中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
32期
2288-2291
,共4页
郭英%李佩芳%舒晓春%邓欢%马海丽%孙辽
郭英%李珮芳%舒曉春%鄧歡%馬海麗%孫遼
곽영%리패방%서효춘%산환%마해려%손료
骨质疏松%骨碎补总黄酮%骨髓间充质干细胞%成骨分化
骨質疏鬆%骨碎補總黃酮%骨髓間充質榦細胞%成骨分化
골질소송%골쇄보총황동%골수간충질간세포%성골분화
Osteoporosis%Drynaria total flavonoids%Bone marrow mesenchymal stem cells%Osteogenic differentiation
目的 探讨骨碎补总黄酮对骨髓间充质干细胞(BMSC)成骨分化过程中,对Wnt/β-catenin信号通路相关因子mRNA表达的影响.方法 采用全骨髓贴壁法分离、培养SD大鼠BMSC,取3代细胞予100μg/ml浓度骨碎补总黄酮进行干预,21 d后,观察并行矿化结节染色,在干预7、14、21、28 d时检测成骨分化标志物碱性磷酸酶(ALP)活性和Wnt/β-catenin信号通路相关因子β-catenin、淋巴增强因子1(LEF-1)、细胞周期蛋白D(cycline D) mRNA的表达情况.结果 干预后各个时间点骨碎补总黄酮组ALP活性(U/L)均较空白组高(7 d:11.10±0.08比1.61±0.14；14 d:24.62±0.34比1.64±0.04;21 d:18.4 ±0.06比1.53 ±0.04；28 d:14.91 ±0.14比1.52 ±0.04;均P ＜0.01)；21 d后行矿化结节染色,骨碎补总黄酮组为阳性,空白组为阴性；干预后β-catenin mRNA在第14天表达最高(0.357±0.062比0.174 ±0.013,P＜0.05)；LEF-1、cyclineD mRNA在第7大表达最高(LEF-1:0.0611 ±0.0002比0.0345±0.0131；cycline D:0.1510±0.0255比0.0718±0.0294均P＜0.05).结论 骨碎补总黄酮诱导大鼠BMSC向成骨细胞分化,伴随Wnt/β-catenin信号通路相关因子mRNA表达的变化.
目的 探討骨碎補總黃酮對骨髓間充質榦細胞(BMSC)成骨分化過程中,對Wnt/β-catenin信號通路相關因子mRNA錶達的影響.方法 採用全骨髓貼壁法分離、培養SD大鼠BMSC,取3代細胞予100μg/ml濃度骨碎補總黃酮進行榦預,21 d後,觀察併行礦化結節染色,在榦預7、14、21、28 d時檢測成骨分化標誌物堿性燐痠酶(ALP)活性和Wnt/β-catenin信號通路相關因子β-catenin、淋巴增彊因子1(LEF-1)、細胞週期蛋白D(cycline D) mRNA的錶達情況.結果 榦預後各箇時間點骨碎補總黃酮組ALP活性(U/L)均較空白組高(7 d:11.10±0.08比1.61±0.14；14 d:24.62±0.34比1.64±0.04;21 d:18.4 ±0.06比1.53 ±0.04；28 d:14.91 ±0.14比1.52 ±0.04;均P ＜0.01)；21 d後行礦化結節染色,骨碎補總黃酮組為暘性,空白組為陰性；榦預後β-catenin mRNA在第14天錶達最高(0.357±0.062比0.174 ±0.013,P＜0.05)；LEF-1、cyclineD mRNA在第7大錶達最高(LEF-1:0.0611 ±0.0002比0.0345±0.0131；cycline D:0.1510±0.0255比0.0718±0.0294均P＜0.05).結論 骨碎補總黃酮誘導大鼠BMSC嚮成骨細胞分化,伴隨Wnt/β-catenin信號通路相關因子mRNA錶達的變化.
목적 탐토골쇄보총황동대골수간충질간세포(BMSC)성골분화과정중,대Wnt/β-catenin신호통로상관인자mRNA표체적영향.방법 채용전골수첩벽법분리、배양SD대서BMSC,취3대세포여100μg/ml농도골쇄보총황동진행간예,21 d후,관찰병행광화결절염색,재간예7、14、21、28 d시검측성골분화표지물감성린산매(ALP)활성화Wnt/β-catenin신호통로상관인자β-catenin、림파증강인자1(LEF-1)、세포주기단백D(cycline D) mRNA적표체정황.결과 간예후각개시간점골쇄보총황동조ALP활성(U/L)균교공백조고(7 d:11.10±0.08비1.61±0.14；14 d:24.62±0.34비1.64±0.04;21 d:18.4 ±0.06비1.53 ±0.04；28 d:14.91 ±0.14비1.52 ±0.04;균P ＜0.01)；21 d후행광화결절염색,골쇄보총황동조위양성,공백조위음성；간예후β-catenin mRNA재제14천표체최고(0.357±0.062비0.174 ±0.013,P＜0.05)；LEF-1、cyclineD mRNA재제7대표체최고(LEF-1:0.0611 ±0.0002비0.0345±0.0131；cycline D:0.1510±0.0255비0.0718±0.0294균P＜0.05).결론 골쇄보총황동유도대서BMSC향성골세포분화,반수Wnt/β-catenin신호통로상관인자mRNA표체적변화.
Objective To explore the effects of the expression of Wnt/β-catenin signaling factor mRNA during drynaria total flavonoids on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).Methods The BMSCs were isolated from SD rats by whole bone marrow culture method and purified by passage.And the P3 BMSCs were intervened with 100 μg/ml drynaria total flavonoids.At Day 21,mineralized staining was performed. At Days 7,14,21 and 28 post-intervention,the activity ofalkaline phosphatase (ALP) was detected and polymerase chain reaction (PCR) used to detect theexpressions of Wnt/β-catenin signaling pathway related factors β-catenin,LEF-1 and cycline D mRNA.Results At each timepoint post-intervention,comparing the ALP activity in cell supematant between two group,the drynaria total flavonoids group was higher than the blank control group (7 d:11.10 ± 0.08 vs 1.61 ±0.14; 14 d:24.62±0.34 vs 1.64±0.04; 21 d:18.41 ±0.06 vs 1.53 ±0.04; 28 d:14.9 ±0.14vs 1.52 ±0.04; all P ＜ 0.01 ).At Day 21,upon staining with alizarin red,the drynaria total flavonoids group was positive while the blank control group negative.At Day 14,the expression of β-catenin mRNA was higher in the drynaria total flavonoids group higher than that in the blank control group (0.357 ±0.063 vs 0.174 ±0.013,P ＜0.05).At Day 7,the expressions of LEF-1 and cycline D mRNA were higher in the drynaria total flavonoids group than those in the blank control group ( LEF-1 0.0611 ± 0.0002 vs 0.0345 ±0.0131 ; cycline D 0.1510 ± 0.0255 vs 0.0718 ± 0.0294,all P ＜ 0.05 ).Conclusion Drynaria total flavonoids induce BMSCs to differentiate into osteoblasts.And it is accompanied with the altered expression of Wnt/β-catenin signaling pathway related factor mRNA.
