摘要
目的探讨赖氨酸甲基转移酶SET8对大鼠血管平滑肌细胞(VSMCs)向成骨样细胞分化的影响。方法体外分离培养大鼠VSMCs,将VSMCs随机分为正常对照组(未转染)、空质粒组(转染NS-sh RNA)、SET8-sh RNA组。应用反转录聚合酶链反应(RT-PCR)、蛋白免疫印迹(Western blot)法分别检测SET8和调节成骨与软骨分化的关键转录因子RUNX2 m RNA和蛋白的表达水平;应用酶联免疫法检测碱性磷酸酶(ALP)活性。结果 SET8-sh RNA可成功抑制VSMCs中SET8的表达;RT-PCR及Western blot结果显示,SET8-sh RNA组RUNX2 m RNA和蛋白的表达较正常对照组和空质粒组明显下降(P<0.05)。ALP活性测定结果显示,SET8-si RNA组ALP活性较正常对照组和空质粒组显著降低(P<0.05)。结论干扰SET8基因表达能够抑制大鼠VSMCs向成骨样细胞转分化。
Objective To investigate the effect of lysine methyltransferase SET8on regulating cell transdifferentiation of rat vascular smooth muscle cells(VSMCs)into osteoblast-like cells.Methods VSMCs were obtained from rat thoracic aorta,and then randomly divided into control group(non-transfection),the empty plasmid group(transfect NS-shRNA)and SET8-shRNA group.The expression of SET8,runt?related transcription factor2(RUNX2)was detected by RT?PCR and Western blot assay.Alkaline phosphatase(ALP)activity was measured by enzyme linked immunoassay.Results The expression of SET8in VSMCs was effectively inhibited by SET8-shRNA.RT-PCR and Western blot results showed that the expression of RUNX2was decreased in cells after SET8-shRNA transfection(P<0.05).ALP activity was significantly reduced after SET8-shRNA transfection(P<0.05).Conclusion Interference with SET8gene expression could inhibit the differentiation of VSMCs into osteo-like cells.
作者
张东雪
高少辉
李同妙
周薇
陈丽洁
ZHANG Dong-xue;GAO Shao-hui;LI Tong-miao;ZHOU Wei(Departments of Nephrology, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China)
出处
《天津医药》
CAS
北大核心
2018年第3期255-258,共4页
Tianjin Medical Journal
