接骨木叶总黄酮抗骨质疏松作用研究
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摘要
目的:优化接骨木叶中总黄酮的提取工艺,建立骨质疏松模型,研究接骨木叶总黄酮的抗骨质疏松作用。方法:以接骨木叶为原料,以总黄酮得率为响应值,分别考察乙醇体积分数、液料比、温度等几个因素对接骨木叶总黄酮得率的影响,在单因素实验的基础上采用响应曲面法中的Box-Behnken设计,优化接骨木叶总黄酮提取工艺。建立链脲佐菌素小鼠骨质疏松模型,检测小鼠血清中ALP、ACP、Ca、P、TRAP、BGP等指标变化以及HE染色观察骨组织形态。结果:接骨木叶总黄酮的最佳提取工艺条件为提取温度70℃、提取时间60 min、乙醇体积分数70%、液料比70:1(mL:g),平均得率为7.249%,其所得总黄酮纯度为31.3%。抗骨质疏松作用研究中,接骨木叶总黄酮各剂量组中钙、磷、ALP、ACP、TRAP骨钙素水平差异显著,并随着接骨木叶总黄酮浓度的增加,血清钙水平、ALP水平显著降低(P<0.05,P<0.01),ACP、磷、TRAP水平显著升高(P<0.05,P<0.01)。从病理切片来看,随着接骨木叶总黄酮剂量的增加,骨基质明显增多,骨小梁增厚,成骨细胞增多,破骨细胞减少,骨密度增加。结论:接骨木叶总黄酮对链脲佐菌素小鼠骨质疏松模型效果显著,能起到良好的抗骨质疏松作用。
Objective: To optimize the extraction process of total flavonoids from elderberry leaves and establish a model of osteoporosis to study the anti-osteoporosis effect of total flavonoids from elderberry leaves. Methods: Using elderberry leave as raw materials, the total flavonoids yield was taken as the response value. The effects of several factors such as ethanol volume fraction, liquid-to-liquid ratio and temperature on the yield of total flavonoids of elderberry leaves were studied. The Box-Behnken design in response surface methodology was used to optimize the extraction process of total flavonoids from elderberry leaves. The osteoporosis model of streptozotocin was established. The changes of ALP,ACP, Ca, P, TRAP and BGP in serum were detected and the morphology of bone was observed by HE staining. Results: The optimum extraction conditions for total flavonoids from elderberry leaves were extraction temperature 70 ℃, extraction time 60 min, ethanol volume fraction 70%, liquid-to-liquid ratio70:1(mL:g), and the average yield was 7.249%. The total flavonoid purity was 31.3%. In the study of anti-osteoporosis, the levels of calcium, phosphorus, ALP, ACP and TRAP osteocalcin in the different doses of total flavonoids from elderberry leaves were significantly different with the increase of total flavonoids from elderberry leaves, serum calcium levels and ALP levels were significantly decreased(P<0.05, P<0.01), ACP, phosphorus, TRAP levels were significantly increased(P<0.05, P<0.01). From the pathological section, with the increase of the total flavonoids dose, the bone matrix increased significantly,the trabecular bone thickened, the osteoblasts increased, the osteoclasts decreased, and the bone density increased. Conclusion: The total flavonoids from elderberry leaves have a significant effect on the osteoporosis model of streptozotocin mice, and can play a good role in anti-osteoporosis.
Objective: To optimize the extraction process of total flavonoids from elderberry leaves and establish a model of osteoporosis to study the anti-osteoporosis effect of total flavonoids from elderberry leaves. Methods: Using elderberry leave as raw materials, the total flavonoids yield was taken as the response value. The effects of several factors such as ethanol volume fraction, liquid-to-liquid ratio and temperature on the yield of total flavonoids of elderberry leaves were studied. The Box-Behnken design in response surface methodology was used to optimize the extraction process of total flavonoids from elderberry leaves. The osteoporosis model of streptozotocin was established. The changes of ALP,ACP, Ca, P, TRAP and BGP in serum were detected and the morphology of bone was observed by HE staining. Results: The optimum extraction conditions for total flavonoids from elderberry leaves were extraction temperature 70 ℃, extraction time 60 min, ethanol volume fraction 70%, liquid-to-liquid ratio70:1(mL:g), and the average yield was 7.249%. The total flavonoid purity was 31.3%. In the study of anti-osteoporosis, the levels of calcium, phosphorus, ALP, ACP and TRAP osteocalcin in the different doses of total flavonoids from elderberry leaves were significantly different with the increase of total flavonoids from elderberry leaves, serum calcium levels and ALP levels were significantly decreased(P<0.05, P<0.01), ACP, phosphorus, TRAP levels were significantly increased(P<0.05, P<0.01). From the pathological section, with the increase of the total flavonoids dose, the bone matrix increased significantly,the trabecular bone thickened, the osteoblasts increased, the osteoclasts decreased, and the bone density increased. Conclusion: The total flavonoids from elderberry leaves have a significant effect on the osteoporosis model of streptozotocin mice, and can play a good role in anti-osteoporosis.
引文
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[14]苏艳杰,陈亚辉,崔燎.植物黄酮抗骨质疏松作用研究进展[J].中国骨质疏松杂志,2014,20(5):562-566.
[15]肖辉辉.接骨木抗骨质疏松活性成分的深入研究[D].沈阳:沈阳药科大学,2011.
[2]王冰.绝经过渡期妇女血清FSH水平与CTx、TP1NP的相关性分析[D].杭州:浙江大学,2015.
[3]Sosa Henríquez M J.Gómez de Tejada Romero.Osteoporosis[J].Medicine-Programa de Formación Médica Continuada AcreditadoJune,2018:3499-3505.
[4]Kathryn Schultz BA,Jennifer Moriatis,Wolf MD.Emerging Technologies in Osteoporosis Diagnosis[J].The Journal of Hand Surgery,2018,(1).
[5]Sueli Rodrigues.Edy Sousade Brito,Ebenezerde Oliveira Silva.Elderberry-Sambucus nigra L[J].Exotic Fruits,2018:181-185.
[6]OuYang Fu,Liu Yuan,LI Rong.Five Lignans and an Iridoid from Sambucus williamsii[J].Chinese Journal of Natural Medicines,2011,9(1).
[7]胡彦武,关颖丽,张立威,等.高效液相色谱法测定接骨木茎叶果实中芦丁的含量[J].医药导报,2014,33(9):1222-1224.
[8]杨序娟.接骨木抗骨质疏松活性成分的研究[D].沈阳:沈阳药科大学,2005.
[9]苏新芳.接骨木叶片总黄酮提取纯化及抗氧化活性研究[D].临沂:山西师范大学,2016.
[10]张爽,刘淑英,于莉莉,等.响应面法优化东北接骨木总黄酮的超声波提取工艺[J].北方园艺,2012,(14):11-14.
[11]彭璐,赵云,许旭东,等.玛咖对维甲酸诱导的骨质疏松症的抑制作用[J].中药药理与临床,2014,30(3):98-100.
[12]刘帼静,呼晓雷,何发忠.2型糖尿病骨质疏松研究进展[J].中国药理学通报,2016,32(10):1333-1336.
[13]程敏,胡选生,程楠,等.墨旱莲石油醚提取物对STZ糖尿病大鼠生化指标及肾组织病理学的影响[J].中国药理学通报,2018,34(3):407-411.
[14]苏艳杰,陈亚辉,崔燎.植物黄酮抗骨质疏松作用研究进展[J].中国骨质疏松杂志,2014,20(5):562-566.
[15]肖辉辉.接骨木抗骨质疏松活性成分的深入研究[D].沈阳:沈阳药科大学,2011.
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