLCB2 on the migration of HCASMC were verified by scratch wound assay and mouse carotid intimal injury model.Results Differentially expressed genes were significantly enriched in the Gp y signaling pathway(Nominal p-value<0.001,FDR=0.17)and PLC-β-mediated Gβγsignaling pathway(Nominal p-value<0.001,FDR=0.12),of which PLCB2 was significantly upregulated.PCR[(4.95±0.27),P<0.001]and Western blot[(2.96±0.13),P<0.001]showed that PLCB2 was significantly lip—regulated in HCASMC cultured in pathological stiffness matrix.Scratch wound assay and mouse carotid intimal iiijury model studies found that interference knockdown of PLCB2 inhib让the migration of HCASMC inducing by pathological extracellular matrix stiffness.Conclusion Pathologic al extracellular matrix stiffiiess promotes vascular smooth muscle ceD migration by up-regulating PLCB2.
出处
《浙江临床医学》
2019年第12期1600-1602,共3页
Zhejiang Clinical Medical Journal
基金
浙江省医药卫生科研项目(2017KY559)。
