Objective： To observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids （EFs） regulate the balance between osteogenic differentiation and adipogenic differentiation in b...Objective： To observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids （EFs） regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal ceils of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis. Methods： Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- （1 μg/mL）, medium- （10 μg/mL） and high- （100 μg/mL） dose EFs were administrated correspondingly. Alkaline phosphatase （ALP） staining, ALP activity determination, oil red O staining and real- time polymerese chain reaction （RT-PCR） were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. Moreover, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 （DKK1） was used in the medium group. Enzyme linked immunosorbent assay （ELISA） and RT-PCR were used to determine mRNA levels of 13-catenin, low density lipoprotein receptor-related protein 5 （LRP5） and T cell factor （TCF） protein, known as wnt/β-catenin signal pathway related factors. Results： EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 （Runx2）, osteocaicin and collagen I, and decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 （PPAR r/-2） and CCAAT enhancer-binding protein-α （C/EBP α） in a dose-dependent manner. While osteobiast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Also EFs up-regulated mRNA expression levels of β-catenin, LRP5 and展开更多
目的探讨续断皂苷VI调控Wnt/β-Catenin通路对骨质疏松的影响并分析其作用机制。方法构建SPF级Wistar大鼠骨质疏松症模型与健康大鼠共同饲养形成对照组,将50只雌性大鼠随机分为4组。依次设置为:续断皂苷VI治疗组,模型对照组,假手术组,...目的探讨续断皂苷VI调控Wnt/β-Catenin通路对骨质疏松的影响并分析其作用机制。方法构建SPF级Wistar大鼠骨质疏松症模型与健康大鼠共同饲养形成对照组,将50只雌性大鼠随机分为4组。依次设置为:续断皂苷VI治疗组,模型对照组,假手术组,空白对照组。使用不同双侧卵巢摘除手术法处理实验组大鼠。除空白对照组外,模型对照组与假手术组以及续断皂苷VI处理组的大鼠均制成骨质疏松症大鼠模型,将续断皂苷VI组的大鼠连续给药12周。使用X线骨密度仪骨密度(BMD,bone mineral density)测定法检测各组大鼠BMD,观察给药前后大鼠BMD变化情况;使用生物力学测定仪测定各组大鼠后肢股骨的最大载荷差异;使用RT-PCR实验和Western Blotting实验以检测不同组Wnt3a、β-catenin、LRP5、Runx2、SOST和Osx转录组与蛋白组表达各自发生的差异。结果续断皂苷VI治疗12周可显著提高骨质疏松症大鼠模型股骨的BMD,并提高椎体在最大负荷和弹性模量下的生物力学能力,降低了骨质疏松症在骨组织中造成的致病性。对于组织形态学方面的考察发现,续断皂苷VI治疗组在治疗持续12周后,小梁排列整齐,小梁稍变薄,股骨没有明显的轻微骨折。在进行续断皂苷VI治疗处理的情况下,经典Wnt/β-catenin信号通路中涉及的LRP5、β-catenin、Runx2和Osx的表达显著上调,而在该通路中SOST的表达下调(P<0.05)。结论续断皂苷VI可显著提高骨质疏松模型大鼠的骨密度,其保护机制与激活Wnt/β-catenin信号通路,上调骨质疏松大鼠骨中β-catenin、LRP5的mRNA表达有相关。展开更多
文摘Objective： To observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids （EFs） regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal ceils of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis. Methods： Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- （1 μg/mL）, medium- （10 μg/mL） and high- （100 μg/mL） dose EFs were administrated correspondingly. Alkaline phosphatase （ALP） staining, ALP activity determination, oil red O staining and real- time polymerese chain reaction （RT-PCR） were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. Moreover, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 （DKK1） was used in the medium group. Enzyme linked immunosorbent assay （ELISA） and RT-PCR were used to determine mRNA levels of 13-catenin, low density lipoprotein receptor-related protein 5 （LRP5） and T cell factor （TCF） protein, known as wnt/β-catenin signal pathway related factors. Results： EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 （Runx2）, osteocaicin and collagen I, and decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 （PPAR r/-2） and CCAAT enhancer-binding protein-α （C/EBP α） in a dose-dependent manner. While osteobiast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Also EFs up-regulated mRNA expression levels of β-catenin, LRP5 and
文摘目的探讨续断皂苷VI调控Wnt/β-Catenin通路对骨质疏松的影响并分析其作用机制。方法构建SPF级Wistar大鼠骨质疏松症模型与健康大鼠共同饲养形成对照组,将50只雌性大鼠随机分为4组。依次设置为:续断皂苷VI治疗组,模型对照组,假手术组,空白对照组。使用不同双侧卵巢摘除手术法处理实验组大鼠。除空白对照组外,模型对照组与假手术组以及续断皂苷VI处理组的大鼠均制成骨质疏松症大鼠模型,将续断皂苷VI组的大鼠连续给药12周。使用X线骨密度仪骨密度(BMD,bone mineral density)测定法检测各组大鼠BMD,观察给药前后大鼠BMD变化情况;使用生物力学测定仪测定各组大鼠后肢股骨的最大载荷差异;使用RT-PCR实验和Western Blotting实验以检测不同组Wnt3a、β-catenin、LRP5、Runx2、SOST和Osx转录组与蛋白组表达各自发生的差异。结果续断皂苷VI治疗12周可显著提高骨质疏松症大鼠模型股骨的BMD,并提高椎体在最大负荷和弹性模量下的生物力学能力,降低了骨质疏松症在骨组织中造成的致病性。对于组织形态学方面的考察发现,续断皂苷VI治疗组在治疗持续12周后,小梁排列整齐,小梁稍变薄,股骨没有明显的轻微骨折。在进行续断皂苷VI治疗处理的情况下,经典Wnt/β-catenin信号通路中涉及的LRP5、β-catenin、Runx2和Osx的表达显著上调,而在该通路中SOST的表达下调(P<0.05)。结论续断皂苷VI可显著提高骨质疏松模型大鼠的骨密度,其保护机制与激活Wnt/β-catenin信号通路,上调骨质疏松大鼠骨中β-catenin、LRP5的mRNA表达有相关。
