薄层扫描法测定首清颗粒中黄芪甲苷的含量

目的建立首清颗粒中黄芪甲苷含量的测定方法。方法采用薄层扫描法，硅胶G薄层预制板，氯仿-甲醇-水（13：6：2）10℃以下放置过夜的下层溶液为展开剂，测定波长530nm，参比波长700nm。结果黄芪甲苷点样量在0．22～0．73μg之间与吸收度积分值呈良好的线性关系，平均回收率为97．72％，RSD=0．69％。结论该法便捷、灵敏、准确，可作为首清颗粒的质量控制方法。     

正交试验法优选芪术颗粒提取工艺

目的:优选芪术颗粒的提取工艺,为该制剂的工业化生产提供参考。方法:以黄芪甲苷、芍药苷提取量为综合评价指标,通过正交试验考察加水量、煎煮时间和煎煮次数对芪术颗粒水提工艺的影响。以特女贞苷和盐酸小檗碱提取量的综合评分为指标,通过正交试验考察溶剂用量、提取时间和提取次数对芪术颗粒醇提工艺的影响。采用HPLC-ELSD测定黄芪甲苷含量,流动相乙腈-水(34∶66)。采用HPLC-DAD测定芍药苷、特女贞苷和盐酸小檗碱的含量,流动相分别为甲醇-0.05 mol·L-1磷酸二氢钾溶液(30:70),甲醇-水(32∶68)和乙腈-0.05 mol·L-1磷酸二氢钾溶液(50∶50),检测波长依次为230,224,345nm。结果:最佳水提取工艺为加12,10倍量水提取2次,每次2 h。最佳醇提工艺为加6倍量60%乙醇提取3次,每次2 h。结论:优选的提取工艺稳定可行,适合推广于芪术颗粒的工业化生产。     

不同产地蒙古黄芪中黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ和黄芪皂苷Ⅲ含量的比较与分析

目的：采用HPLC-ELSD法测定了16批不同产地蒙古黄芪中黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ、黄芪皂苷Ⅲ的含量。方法：使用DIKMA Diamonsil C₁₈（150 mm×4.6 mm,5 μm）色谱柱,乙腈-水为流动相,梯度洗脱,流速：1 mL·min^-1,柱温：30℃。ELSD检测器参数：漂移管温度90℃,空气流量2.8 L·min^-1。采用SPSS 16.0进行聚类分析。结果：黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ和黄芪皂苷Ⅲ在进样量分别为0.093 2-1.02 μg（r = 0.999 5）, 0.789-8.78 μg（r = 0.999 7）, 0.506-3.13 μg（r = 0.999 6）,0.016 1-1.38 μg（r = 0.999 2）范围内,平均回收率分别为 97.55%、98.61%、99.68%、98.58%,RSD分别为1.2%、1.3%、1.3%、1.2%。聚类分析结果表明,单一采用黄芪甲苷为指标的聚类分析无法区分各省份蒙古黄芪,而同时采用4种皂苷含量为指标,可将不同省份蒙古黄芪明显区分开。结论：本方法简单、快捷、准确,能够更加直接地反映出不同产地蒙古黄芪原药材的质量状况,为蒙古黄芪药材的质量控制研究提供了新思路。     

不同产地蒙古黄芪中黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ和黄芪皂苷Ⅲ含量的比较与分析＊

目的：采用HPLC-ELSD法测定了16批不同产地蒙古黄芪中黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ、黄芪皂苷Ⅲ的含量。方法：使用DIKMA Diamonsil C18（150 mm×4.6 mm,5μm）色谱柱,乙腈-水为流动相,梯度洗脱,流速：1 mL·min-1,柱温：30℃。ELSD检测器参数：漂移管温度90℃,空气流量2.8 L·min-1。采用SPSS 16.0进行聚类分析。结果：黄芪甲苷、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ和黄芪皂苷Ⅲ在进样量分别为0.0932-1.02μg（r =0.9995）,0.789-8.78μg（r =0.9997）,0.506-3.13μg（r =0.9996）,0.0161-1.38μg （r =0.9992）范围内,平均回收率分别为97.55%、98.61%、99.68%、98.58%,RSD分别为1.2%、1.3%、1.3%、1.2%。聚类分析结果表明,单一采用黄芪甲苷为指标的聚类分析无法区分各省份蒙古黄芪,而同时采用4种皂苷含量为指标,可将不同省份蒙古黄芪明显区分开。结论：本方法简单、快捷、准确,能够更加直接地反映出不同产地蒙古黄芪原药材的质量状况,为蒙古黄芪药材的质量控制研究提供了新思路。     

一测多评法测定芪白平肺颗粒中8种皂苷类成分

目的建立一测多评法(QAMS)测定芪白平肺颗粒中8种皂苗成分的分析方法。方法以芪白平肺颗粒为研究对象,以人参皂苷Rb_1为内参物,计算人参皂苷Re、人参皂苷Rg_1、人参皂苷Rf、人参皂苷Rc、人参皂苷Rb2、黄芪甲苷、人参皂苷Rd与人参皂苷Rb_1的相对较正因子(f),通过f计算芪白平肺颗粒中7种成分,并在10批芪白平肺颗粒中进行验证,比较该方法与外标两点法测定结果的相似度。结果人参皂苷Rg_1、Re、Rf、Rb_1、Rc、Rb2、Rd、黄芪甲苷分别在进样量为0.416 0~4.992、0.315 4~3.785 2、0.259 6~3.115、0.385 2~4.622、0.222 8~2.674、0.193 2~2.318、0.183 5~2.202、0.574 6~6.895μg线性关系良好,f值分别为1.244、1.075、1.133、1.090、1.071、1.070、0.967,且在不同条件下重复性良好。QAMS法的计算结果与外标两点法的测定结果无显著性差异,再验证结果也无显著性差异,实验所得的f值可信。结论建立的QAMS可以准确、快速地实现芪白平肺颗粒中多种皂苷类成分的定量测定。     

Astragaloside IV Attenuates Injury Caused by Myocardial Ischemia/Reperfusion in Rats via Regulation of Toll‐Like Receptor 4/Nuclear Factor‐κB Signaling Pathway

Myocardial ischemia/reperfusion (MI/R) injury, in which inflammatory response and cell apoptosis play a vital role, is frequently encountered in clinical practice. Astragaloside IV (AsIV), a small molecular saponin of Astragalus membranaceus, has been shown to confer protective effects against many cardiovascular diseases. The present study was aimed to investigate the antiinflammatory and antiapoptotic effects and the possible mechanism of AsIV on MI/R injury in rats. Rats were randomly divided into sham operation group, MI/R group and groups with combinations of MI/R and different doses of AsIV. The results showed that the expressions of myocardial toll‐like receptor 4 (TLR4) and nuclear factor‐κB (NF‐κB) were significantly increased, and apoptosis of cardiomyocytes was induced in MI/R group compared with that in sham operation group. Administration of AsIV attenuated MI/R injury, downregulated the expressions of TLR4 and NF‐κB and inhibited cell apoptosis as evidenced by decreased terminal deoxynucleotidyl transferase dUTP nick end labeling positive cells, B‐cell lymphoma‐2 associated X protein and caspase‐3 expressions and increased B‐cell lymphoma‐2 expression compared with that in MI/R group. In addition, AsIV treatment reduced levels of inflammatory cytokines induced by MI/R injury. In conclusion, our results demonstrated that AsIV downregulates TLR4/NF‐κB signaling pathway and inhibits cell apoptosis, subsequently attenuating MI/R injury in rats. Copyright © 2015 John Wiley & Sons, Ltd.     

BCL-2、Caspase-3、NF-KB在黄芪总苷诱导人白血病NB4细胞凋亡过程中作用

本研究探讨黄芪总苷（astragaloside,AST）对急性早幼粒细胞白血病（APL）NB4细胞株的凋亡诱导作用及其分子机制。采用AST处理人APL细胞株NB4,CCK-8比色法检测细胞增殖抑制率,FITC—Annexinv／PI双染流式细胞术检测细胞凋亡率;不同浓度AST作用NB4细胞后RT-PCR检测BCL-2mRNA表达,Westernblot检测NF-KB、BCL-2、caspase-3蛋白表达水平。结果表明,AST作用NB4细胞48h后,浓度依赖性地抑制NB4细胞的增殖,且随着AST的浓度增加,细胞凋亡率也由4．69％上升到40．85％。RT—PCR结果显示,BCL-2mRNA表达减弱;Westernblot结果显示,NF—KB及BCL-2蛋白表达减弱,caspase-3表达增强。结论：降低NF-KB、BCL-2表达并最终激活caspase-3的表达可能是AsT诱导NB4细胞株凋亡的机制之一。     

黄芪甲苷对镉致大鼠睾丸支持细胞相关蛋白表达及p38MAPK磷酸化的拮抗作用

目的：探讨镉对培养大鼠睾丸支持细胞相关蛋白表达、p38M A PK磷酸化及超微结构的影响和黄芪甲苷的保护效应。方法对照组、镉（50 mol/L ）处理组、镉（50 mol/L ）加黄芪甲苷（10 mg/L ）组的培养支持细胞用于超微结构观察、波形蛋白、E-钙粘连蛋白/-环连蛋白免疫组织化学及磷酸化p38M A PK检测。结果镉处理组支持细胞线粒体内室肿胀,脂滴堆积,内质网扩张和（或）空泡化,髓样结构形成,少许支持细胞出现凋亡,镉加黄芪甲苷组支持细胞超微结构改变较镉处理组轻;免疫组织化学显示镉处理组波形蛋白、E-钙粘连蛋白及-环连蛋白阳性产物较对照组明显减弱（P＜0．05）,镉加黄芪甲苷组阳性产物虽较对照组减少但明显高于镉处理组（P＜0．05）;镉处理组支持细胞内磷酸化P38MAPK阳性产物表达量较对照组明显增强,且有向细胞核移位趋势,镉加黄芪甲苷组阳性产物表达量明显少于镉处理组（P＜0．05）。结论镉致大鼠睾丸支持细胞超微结构损伤、细胞骨架蛋白及粘连蛋白破坏并增强 P38MAPK磷酸化;黄芪甲苷可拮抗镉的毒性,其保护效应可能与减少 P38MAPK的磷酸化等有关。     

复方肾华片对急性脑缺血再灌注损伤大鼠肾小管Na+-K+-AT酶的调节作用

Objective:To observe the effect of Compound Shenhua Tablet(复方肾华片,SHT) on the sodiumpotassium-exchanging adenosinetriphosphatase(Na~+-K~+-ATPase) in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury(RIRI).Methods:Fifty male Wistar rats were randomly divided into the sham surgery group,model group,astragaloside group[150 mg/(kg-d)],SHT low-dose group[1.5 g/(kg·d)]and SHT high-dose group[3.0 g/(kg·d)],with 10 rats in each group.After 1 week of continuous intragastric drug administration,surgery was performed to establish the model.At either 24 or 72 h after the surgery,5 rats in each group were sacrificed,blood biochemistry,renal pathology,immunoblot and immunohistochemical examinations were performed,and double immunofluorescence staining was observed under a laser confocal microscope.Results:Compared with the sham surgery group,the serum creatinine(SCr) and blood urea nitrogen(BUN) levels were significantly increased,Na~+-K~+-ATPase protein level was decreased,and kidney injury molecule-1(KIM-1) protein level was increased in the model group after the surgery(P0.01 or P0.05).Compared with the model group,the SCr,BUN,pathological scores,Na~+-K~+-ATPase,and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery(P0.05 or P0.01).And the SCr,BUN of the SHT low- and high-dose groups,and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group(P0.05).The localizations of Na~+-K~+-ATPase and megalin of the model group were disrupted,with the distribution areas overlapping with each other and alternately arranged.The severity of the disruption was slightly milder in three treatment groups compared with that of the model group.The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group.Conclusions:The SHT effectively alleviated RIRI caused by ischemic reperfusion,promoted the recovery of the polarity of renal tubular epithelial cells,and protected the renal tubules.The therapeutic effects of SHT were superior to those of astragaloside as a single agent.     

正交试验优选糖肾康颗粒提取工艺

目的:优选糖肾康颗粒水提取工艺。方法:采用L9(34)正交试验法,以浸膏得率、黄芪甲苷和蜕皮甾酮提取量为指标,对加水量、煎煮时间、煎煮次数进行优选。结果:最佳提取工艺为:加水量10倍,煎煮3次,每次1.0h。结论:优选的工艺合理,重复性好,样品质量稳定。