丹参水提物和丹参素促进成骨细胞活性和防治泼尼松所致大鼠骨质疏松

目的　研究丹参水提物 (DWE)对糖皮质激素造成大鼠骨质疏松的预防作用 ,同时研究DWE和有效成分丹参素对体外培养的大鼠成骨细胞的影响 ,以探讨其作用机制。方法　① 4mon龄SD大鼠分为对照组、模型组 (以泼尼松 2 7mg·kg- 1·d- 1灌胃 ) ,丹参水提物组 (泼尼松 +DWE 5 0g·kg- 1·d- 1)和综合治疗组 (泼尼松 +司坦唑醇 0 5mg·kg- 1·d- 1+VitD3 2 50IU·kg- 1·d- 1+葡萄糖酸钙 0 5g·kg- 1·d- 1) ,共给药 12wk。采用骨组织形态计量学方法定量观察大鼠胫骨松质骨的骨结构 ,同时测定骨无机及有机质含量 ;②采用体外大鼠成骨细胞培养 ,测定DWE和丹参素对细胞碱性磷酸酶 (ALP)的活性及药物量效、时效关系。结果　泼尼松可导致骨小梁面积明显减少 ,骨结构异常 ,破骨细胞增多和骨形成率下降 ,伴随骨无机质钙盐 (Ca)和有机质羟脯胺酸 (HyP)下降 ,血钙上升 (以上指标P <0 0 1)。DWE完全对抗由泼尼松引起的上述异常并增加骨干重 ,对增加骨干重和骨有机质含量优于综合组。DWE和丹参素均可促进大鼠颅骨成骨细胞ALP活性 ,呈量效关系 (DWE最佳作用浓度为 1 0～ 2 0g·L- 1,丹参素为 0 1～ 1 0mg·L- 1)和时效关系 (随时间延长作用明显增强 ) ,DWE的效应按含丹参素量计算 ,与丹参素作用相当。结论　丹参水提?

Preventing cancellous bone loss in steroid-treated rats and stimulating bone formation by water extract of salvia miltiorrhixa and danshensu

AIMTo investigate the effects of water extract fractio n from Salvia Miltiorrhixa Bunge (DWE) and its bioactive compound danshensu on p rednisone-induced osteoporosis in rats and to examine the effect of DWE and dan shensu on activity of ALP in rat primary cultured osteoblasts. METHODSStudy 1: Thirty-two 4-month-old SD rats were randomly divided into four groups: ①intact rats was given with vehicle as control (Intact + Veh); ②Predn isone was given orally to the rats at dose of 2 7 mg·kg -1 ·d -1 as the model group (Pred+Veh); ③ DWE at dose of 5 0 g·kg -1 ·d -1 were added to the prednison treated rats (Pred+DWE) ; ④Conbined medicine (Stanozolo l 0 5 mg·kg -1 ·d -1 +VitD 3 25 IU·kg -1 ·d -1 +Calcium 0 5 g·kg -1 ·d -1 ) were added to the prednison treated rats (Pred+SD C). Double in vivo fluorochrome labeling was performed before killing the rats. The undecalcified longitudinal proximal tibial metaphyseal section were used to do bone histomorphometric analysis. The right ulna was used to test the bone cal cium content and bone hydroxyproline content. Study 2: New born SD rat calvarium osteoblasts were cultured in DMEM alone or exposed to different concentration o f DWE or danshensu for 2 and 10 days respectively, the activity of alkaline phos phatase (ALP) was detected with enzyme-chemistry method. RESULTSCompared with control group, trabecular bone volume and bone formation rate/TV in prednisone treated group dec reased by 41 3% (P<0 01) and 39 9% (P<0 01) respectively, osteoclast surface increased significantly accompanied with decrease of bone calcium and h ydroxyproline contents. The prednison plus DWE group was completely prevented fr om bone loss and showed increased bone formation rate and decreased osteoclast s urface when compared with prednison alone. The effects of DWE on the bone dried weight and bone organic content (hydroxyproline) were better than that of combi ned medicine treated group. DWE (the concentration from 0 5 to 10 0 g ·L -1 ) and danshensu (the concentration from 0 1 to 5 0 mg·L -1 ) increased ALP activity of rat cultured osteoblasts by two folds in a dose and time dependent manner. CONCLUSONDWE can protect trabecu lar bone loss from steroid-induced osteoporosis due to stimulating osteoblast a ctivity and decreasing osteoclast activity.