散血草萃出物抑制破骨細胞的生成及骨再吸收

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Title:	散血草萃出物抑制破骨細胞的生成及骨再吸收 Suppressive effect of Ajuga bracteosa extracts on osteoclast formation invitro and resportion in vivo
Authors:	張惠詒;Hui-yi chang
Contributors:	醫學院基礎醫學研究所
Keywords:	去卵巢;鈣吸收;骨質疏鬆;散血草;osteoclast;bone resorption;Ajuga bracteosa
Date:	2010
Issue Date:	2010-09-29 12:13:25 (UTC+8)
Abstract:	散血草為台灣常用民間藥之一。本研究探討散血草酒精萃取物(ABEE) 對去卵巢大鼠所引起的骨質疏鬆症狀的改善效果，並以細胞實驗解明其作用機理。大鼠分為兩大組別，偽手術組與卵巢切除組，卵巢切除組再分成四組：卵巢切除經口給予ABEE (400、1200 mg/kg )、去離子水、alendronate (2.5 mg/kg，每個星期經口給藥三次)。大鼠經卵巢切除後4個星期開始給藥，持續給藥12週。ABEE和alendronate顯著降低c端telopeptides 第一型膠原蛋白的濃度及鹼性磷酸酶的活性。ABEE及alendronate能增加股骨的骨密度及骨鈣含量。Micro CT分析的結果顯示， ABEE及alendronate 處理組的股骨骨小樑體積 / 組織體積的比率和骨小樑數目高於卵巢切除組。組織學的分析也顯示ABEE和alendronate也會增加股骨彎曲能量。散血草所有分層皆能抑制核因子?羠受體活化因子配體 (receptor activator of nuclear factor-κB ligand；RANKL) 誘導破骨細胞的分化，尤以氯仿層的第四分層作用最明顯。RANKL可透過NF-κB及MAPKs路徑使骨髓幹細胞和小鼠巨噬細胞株RAW264.7分化成破骨細胞，而研究中發現ABCE4可透過抑制RANKL引起的NF-κB, AP-1表現，達到抑制破骨細胞分化的效果結論：口服ABEE能抑制卵巢切除引起的大鼠骨質疏鬆，其作用機轉可經由抑制破骨細胞的形成而降低卵巢大鼠的骨質流失。 Ajuga bracteosa is a famous folk medicine in Taiwan. This research was to investigate the ameliorative effect of crude ethanol extract of A.bracteosa (ABEE) on osteopenia in ovariectomized (OVX) rats and the mechanism through in vitro experiments. All of rats were divided into sham and OVX groups. The OVX rats were allowed to lose bone for 4 weeks. At 4th week post OVX operation, the OVX rats were dived into four groups treated with water, ABEE (400, 1200 mg/kg, daily, orally) and alendronate (2.5 mg/kg, p.o., 3 times per week) for 12 weeks. ABEE and alendronate decreased plasma levels of C-terminal telopeptides type I collagen and alkaline phosphatase. ABEE and alendronate increased the bone density and calcium content in the femur of OVX rats. The micro-CT analysis indicated the ratio of trabeculae bone volume to tissue volume and trabeculae number of ABEE and alendronate treated groups were higher than OVX group. The histomorphometric analysis also showed that ABEE and alendronate treatments suppressed the osteoclast surface per bone surface in the tibia of OVX rats. ABEE and alendronate could also increase the OVX-induced decrease in bending energy of femurs. It is found in this experiment that all of the A. bracteosa fractions could inhibit osteoclast differentiation induced by receptor activator nuclear factor kappa B (RANKL), with the fraction-IV in chloroform layer (ABCE4) being the most active. Receptor activator nuclear factor kappa B (RANKL) induced osteoclastogenesis through NF-κB and MAPKs pathways in bone marrow stromal cells and murine macrophage cell line RAW264.7 cells. This study was found ABCE4 could markedly suppressed NF-κB and AP-1 expressions which induced by RANKL to decrease osteoclastogenesis. In conclusion, oral administration of ABE could suppress osteoporosis in OVX rats, the mechanism may through inhibit osteoclastogenesis led to decrease bone mass loss in OVX rats.
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