葛根对地塞米松诱导的胰岛素抵抗的影响

目的 :探讨葛根提取物对地塞米松造成的胰岛素抵抗的影响及其作用机理。方法 :采用地塞米松诱导 3T3-L1脂肪细胞胰岛素抵抗 ,检测药物对细胞培养基中葡萄糖浓度的影响 ;采用肌肉注射地塞米松 ( 1mg·kg^-1,隔日 1次 )的方法建立胰岛素抵抗动物模型 ,测定空腹血糖 (FBG)、血清胰岛素 (FINS) ,并计算胰岛素抵抗指数 (IRI)与胰岛素敏感指数 (ISI)。结果 :葛根提取物能够明显降低胰岛素抵抗 3T3-L1脂肪细胞培养基中的葡萄糖水平 ,增强细胞对胰岛素的敏感性 ;葛根提取物显著降低胰岛素抵抗大鼠空腹血清胰岛素水平及胰岛素抵抗指数 ,有效改善了大鼠的胰岛素抵抗状态。结论 :葛根对地塞米松造成的胰岛素抵抗具有明显的改善作用。     

A herbal formula containing roots of Salvia miltiorrhiza (Danshen) and Pueraria lobata (Gegen) inhibits inflammatory mediators in LPS-stimulated RAW 264.7 macrophages through inhibition of nuclear factor κB (NFκB) pathway

Ethnopharmacological relevance

The herbal formula DG, containing roots of Salvia miltiorrhiza (Danshen) and Pueraria lobata (Gegen), has long history in treating cardiovascular diseases. It has been shown to be able to reduce intima-media thickening in coronary patients in our previous clinical study. Since intima-media thickening is the hallmark of atherosclerotic disease, the etiology of which is inflammation of the arterial wall, the mechanism underlying the effect of DG may be related to its anti-inflammatory activities.

Aim of study

The present study aims to determine the anti-inflammatory activity of DG and elucidate its underlying mechanisms with regards to its molecular basis of action.

Materials and method

The anti-inflammatory effect of DG was studied by using lipopolysaccharide (LPS)-stimulated activation of nuclear factor κB (NFκB) pathway and subsequent production of inflammatory mediators, including nitric oxide (NO), prostaglandin E₂ (PGE₂), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and macrophage chemotactic protein-1 (MCP-1), in mouse RAW 264.7 macrophages.

Results

The present study demonstrated that DG could suppress the production of NO and PGE₂ through the inhibition of iNOS and COX-2 genes. DG could also inhibit the production of IL-1β, IL-6 and MCP-1, but not TNF-α, through the inhibition of respective mRNA expressions. Further investigations showed the inhibitory effect of DG on activation of IKKα/β and degradation of IκBα, thus preventing nuclear translocation of NFκB. All these results suggested the inhibitory effects of DG on the production of inflammatory mediators through the inhibition of the NFκB pathway.

Conclusions

The inhibitory effects of DG on the production of inflammatory mediators by LPS-stimulated RAW 264.7 macrophages, are accomplished by inhibiting the nuclear translocation of NFκB through inactivating IKKα/β and preventing degradation of IκBα.     

葛根总黄酮磷脂复合物制备工艺的研究

目的：优选葛根总黄酮磷脂复合物最佳成型工艺。方法：以磷脂复合物的复合率和表观油水分配系数为指标,采用单因素正交试验优化葛根总黄酮磷脂复合物处方。结果：优选出制备工艺的最佳条件：葛根总黄酮和大豆卵磷脂质量比为1:4,反应溶剂为甲醇,葛根总黄酮的浓度为4mg·mL^-1,反应温度为55℃,反应时间为2h。按得到的优化条件制备的磷脂复合物的复合率为102.98%;pH=3时,Papp=0.392;pH=6.8时,Papp=0.323;综合得分为95.11分。结论：优化所得葛根总黄酮磷脂复合物处方合理,复合率高,油水分配系数增高,脂溶性得到改善。     

基于FTIR 和HPLC 的葛根化学成分整体分析与评价

目的：对不同产地葛根与粉葛的化学成分进行整体分析。方法：采用FTIR 和HPLC 分析方法,对FTIR 分析结果进行二阶导数谱、共有峰率和变异峰率的双指标序列分析和聚类分析;以葛根素为参比峰,对各产地葛根进行HPLC 指纹图谱相似度及聚类分析。结果：葛根在3 400~3 340、2 930、1 730、1 630、1 520、1 450、1 260、1 160、1 050~1 000、930、860~840、765、580 cm-1 等处有明显的吸收峰,主要为葛根素与淀粉的吸收峰。粉葛在上述吸收峰处也存在明显的吸收,说明其两者化学成分基本相似,但在红外指纹区有明显的区别,其中粉葛在1 460、1 420 和720 cm-1 处有特征裂解峰存在。葛根与其HPLC共有峰的相似度在81.4%~99.3%之间。结论：从葛根素的药用效果上考虑要优先使用葛根,FTIR 的方法简便易行且能为药材提供丰富的信息。     

Purification of Puerarin from Pueraria lobata by FCPC versus HSCCC Using Small-volume Columns

Objective To develop an efficient method for separating and purifying puerarin from the roots of Pueraria lobata.Methods Separation by fast centrifugal partition chromatography(FCPC)was processed with a biphasic solvent system composed of ethyl acetate-n-butanol-water(2:1:3).The separation conditions were determined as follows:sample loading of 10 mg,flow rate of 2 mL/min,rotation speed of 2200 r/min,ascending mode,and detection wavelength of 254 nm.High speed countercurrent chromatography(HSCCC)was used as a comparative method with the rotation speed of 800 r/min,flow rate of 2.0 mL/min,and sample loading of 10 mg in tail-to-head mode.Results Puerarin was obtained by FCPC with a resolution of 0.90 and a purity above 99%,while a resolution below 0.50 and a purity below 90%by HSCCC.Compared with HSCCC,FCPC has the advantages with higher purity and better resolution.Conclusion FCPC is a powerful method to separate and purify puerarin.     

RP-HPLC测定大鼠口服葛根后血浆中大豆苷元的含量

目的:建立准确快速测定大鼠口服葛根后血浆中大豆苷元含量的方法,探讨大豆苷元的体内代谢情况.方法:采用反相高效液相色谱法(RP-HPLC)测定大鼠口服药物后不同时间血浆中大豆苷元的含量,待测血浆样品采用沉淀蛋白法处理,Phenomenex Luna C18高效液相色谱柱(4.60 mm×250 mm,5μm)进行分析,流动相为甲醇-0.1%甲酸水(60∶40v/v).结果:测定方法稳定,给药后5 min血浆中即可检测出大豆苷元,20min内达到峰值,高、中、中低、低4种浓度的日内精密度均≤5.7%,日19精密度均≤9.3%,回收率分别为89.5%、93.2%、93.9%、84.8%,在0.1～64μg/mL范围内线性关系良好.结论:此方法快捷、灵敏、简便,用于血浆中大豆苷元的含量测定,取得满意结果.     

Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison with Pueraria lobata

Pueraria mirifica tubers collected from 28 out of 76 provinces of Thailand and Pueraria lobata tubers collected from Guangzhou province, China were submitted to HPLC analysis with the established gradient system comprising 1.5% acetic acid and acetonitrile. Five major isoflavonoids, including puerarin, daidzin, genistin, daidzein and genistein, were adopted as authentic standards. P. mirifica tubers showed intra- as well as inter-provincial differences in isoflavonoid and total isoflavonoid contents. The difference in both cases should be mostly influenced by genetic and environmental factors. In comparison with P. lobata, P. mirifica population exhibited differences only with a lower amount of daidzein.