中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2789-2791
,共3页
Runx2基因%基因沉默%过表达%糖尿病合并骨质疏松
Runx2基因%基因沉默%過錶達%糖尿病閤併骨質疏鬆
Runx2기인%기인침묵%과표체%당뇨병합병골질소송
Runx2 gene%Gene silencing%Overexpression%Diabetes mellitus with osteoporosis
目的 Runx2基因在糖尿病合并骨质疏松患者成骨细胞分化,软骨细胞成熟中的促进作用.方法 构建pLko.1-shRunx2-puro以及pWPI-Runx2-绿色荧光蛋白(GFP)质粒,通过序列测定和Western blot验证这两个质粒的正确性.通过酶联免疫吸附试验(ELISA)法检测Runx2基因沉默和Runx2过表达后,肿瘤坏死因子(TNF)-α和血管内皮生长因子(VEGF)水平变化,探讨Runx2在成骨细胞分化和软骨细胞成熟中的作用.结果 pLko.1-shRunx2-puro以及pWPI-Runx2-GFP质粒构建成功.真核表达后Runx2基因正常组,沉默组和过表达组VEGF的表达水平分别为373.78、10.94、4 161.54 μg/L;而3组TNF-α表达水平分别为62.04、0.22、615.08 ng/L.两者表达水平随着随着Runx2基因的沉默和过表达而降低和升高.结论 在糖尿病合并骨质疏松患者破骨细胞修复过程中,Wnt/β-环连蛋白(β-catenin)信号通路通过激活Runx2表达调节骨质形成.
目的 Runx2基因在糖尿病閤併骨質疏鬆患者成骨細胞分化,軟骨細胞成熟中的促進作用.方法 構建pLko.1-shRunx2-puro以及pWPI-Runx2-綠色熒光蛋白(GFP)質粒,通過序列測定和Western blot驗證這兩箇質粒的正確性.通過酶聯免疫吸附試驗(ELISA)法檢測Runx2基因沉默和Runx2過錶達後,腫瘤壞死因子(TNF)-α和血管內皮生長因子(VEGF)水平變化,探討Runx2在成骨細胞分化和軟骨細胞成熟中的作用.結果 pLko.1-shRunx2-puro以及pWPI-Runx2-GFP質粒構建成功.真覈錶達後Runx2基因正常組,沉默組和過錶達組VEGF的錶達水平分彆為373.78、10.94、4 161.54 μg/L;而3組TNF-α錶達水平分彆為62.04、0.22、615.08 ng/L.兩者錶達水平隨著隨著Runx2基因的沉默和過錶達而降低和升高.結論 在糖尿病閤併骨質疏鬆患者破骨細胞脩複過程中,Wnt/β-環連蛋白(β-catenin)信號通路通過激活Runx2錶達調節骨質形成.
목적 Runx2기인재당뇨병합병골질소송환자성골세포분화,연골세포성숙중적촉진작용.방법 구건pLko.1-shRunx2-puro이급pWPI-Runx2-록색형광단백(GFP)질립,통과서렬측정화Western blot험증저량개질립적정학성.통과매련면역흡부시험(ELISA)법검측Runx2기인침묵화Runx2과표체후,종류배사인자(TNF)-α화혈관내피생장인자(VEGF)수평변화,탐토Runx2재성골세포분화화연골세포성숙중적작용.결과 pLko.1-shRunx2-puro이급pWPI-Runx2-GFP질립구건성공.진핵표체후Runx2기인정상조,침묵조화과표체조VEGF적표체수평분별위373.78、10.94、4 161.54 μg/L;이3조TNF-α표체수평분별위62.04、0.22、615.08 ng/L.량자표체수평수착수착Runx2기인적침묵화과표체이강저화승고.결론 재당뇨병합병골질소송환자파골세포수복과정중,Wnt/β-배련단백(β-catenin)신호통로통과격활Runx2표체조절골질형성.
Objective To investigate the promoting role of Runx2 gene in the osteoblasts and chondrocytes maturation of diabetes mellitus patients with osteoporosis.Methods Constructed pLko.1-shRunx2-puro and pWPI-Runx2-green fluorescent protein(GFP) plasmids and verify the correctness of the two plasmids by sequencing and Western blotting.Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor (TNF)-α and vascular endothelial growth factor (VEGF) after overexpression and silencing of Runx2 gene, in order to explore the role of Runx2 in osteoblast differentiation and maturation of chondrocytes.Results pLko.1-shRunx2-puro and pWPI-Runx2-GFP plasmid was successfully constructed.After eukaryotic expression, the VEGF expression levels in the Runx2 normal expression group, the Runx2 silencing expression group and the Runx2 overexpression group were 373.78, 10.94, 4 161.54 μg/L, respectively;and the TNF-α expression levels were 62.04, 0.22,615.08 ng/L, respectively.The expression levels of VEGF and TNF-o expression levels lowered and raised as the decrease and increase of Runx2 gene.Conclusion In osteoclasts repair process of diabetes mellitus patients with osteoporosis, Wnt/β-catenin signaling pathway regulated bone formation by activating the expression of Runx2 gene.
