骨碎补总黄酮在不同糖浓度下对大鼠骨髓间充质干细胞成骨分化的影响  

Effects of drynaria total flavonoid on osteogenic differentiation of bone marrow mesenchymal stem cells at different glucose concentrations： experiment with rats

作　　者：舒晓春[1] 朱丹华[1] 庞天娇[1] 孙辽[1] 叶礼红[1] 鲁红云[1] 尹代婵[1] 谢丹红[1] 

机构地区：[1]中山大学附属第五医院内分泌科,珠海519000

出　　处：《中华医学杂志》2010年第38期2708-2712,共5页National Medical Journal of China

基　　金：国家中医药管理局科研基金（04-05ZP51）;广东省中医药管理局科研基金（2007263）

摘　　要：目的 了解骨碎补总黄酮在不同糖浓度下对骨髓间充质干细胞（BMSC）成骨分化的影响,为在糖尿病性骨质疏松症的防治提供理论依据.方法 对SD大鼠BMSC在体外条件下培养,进行定向成骨诱导分化,采用全骨髓贴壁法分离纯化BMSC.实验分为低糖组、高糖组、低糖诱导组、高糖诱导组、低糖骨碎补组、高糖骨碎补组6组,检测相关成骨指标碱性磷酸酶（ALP）活性、矿化结节数目、胶原蛋白.观察骨碎补总黄酮在不同糖浓度下对BMSC成骨分化的影响.结果 骨碎补总黄酮能促进BMSC成骨指标的表达,低糖骨碎补组成骨指标的表达高于低糖诱导组[ALP活性（0.439±0.024比0.385±0.029）、ALP染色阳性率（48.7%比35.0%）、矿化结节数（9.75±1.71个/HP比6.25 ±0.96 个/HP）、Ⅰ型胶原蛋白积分（2.21±0.07比1.93±0.13）]差异均有统计学意义（均P＜0.05）.高糖骨碎补组高于高糖诱导组[ALP活性（0.352±0.022比0.139±0.013）、ALP染色阳性率（25.3%比22.7%）、矿化结节数（4.50±1.29个/HP比3.25±1.50个/HP）、Ⅰ型胶原积分（1.70±0.03比1.28±0.27）]差异均有统计学意义（均P＜0.05）.不同糖浓度组的糖基化终末产物（AGE）水平随糖浓度和培养时间延长呈增高趋势（均P＜0.05）,与Ⅰ型胶原蛋白的表达呈负相关（r=-0.410,P＜0.05）.结论 骨碎补总黄酮可以促进BMSC成骨分化并减轻高糖条件下对成骨分化的抑制作用.Objective To study the effects of drynaria total flavonoid on osteogenic differentiation of bone marrow mesenchymal stem cells （BMSCs） at different glucose concentrations. Methods BMSCs of SD rats were isolated, cultivated in vitro, and divided into 6 groups to be induced to differentiate into osteoblasts was used to examine the level of ALP. The ALP staining positive rate was examined with modified calcium cobalt method. Alizarin red staining was adopted to observe the number of calcium nodes.Immunohistochemistry was used to detect type Ⅰ collagen level. Advanced glycosylation end products （AGEs） were tested by ELISA. Results The A value indicating the ALP activity, ALP staining positive rate, calcium node number, and type Ⅰ collagen expression score of the low glucose ＋ drynaria total flavonoid group were （0.439 ± 0.024）, 48.7%, （9.75 ± 1.71） nodes/HP, and （2.21 ±0.07）respectively, all significantly higher than those of the sodium glycerophophate ＋ vitamin C ＋ dexamethasone [（0. 385 ± 0. 029）, 35.0%, （6. 25 ± 0. 96） nodes/HP, and （1.93 ± 0. 13） respectively, all P 〈 0. 05].The A value, ALP staining positive rate, calcium node number, and type Ⅰ collagen expression score of the high glucose with drynaria total flavonoid group were （0. 352 ±0. 022）, 25. 3%, （4.50 ± 1.29）/HP, and （1.70 ± 0. 03） respectively, all significantly higher than those of the sodium glycerophophate ＋ vitamin C ＋dexamethasone [（0. 139 ±0.013）, 22.7%, （3.25 ± 1.50）/HP, and （1.28 ±0.27） respectively, all P 〈0. 05]. The AGE expression levels of the high glucose classical induction group and high glucose ＋drynaria total flavonoid group were both significantly higher than those of the low glucose classical induction group and low glucose ＋ ＋ drynaria total flavonoid group （both P 〈 0. 05） . Ther were no significant differences in the AGE level among the low glucose control, low glucose classical induction, and low glucose ＋drynaria total flavono

关 键 词：骨质疏松 细胞培养 成骨细胞 骨碎补总黄酮 

分 类 号：R686[医药卫生—骨科学]