CAJ | 학술논문

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.
목적 료해골쇄보총황동재불동당농도하대골수간충질간세포(BMSC)성골분화적영향,위재당뇨병성골질소송증적방치제공이론의거.방법 대SD대서BMSC재체외조건하배양,진행정향성골유도분화,채용전골수첩벽법분리순화BMSC.실험분위저당조、고당조、저당유도조、고당유도조、저당골쇄보조、고당골쇄보조6조,검측상관성골지표감성린산매(ALP)활성、광화결절수목、효원단백.관찰골쇄보총황동재불동당농도하대BMSC성골분화적영향.결과 골쇄보총황동능촉진BMSC성골지표적표체,저당골쇄보조성골지표적표체고우저당유도조[ALP활성(0.439±0.024비0.385±0.029)、ALP염색양성솔(48.7%비35.0%)、광화결절수(9.75±1.71개/HP비6.25 ±0.96 개/HP)、Ⅰ형효원단백적분(2.21±0.07비1.93±0.13)]차이균유통계학의의(균P＜0.05).고당골쇄보조고우고당유도조[ALP활성(0.352±0.022비0.139±0.013)、ALP염색양성솔(25.3%비22.7%)、광화결절수(4.50±1.29개/HP비3.25±1.50개/HP)、Ⅰ형효원적분(1.70±0.03비1.28±0.27)]차이균유통계학의의(균P＜0.05).불동당농도조적당기화종말산물(AGE)수평수당농도화배양시간연장정증고추세(균P＜0.05),여Ⅰ형효원단백적표체정부상관(r=-0.410,P＜0.05).결론 골쇄보총황동가이촉진BMSC성골분화병감경고당조건하대성골분화적억제작용.
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 flavonoid groups (all P ＜ 0. 05); and among the high glucose control, high glucose classical induction, and high glucose + drynaria total flavonoid groups (all P ＞＜ 0. 05). However, the AGE levels of the high glicose groups were all significantly higher than those of the corresponding low glucose groups (all P ＜ 0. 05). Glucose increased the AGE levels dose- and time-dependently. The concentrations of AGEs were significantly negatively correlated with the expression of type Ⅰ collagen (r= - 0. 410, P ＜ 0. 05).Conclusions Drynaria total flavonoid promotes the osteogenic differentiation of BMSCs and relieves the inhibitory effect of osteogenic differentiation by glucose at high concentration. Thus drynaria total flavonoid may provide a potential therapy for diabetic osteoporosis.