黄芪和当归的主要活性成分配伍促进衰老造血干细胞增殖作用的研究

目的探讨黄芪和当归的主要活性成分黄芪甲苷、芒柄花素、毛蕊异黄酮、毛蕊异黄酮苷和阿魏酸配伍对衰老造血干细胞(HSCs)增殖的影响,并从细胞周期调控研究其作用机制。方法以三丁基过氧化氢(T-BHP)构建小鼠HSCs衰老模型,研究不同质量浓度5种活性成分对HSCs衰老和增殖的影响,寻找可促进细胞增殖的主要活性成分。在此基础上,采用正交试验研究5种活性成分配伍对HSCs衰老和增殖的影响,寻找5种活性成分促进细胞增殖的合理配伍。最后,采用HSCs衰老模型,制备黄芪-当归1∶1配伍含药血浆,设立对照组、模型组、空白血浆组、阿魏酸组、黄芪甲苷组、芒柄花素组、毛蕊异黄酮组、毛蕊异黄酮苷组、活性成分配伍组、黄芪-当归1∶1含药血浆组,药物作用于衰老细胞,以SA-β-半乳糖苷酶(SA-β-Gal)染色法测定HSCs衰老率,CCK-8法测定细胞增殖率,流式细胞术检测细胞周期分布,Western blotting法检测Cyclin D1、细胞周期蛋白依赖性激酶4(CDK4)蛋白表达。结果阿魏酸、黄芪甲苷、芒柄花素均可显著促进衰老HSCs增殖和降低细胞衰老率,而毛蕊异黄酮、毛蕊异黄酮苷单独应用对HSCs增殖和细胞衰老率无显著影响。正交试验发现,以阿魏酸、芒柄花素和黄芪甲苷为基础因子,毛蕊异黄酮和毛蕊异黄酮苷为次要因子进行配伍的5种活性成分组合促细胞增殖和降低细胞衰老率的作用最强。阿魏酸、黄芪甲苷、芒柄花素、活性成分配伍、含药血浆均可降低细胞衰老率,使G0/G1期细胞减少,G2/M+S期细胞增加,并使Cyclin D1和CDK4蛋白表达增强。以上效应以活性成分配伍组和含药血浆组最强。结论黄芪和当归的主要活性成分阿魏酸、黄芪甲苷和芒柄花素对衰老HSCs具有促增殖和改善衰老作用,而毛蕊异黄酮和毛蕊异黄酮苷单用时并无明显的作用。当5种活性成分配伍后对衰老HSCs的促增殖作用最强,并可改善HSCs衰老,促进衰老HSCs由静止期向增殖期转换。黄芪和当归的主要活性成分及其配伍促HSCs增殖和抗HSCs衰老的作用可能与调节细胞周期相关蛋白表达及促进细胞周期转换有关。     

红芪中5种黄酮类成分对大鼠骨髓间充质干细胞和成骨细胞成骨分化的影响

目的研究红芪中毛蕊异黄酮、芒柄花素、芒柄花苷、异甘草素和美迪紫檀素5种黄酮类成分对大鼠骨髓间充质干细胞(rat bone marrow stromal cells,rBMSCs)和颅骨成骨细胞(rat calvarial osteoblasts,ROBs)成骨性分化的影响。方法贴壁分离法培养rBMSCs;酶消解法分离培养ROBs;MTT法检测rBMSCs和ROBs细胞的增殖情况;试剂盒检测rBMSCs和ROBs的碱性磷酸酶(ALP)活性和Ca~(2+)含量;茜素红染色法检测矿化结节形成。结果 5种成分均能不同程度促进rBMSCs和ROBs的增殖,提高ALP活性,增加Ca~(2+)含量,增加钙化结节面积和数量(P0.05)。其中毛蕊异黄酮促进rBMSCs的成骨分化作用最佳,美迪紫檀素促进ROBs的成骨分化作用最佳,毛蕊异黄酮次之。结论这5种黄酮类成分对rBMSCs和ROBs的成骨功能均有一定程度地改善作用,而毛蕊异黄酮对2种细胞表现的成骨活性均较佳,可发展作为良好的骨诱导活性因子。     

芒柄花素对围绝经期女性的药用价值研究

针对女性围绝经期综合征，传统采用雌激素替代疗法(hormone replacement therapy，HRT)进行治疗，但长期采用HRT可增加发生阴道出血、乳腺癌及子宫内膜癌等疾病的风险。因此相对安全有效的植物雌激素越来越受到人们的关注。芒柄花素(formononetin，FMN)作为天然来源的异黄酮类植物雌激素，具有抗肿瘤、抗氧化、抗炎、调节免疫、雌激素样及调节肠道菌群等多种作用，能防治骨质疏松、糖尿病和心脑血管疾病，对于改善女性卵巢功能减退导致的围绝经期症状具有重要意义。本文针对FMN的药用价值对于改善围绝经期女性相关症状方面进行综述，为FMN的应用提供参考。     

复方参芪冲剂质量标准的研究

OBJECTIVE To establish the quality standards for Fufangshenqi Granules.METHODS Radix Astragali,Radix Sophorae Flavescentis and Radix Glycyrrhizae in Fufangshenqi Granules were identified with TLC;Total saponin in Fufangshenqi Granules was determined by sp     

川产道地药材梭果黄芪中毛蕊异黄酮葡萄糖苷和芒柄花素的含量测定

目的：建立梭果黄芪药材中毛蕊异黄酮葡萄糖苷和芒柄花素含量测定方法,为完善梭果黄芪的质量标准提供依据。方法：Agilent TC-C18（4.6 mm×250 mm,5μm）色谱柱;以乙腈为流动相A,以水（含0.2%甲酸）为流动相B,梯度洗脱;检测波长260 nm;柱温30℃;流速1.0 mL/min;进样量10μL。结果：毛蕊异黄酮葡萄糖苷和芒柄花素的线性回归方程分别为Y=30.025 X＋5.1096（r=0.999 7）和Y=58.24 X-94.279（r=0.999 7）;线性范围分别为5.52～110.40μg/mL和5.54～110.72μg/mL;样品中毛蕊异黄酮葡萄糖苷和芒柄花素的总量在0.0053%～0.0179%之间。结论：本实验建立的毛蕊异黄酮葡萄糖苷和芒柄花素的含量测定方法简便、准确,可用于梭果黄芪药材中黄酮类成分的质量控制。     

RP-HPLC波长切换法同时测定消栓通络片中4种成分的含量

目的建立同时测定消栓通络片中芦丁、丹酚酸B、肉桂酸和芒柄花素4种成分含量的RP-HPLC方法。方法采用Diamonsil C18柱(200 mm×4.6 mm,5μm),以乙腈-体积分数为0.1%磷酸水溶液进行梯度洗脱;流速:1.0 mL.min-1;柱温:30℃;采用切换波长的方法:在360 nm下检测芦丁;在288 nm下检测丹酚酸B和肉桂酸;在248 nm下检测芒柄花素。结果芦丁、丹酚酸B、肉桂酸和芒柄花素的质量浓度与峰面积分别在14.3～85.8 mg.L-1(r=0.999 4,n=6)、10.56～63.36mg.L-1(r=0.999 3,n=6)、1.8～10.8 mg.L-1(r=0.999 8,n=6)和0.58～3.48 mg.L-1(r=0.999 6,n=6)内呈良好的线性关系;平均加样回收率依次为98.4%、99.3%、99.2%和98.4%。RSD分别为1.4%、0.9%、1.3%和1.5%。结论该方法适合于同时测定消栓通络片中芦丁、丹酚酸B、肉桂酸和芒柄花素的含量。     

Formononetin,a novel FGFR2 inhibitor,potently inhibits angiogenesis and tumor growth in preclinical models

Most anti-angiogenic therapies currently being evaluated in clinical trials target vascular endothelial growth factor (VEGF) pathway, however, the tumor vasculature can acquire resistance to VEGF-targeted therapy by shifting to other angiogenesis mechanisms. Therefore, other potential therapeutic agents that block non-VEGF angiogenic pathways need to be evaluated. Here we identified formononetin as a novel agent with potential anti-angiogenic and anti-cancer activities. Formononetin demonstrated inhibition of endothelial cell proliferation, migration, and tube formation in response to basic fibroblast growth factor 2 (FGF2). In ex vivo and in vivo angiogenesis assays, formononetin suppressed FGF2-induced microvessel sprouting of rat aortic rings and angiogenesis. To understand the underlying molecular basis, we examined the effects of formononetin on different molecular components in treated endothelial cell, and found that formononetin suppressed FGF2-triggered activation of FGFR2 and protein kinase B (Akt) signaling. Moreover, formononetin directly inhibited proliferation and blocked the oncogenic signaling pathways in breast cancer cell. In vivo, using xenograft models of breast cancer, formononetin showed growth-inhibitory activity associated with inhibition of tumor angiogenesis. Moreover, formononetin enhanced the effect of VEGFR2 inhibitor sunitinib on tumor growth inhibition. Taken together, our results indicate that formononetin targets the FGFR2-mediated Akt signaling pathway, leading to the suppression of tumor growth and angiogenesis.     

Vasorelaxant effect of formononetin in the rat thoracic aorta and its mechanisms

The purpose of the present study was to investigate the effect of formononetin and the related mechanisms on isolated rat thoracic aorta. Formononetin concentration dependently relaxed aortic rings precontracted with norepinephrine (NE, 1 μM) or KCl (80 mM). Pretreatment with formononetin noncompetitively inhibited contractile responses of aortas to NE and KCl. The vasorelaxant effect of formononetin partially relied on intact endothelia, which was significantly attenuated by incubation with N^ω-nitro-l-arginine methyl ester (100 μM). In endothelium-denuded rings, glibenclamide (10 μM) and tetraethylammonium (5 mM) showed slight reduction in the vasorelaxant effect of formononetin. Moreover, formononetin reduced NE-induced transient contraction in Ca²⁺-free solution and inhibited the vasocontraction induced by increasing external calcium in medium plus 80 mM KCl. Our results suggested that formononetin induced relaxation in rat aortic rings through an endothelium-dependent manner via nitric oxide synthesis pathway, and also involving an endothelium-independent vasodilatation by the blockade of Ca²⁺ channels. The opening of K⁺ channels might also be one of the mechanisms of formononetin-induced vasorelaxation.     

二次通用旋转组合法优化黄芪中毛蕊异黄酮苷和芒柄花素的酶解提取工艺

目的采用二次通用旋转组合设计优化黄芪中毛蕊异黄酮苷和芒柄花素的酶解提取工艺。方法在确定酶比例的基础上,选定复合酶用量、酶解时间、加水量和提取时间为考察因素,以毛蕊异黄酮苷、芒柄花素提取量为指标,采用二次通用旋转组合设计优化黄芪酶解提取工艺。结果优化所得黄芪酶解提取的最佳工艺条件:复合酶用量340 mg,酶解时间110min,加水量19倍,提取时间150 min,在此条件下,毛蕊异黄酮苷、芒柄花素提取量分别为0.25 mg/g、67.95μg/g。结论优化所得黄芪酶解提取工艺稳定可行。