响应面法优化连翘叶中连翘苷提取工艺研究

通过单因素试验研究乙醇浓度、料液比、提取温度、提取时间和提取次数5因素对连翘叶中连翘苷得率的影响.在单因素试验的基础上,采用四因素三水平的Box - Behnken Design (BBD)中心试验设计研究响应值以及最佳变量的组合,得出连翘叶中连翘苷提取的二次回归方程.通过响应面法综合评价提取连翘叶中连翘苷的最佳条件为80％乙醇、料液比1∶14、83℃、2h,在此条件下的连翘苷得率为0.3895％.     

鱼腥草芩蓝口服液指纹图谱研究

以鱼腥草芩蓝口服液为研究对象,摸索不同流动相、溶剂、柱温、流速条件下的色谱图,通过中药指纹图谱相似度软件评价系统,计算相似度等相关参数,最终确定一个HPLC指纹图谱条件：Thermo C18色谱柱（250 mm×4.6 mm,5 μm）,以甲醇（A）-1 %磷酸水溶液（B）为流动相梯度洗脱,柱温25 ℃,体积流量1.0 mL·min-1,检测波长为240 nm。本研究建立的方法可靠,简便易行,为全面控制鱼腥草芩蓝口服液的质量提供科学依据。     

Effects of phillyrin on VEGF and endostatin expression in Lewis lung carcinoma

AIM: To investigate the effects of phillyrin on vascular endothelial growth factor (VEGF) and endostatin expression in lung tumor tissues isolated from Lewis lung carcinoma. METHODS: The expression of VEGF and endostatin in control individuals and the patients with lung cancer was determined by immunohistochemistry. In the animal experiment, 5 groups of animals were examined: control, tumor model, and tumor model with 3 different concentrations of phillyrin treatments. For preparation of transplanted tumor model, Lewis cells were subcutaneously injected into the right limb armpit of the nude mice. After that, phillyrin was administered via oral gavage once daily for 20 d at dose of 5 or 10 g/kg, or twice daily at 10 g/kg. Lung tumor tissues isolated from each group were observed by hematoxylin-eosin staining. VEGF and endostatin expression were examined by immunohistochemistry. RESULTS: VEGF expression was increased in lung tumor tissues as compared with normal and pericarcinous tissues, while endostatin expression was decreased. Phillyrin significantly inhibited the tumor size and tumor tissue density dose-dependently, which was accompanied with a decrease in VEGF expression and an increase in endostatin expression. CONCLUSION: Phillyrin inhibits the development of lung tumor through reducing VEGF expression and increasing endostatin expression.     

Phillyrin produced by Colletotrichum gloeosporioides, an endophytic fungus isolated from Forsythia suspensa

To investigate the phillyrin-producing endophytic fungi from the medicinal plant Forsythia suspensa, a total of 24 strains of endophytic fungi were isolated from the healthy stems, leaves and fruits of the plant, 9 from stems, 5 from leaves, and 10 from fruits respectively. All fungal isolates were fermented in liquid PDA medium and their extracts were preliminary analyzed by TLC. One isolated strain G10, which was from the fruit of F. suspensa, had the same R_f value as authentic phillyrin. The potential phillyrin-producing fungus G10 was further analyzed by HPLC and HPLC-MS, and the results showed that the isolate G10 possessed of a retention time and ion peaks identical with the authentic compound phillyrin. The isolate G10 was identified as Colletotrichum gloeosporioides based on the morphological characteristics and ITS sequence analysis. The current research indicates that the numerous endophytic fungi inside the medicinal plants are precious resource for the pharmaceutical natural products that are originally from the plants.     

连翘有效成分对炎症模型的作用

目的:研究连翘提取物对细胞炎症模型的抗炎作用。方法:将连翘提取物按不同浓度作用于细胞炎症模型,观察细胞的形态变化,利用MTT法测定各孔的光吸收值。结果:连翘苷和连翘黄酮对照组细胞形态镜检观察未见明显变化,OD值略比空白对照组低;黄酮对照组细胞几乎全部破碎;不同浓度的大肠杆菌对照组细胞破裂、聚集成团,作用显著(P<0.05),且随着浓度的升高,对细胞形态的改变愈加显著。结论:连翘提取物对炎性细胞有不同程度保护,与同浓度菌液对照组相比,保护作用显著(P<0.05),在100μL浓度菌液中保护作用尤甚,具有一定的抗炎作用。     

膜分离技术在双黄连泡腾片除杂工艺中的应用

为了考察不同孔径陶瓷膜对中药双黄连水提液除杂效果的影响,以双黄连水提液为研究对象,比较5种不同孔径的陶瓷膜(50、100、200、450、800 nm)对滤液性状、膜通量衰减、有效成分保留率及固形物去除率的影响;同时以双黄连泡腾片中黄芩苷、绿原酸和连翘苷含量为考察指标,对膜过滤除杂与传统醇沉除杂工艺进行比较。结果表明,金银花+连翘水提液经滤过孔径为450 nm的陶瓷膜过滤后,滤液澄清、透明,绿原酸、连翘苷转移率分别为90.14%和90.28%,固形物去除率为29.75%;滤过孔径为200 nm的陶瓷膜对黄芩水提调酸调碱液除杂所制备黄芩提取物中黄芩苷含量为83.03%;醇沉除杂所制备双黄连泡腾片中黄芩苷、绿原酸和连翘苷平均含量分别为42.08、1.92和3.11 mg/g,膜过滤除杂所制备双黄连泡腾片中黄芩苷、绿原酸和连翘苷平均含量分别为42.55、2.07和3.23 mg/g,且不同批次含量测定的RSD值均3%(n=3)。综上所述,膜分离技术除杂所制备双黄连泡腾片工艺稳定可靠、简便易行,且绿色、安全,显著降低生产成本,对工业化生产和推广具有指导意义。     

不同产地连翘中连翘苷含量差异比较

[目的]对不同产地连翘中的主成分连翘苷的含量进行比较。[方法]用薄层色谱法对连翘中主成分进行定性分析并用高效液相色谱法对不同产地的连翘中连翘苷含量进行测定。[结果]从连翘中分离得到一种化合物,经鉴定为连翘苷,并测定到来自山西的连翘中连翘苷含量接近河南的2倍,来自云南的连翘中连翘苷含量略低于河南,说明不同产地的连翘中连翘苷含量有明显的差别。其主要原因是由于连翘生产地生态环境的影响以及栽培管理措施的不同。[结论]试验结果对扩大连翘产区,保证连翘的质量和提高连翘临床应用等有着重要的意义。     

连翘苷含药血清的制备及测定

本项目分别对连翘进行不同方法的提取,得到其产物,再进行实验动物给药并制备含药血清,最后利用HPLC方法测定其含量。色谱条件：色谱柱PT-230AColume heater Apollo C18柱(150mm×4.6mm,5μm),流动相为乙腈-水 25：75,流速0.8ml/min,检测波长为235nm,柱温25 ℃。结果显示：煎剂组(B组)和粗粉组(C组)在5个时间点都测不出连翘苷的含量,提取物组(A组)只有在1.5小时可以测到含量极低的连翘苷,连翘苷纯品组(D组)在0.5、1、1.5、2、4小时的连翘苷含量分别为：0.205mg/ml, 0.266mg/ml、0,099mg/ml、0.046mg/ml、0 mg/ml。