淫羊藿苷分离方法的研究

以淫羊藿甲醇粗提物为原料,以研究规模化分离提纯淫羊藿苷的方法为目的,研究了4种分离方法的分离效果和可行性。结果表明最佳的分离方法为:用30%乙醇对淫羊藿甲醇粗提取物进行超声波辅助提取两次,提取液直接流经聚酰胺柱,并用相同溶剂洗柱,浓缩洗脱液至小体积后依次用氯仿和乙酸乙酯分别萃取,乙酸乙酯萃取物中淫羊藿苷的含量为60%,淫羊藿苷的总收率为74.11%。经60%乙醇重结晶一次,得90.94%含量的淫羊藿苷,总收率为36.30%。该方法工艺简单,生产成本低,产品收率高,纯度好,是工业化生产淫羊藿苷的理想方法。     

淫羊霍苷抗胃癌转移机制的研究

目的:探讨淫羊霍苷抑制胃癌细胞系AGS粘附和移动的作用及机制。方法:不同浓度淫羊霍苷作用AGS细胞24h后,粘附实验检测细胞粘附率,划痕损伤实验检测迁移速度。结果:2、4、8、12、16μg/ml淫羊霍苷处理24h后,AGS细胞粘附率分别为73.65%、46.24%、43.28%、41.59%和20.36%,总体呈下降趋势,4μg/ml组与2μg/ml和16μg/ml组比较差异有显著统计学意义(P<0.01),与8μg/ml和12μg/ml组比较差异无统计学意义(P>0.05)。4μg/ml淫羊霍苷处理24h后AGS细胞迁移速度明显降低(P<0.01)。结论:淫羊霍苷通过抑制AGS细胞粘附和运动来发挥对肿瘤转移的抑制作用。     

Icariin attenuates high glucose-induced insulin resistance in C2C12 myotubes

AIM: To observe the potential effects of icariin on high glucose-induced insulin resistance in C2C12 myotubes and to investigate its underlying mechanisms. METHODS: The insulin resistance model was induced by high glucose (25 mmol/L) in the C2C12 myotubes. The effects of icariin on Akt phosphorylation at T308, glucose transporter 4 (GLUT4) membrane translocation, and glucose uptake were investigated in high glucose-treated C2C12 myotubes. The protein levels of phosphorylated proteins were determined by Western blot. The glucose uptake was measured by colorimetric method. The small interfering RNA (siRNA) was used to knockdown the expression of p38 MAPK. RESULTS: Icariin significantly increased insulin-stimulated Akt T308 phosphorylation in C2C12 myotubes treated with high glucose. Treatment with icariin at 25, 50 and 75 μmol/L for 24 h increased Akt T308 phosphorylation in a dose-dependent manner (P<0.05 or P<0.01). Treatment with icariin at 50 μmol/L for 12, 24 and 36 h increased Akt T308 phosphorylation in a time-dependent manner (P<0.05 or P<0.01). In addition, treatment with icariin at 50 μmol/L for 24 h significantly enhanced the expression of GLUT4 on plasma membrane (P<0.01) and 2-deoxyglucose (2-DG) uptake (P<0.01). Treatment with icariin recovered high glucose-reduced p38 MAPK phosphorylation (P<0.01). Pharmacological or genetic inhibition of p38 MAPK abolished the protective impacts of icariin on insulin-stimulated Akt T308 phosphorylation (P<0.01), GLUT4 plasma membrane translocation (P<0.01), and 2-DG uptake under high glucose condition (P<0.05). CONCLUSION: Icariin attenuates high glucose-induced insulin resistance in C2C12 myotubes by activating p38 MAPK.     

类黄酮异戊烯转移酶基因在酿酒酵母中的表达

[目的]对类黄酮异戊烯转移酶基因在酿酒酵母中的表达进行研究。[方法]以粗毛淫羊藿为材料,对其类黄酮异戊烯转移酶基因(EaPT1)进行克隆,并构建重组质粒,将其转化至酿酒酵母INVSc1中表达,进行黄酮类底物粗酶检测。[结果]获得了大小约1 200 bp的EaPT1基因;经SDS-PAGE检测,重组质粒转化后的酵母菌有疑似EaPT1的表达产物;粗酶检测中未能检测到EaPT1酶活性。[结论]为粗毛淫羊藿植物次生代谢,特别是类黄酮生物合成途径的解析奠定基础。     

淫羊藿苷对鸡胚成骨细胞增殖和分化的影响

【研究目的】本文研究了淫羊藿苷对鸡胚成骨细胞增殖和分化的影响。【方法】将不同浓度的淫羊藿苷加入第二代成骨细胞中,分别于培养24、48、72h时,使用MTT法和PNPP法检测成骨细胞的增殖率和碱性磷酸酶(ALP)活性。【结果】淫羊藿苷促进鸡胚成骨细胞增殖和分化,但增殖作用不显著。其中当淫羊藿苷浓度为20ng/ml时,对成骨细胞增殖分化作用最强。【结论】淫羊藿苷能够显著促进鸡胚成骨细胞分化,对成骨细胞增殖有一定作用。     

Effects of icariin preconditioning on inflammatory responses in myocar-dial ischemia-reperfusion injury

AIM:To observe the effects of icariin on myocardial ischemia-reperfusion injury. METHODS:The left anterior descending coronary artery was ligated for 30 min and then loosened for 2 h to establish the rat model of myocardial ischemia-reperfusion injury. Forty-eight healthy adult male SD rats weighing 250~300 g were randomly divided into sham group, model group, low-, middle-and high-dose icariin groups, and aspirin group. The morphological changes of the myocardium were observed by HE staining. The protein expression of NF-κB p65 in the myocardial nucleus was determined by the method of immunohistochemistry. The content of tumor necrosis factor α (TNF-α) in the myocardial tissues was detected by Western blotting. The level of interleukin 1β (IL-1β) in the serum was measured by ELISA. The activity of myeloperoxidase (MPO) in the myocardial tissues was assayed by colorimetry. RESULTS:Compared with sham group, TNF-α content, IL-1β concentration, NF-κB expression and MPO activity in all other groups increased. Compared with model group, TNF-α content, IL-1β concentration, NF-κB expression and MPO activity in low-, middle- and high-dose icariin groups and aspirin group all decreased. No significant difference of the above parameters between high-dose icariin group and aspirin group was observed. CONCLUSION: Icariin preconditioning reduces inflammatory responses in the process of myocardial ischemia-reperfusion injury in a dose-dependent manner.     

HPLC法测定朝鲜淫羊藿中淫羊藿苷的质量分数

采用HPLC法对不同采收期、不同产地、不同海拔、不同药用部位的淫羊藿中淫羊藿苷的质量分数进行了测定．结果表明：淫羊藿苷在0．1～1．4g／L范围呈良好的线性关系;不同采收期的淫羊藿叶中淫羊藿苷的质量分数为0．31446％～3．68099％;临江产淫羊藿中淫羊藿苷质量分数最高;海拔629m的淫羊藿中淫羊藿苷质量分数最高;同一植株不同部位中淫羊藿苷的质量分数分布为叶〉地上部分〉茎．     

淫羊藿苷的超声波强化提取及其稳定性初探

以淫羊藿叶为原料,用正交试验法优化了淫羊藿苷的超声波提取工艺,所得最佳工艺条件为：提取温度50℃、提取溶剂60%甲醇溶液、液固比35∶1（mL/g）、超声波作用时间20 min。该法对淫羊藿苷的提取量与索氏提取法相当,但提取时间显著缩短,提取溶剂用量较少,不影响淫羊藿苷的稳定性,具有较高的可重复性。     

高效液相色谱法测定促孕灌注液中淫羊藿苷含量

以高效液相色谱法分离测定促孕灌注液中有效成分淫羊藿苷含量。采用Shim-PackVP-ODS(150 mm×4.6 mm,5μm)色谱柱,以甲醇-水(55∶45)为流动相;流速1 mL/m in,检测波长270 nm。淫羊藿苷分离良好,线性范围在0.1~4μg(r=1.000),平均回收率为99.9%,RSD为2.4%(n=6)。方法简便、准确,可以控制促孕灌注液的质量。     

鹿茸骨宝胶囊提取工艺的研究

[目的]筛选鹿茸骨宝胶囊的最佳提取工艺。[方法]采用正交试验设计,以淫羊藿苷作为考察指标,以加水量、提取次数、提取时间为因素优选鹿茸骨宝胶囊的提取工艺。[结果]鹿茸骨宝胶囊最佳提取工艺为取药材加10倍量水,煎煮3次,每次1.5 h。[结论]按照最佳提取工艺验证,淫羊藿苷提取率较高,该提取工艺可行。