Effect of puerarin on the P13K pathway for glucose transportation and insulin signal transduction in adipocytes

To explore the effect of puerarin on insulin receptor (IR), insulin receptor substrate-1 (IRS-1) and protein expression of protein kinase B(PKB) in the P13K pathway of the glucose consumption, transportation and insulin signal transduction in 3T3-L1 adipocytes with insulin resistance. The insulin resistance 3T3-L1 adipocytes model was established by free fatty acid induction. The model cells were managed with puerarin in different concentrations. Glucose consumption was detected with glucose oxidase method, glucose transportation rate was determined by 2-deoxy-3H glucose ingesting method, and the IR, IRS-1 and PKB expression were determined by Western blot. Glucose consumption and transportation were significantly decreased in the model adipocytes, but increased after treated with puerarin (P<0.01). Moreover, the level of tyrosine phosphorylation of IR subunit β was higher in the puerarin treated groups, and that of IRS-1 was higher in the group treated with low dose puerarin than that in the model group. The 3T3-L1 adipocytes of insulin resistance model could be induced by free fatty acid successfully, puerarin could promote the glucose utilization in them to alleviate the insulin resistance, which may be related with the action in advancing the tyrosine phosphorylation of IR and IRS-1.     

不同溶剂对野葛中葛根素提取效果影响的研究

目的考察不同溶剂对葛根素提取效果的影响。方法以水和不同浓度的乙醇提取野葛根中的葛根素,以葛根素的提取率和含量为考察指标,通过L9(34)正交试验,比较不同溶剂提取的最佳条件。结果热水提取法的最适条件为料水比1∶10、提取时间2.0 h、提取次数2次;醇提的最佳条件为料液比1∶6,提取时间2.0 h,乙醇体积分数为30%,提取3次。结论优选的两种溶剂提取工艺条件都合理可行,相比之下,水提取的综合指标更高一些。     

超临界CO2溶胀过程中葛根素乙醇溶液的溶解特性研究

为了设计用超临界流体抗溶剂结晶(GAS)法制备葛根素的超细颗粒,用静态平衡法考察了超临界CO2溶胀过程中葛根素在乙醇溶液的溶解特性.实验结果表明,体系压力变化对溶解度的影响最为显著.同时,我们发现,伴随着体系压力的递增,溶液中葛根素浓度表现为先扬后抑的趋势,甚至大于其初始值.由此可知,GAS过程CO2兼有抗溶剂和萃取剂的双重功能:前者降低溶质的溶解度,后者则提高溶液浓度.     

葛根素的电喷雾电离裂解规律解析

利用电喷雾电离技术对葛根素的结构和质谱裂解规律进行研究,用Mass frontier3.0软件辅助解析了主要特征碎片离子及其裂解途径。结果发现,葛根素主要通过D环的糖苷碎裂而发生裂解,出现碎片m/z399、381、363、351、335、321。碎片m/z399裂解成碎片m/z381,碎片m/z381又裂解成碎片m/z363和351为ESI-MS裂解特征。     

HPLC测定山葛降脂滴丸中葛根素的含量

目的采用高效液相色谱法(HPLC)对山葛降脂滴丸中葛根素含量进行测定。方法色谱柱为Diaomonsil C18色谱柱(250×4.6 mm,5μm)、ODS预柱,以甲醇-0.2%磷酸溶液(28∶72),检测波长为250 nm。结果葛根素在11.75~188.0μg/mL内,浓度与相应峰面积呈良好的线性关系(r=0.9999),平均回收率为99.50%,RSD为1.44%。结论该方法对山葛降脂滴丸中葛根素含量的测定简便易行,结果准确可靠,适用于对山葛降脂滴丸进行质量控制。     

高效液相色谱法测定纳豆胶囊中的葛根素

建立了富含葛根素的纳豆胶囊中葛根素的高效液相色谱分析方法。样品经过体积分数70%的甲醇提取后,采用ZORBAX ODS C18（5 μm, 4.6 mm×200 mm)色谱柱分离,以甲醇-36%乙酸-水（25∶3∶72）为流动相进行洗脱,流速1.0 mL/min,紫外检测波长247 nm。结果表明,葛根素在0～1 000 mg/L范围内线性关系良好（R2=0.999 4）,检出限是0.005 mg/g。在10 mg/g、20 mg/g、30 mg/g添加水平下葛根素的平均回收率为99.5%～104.5%,相对标准偏差为3.8%。该方法简单快速、回收率高、准确性好,可用于富含葛根素的纳豆胶囊中葛根素含量分析。     

HPLC 法测定葛根超细粉中葛根素含量

建立一种测定葛根超微粉中葛根素含量的方法。采用RP-HPLC法测定葛根素的含量。色谱柱为Agilent SGE protecol C18（5μm 4．6×250mm）,流动相为甲醇-水（25∶75）,流速为1．0mL/min,柱温为35℃,检测波长为250nm。葛根素在47～470μg/mL的线性关系良好,R＝0．9999。该方法操作简便、稳定可靠,可用于葛根超微粉的葛根素含量测定。     

Improvement in solubility and bioavailability of puerarin by mechanochemical preparation

An efficient and solvent-free procedure was developed to improve the solubility and bioavailability of the poorly water-soluble drug puerarin by mechanochemical technology. The stable inclusion complex of puerarin and 2-hydroxypropyl-β-cyclodextrin (HPCD) was prepared by a ball mill under the following conditions: equimolar ratio of puerarin to HPCD; rotational speed of 250 rpm; milling time of 90 min; steel balls of 22 mm diameter. The solid complex was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), powder X-ray diffractometry (XRD), and fourier transformation-infrared spectroscopy (FT-IR). Mechanochemical action could result in enhanced molecular encapsulation, homogeneous distribution and amorphization of the drug. In comparison to puerarin, a 1.64-fold increase in absolute bioavailability was obtained. The solubility of the inclusion complex was 25.33-fold higher and the drug release amount reached 79.44% at 15 min, 2.76-fold higher.     

葛根素对21日龄樱桃谷鸭钙磷代谢的影响

选用健康的1日龄的樱桃谷鸭128只,在基础日粮预饲一周后,分别饲喂在基础日粮中添加不同水平葛根素的日粮,于21日龄屠宰对血清及骨骼里的钙、磷及相关指标进行检测.结果表明:低水平的葛根素(5 mg/kg)对骨的形成有利,可以有效阻止骨的吸收,促进骨的形成;高水平的葛根素对骨的形成不利,其主要作用机理是雌激素样作用.     

固体分散技术制备葛根素亲水凝胶缓释片

采用固体分散技术制备葛根素亲水凝胶缓释片。先采用单因素实验制备葛根素固体分散体,然后通过正交试验考察HPMC的K4M型和K15M型的质量比m(K4M):m(K15M)(A),HPMC的质量分数(B),淀粉与乳糖质量比m(淀粉):m(乳糖)(C),淀粉与乳糖合计的质量分数(D)4个因素对葛根素亲水凝胶缓释片的影响。得到葛根素固体分散体的最佳制备工艺:以PEG4000为载体,药物与载体的质量比为1:6;葛根素亲水凝胶缓释片的最佳制备工艺为:HPMC的K4M型和K15M型质量比m(K4M):m(K15M)=1:2,HPMC的质量分数为片重的30%,淀粉与乳糖的质量比m(淀粉):m(乳糖)=1:3,淀粉与乳糖合计的质量分数为片重的40%。该缓释片处方合理,工艺可行,达到缓释的预期目的。