Development of HPLC Method to Evaluate Drug-processing Technique of Eucommiae Cortex

Objective To establish a RP-HPLC method investigate the processing technique and mechanism of Eucommiae Cortex.Methods The RP-HPLC method was applied to simultaneously determining six ingredients,geniposidic acid,geniposide,genipin,chlorogenic acid,(+)-pinoresinol-di-β-D-glucopyranoside,and(+)-syringaresinol-di-β-D-glucopyranoside,in the different processed barks of Eucommia ulmoides.Results The valid method with good accuracy could be well used to study the processing technique of E.ulmoides;Besides,target ingredients in E.ulmoide were decreased within 6 h when they were processed.Conclusion Established RP-HPLC is a reliable method which could be used to research the processing technique of the barks of E.ulmoides.Moreover,the result of this study could be provided with significant evidence of processed barks of E.ulmoides.     

Effects of Eucommia ulmoides Extract on Longitudinal Bone Growth Rate in Adolescent Female Rats

Eucommia ulmoides is one of the popular tonic herbs for the treatment of low back pain and bone fracture and is used in Korean medicine to reinforce muscles and bones. This study was performed to investigate the effects of E. ulmoides extract on longitudinal bone growth rate, growth plate height, and the expressions of bone morphogenetic protein 2 (BMP‐2) and insulin‐like growth factor 1 (IGF‐1) in adolescent female rats. In two groups, we administered a twice‐daily dosage of E. ulmoides extract (at 30 and 100 mg/kg, respectively) per os over 4 days, and in a control group, we administered vehicle only under the same conditions. Longitudinal bone growth rate in newly synthesized bone was observed using tetracycline labeling. Chondrocyte proliferation in the growth plate was observed using cresyl violet dye. In addition, we analyzed the expressions of BMP‐2 and IGF‐1 using immunohistochemistry. Eucommia ulmoides extract significantly increased longitudinal bone growth rate and growth plate height in adolescent female rats. In the immunohistochemical study, E. ulmoides markedly increased BMP‐2 and IGF‐1 expressions in the proliferative and hypertrophic zones. In conclusion, E. ulmoides increased longitudinal bone growth rate by promoting chondrogenesis in the growth plate and the levels of BMP‐2 and IGF‐1. Eucommia ulmoides could be helpful for increasing bone growth in children who have growth retardation. Copyright © 2014 John Wiley & Sons, Ltd.     

杜仲水提液对幼鼠睾丸间质细胞睾酮合成和caspase9的影响

目的:探讨杜仲水提液对幼鼠睾丸间质(Leydig)细胞睾酮合成功能和caspase9的影响。方法:原代分离、鉴定幼鼠的Leydig细胞,进行细胞培养。在加入不同剂量杜仲水提液培养24 h后,用化学发光法测定睾酮的浓度;分别在加药24 h、36 h和48 h后,用MTT法检测各组的吸光度;在加药24 h后,用流式细胞仪分析细胞凋亡率、存活率和死亡率;用实时定量反转录聚合酶链反应测定caspase9 mRNA的表达。结果:在Leydig细胞中加入杜仲水提液24 h后,上清液的睾酮浓度明显增加,与空白组比较差异有统计学意义(P<0.01);药物刺激24 h、36 h和48 h后,其吸光度与空白组比较差异均有统计学意义(P<0.01);药物刺激24 h后,细胞凋亡率和死细胞率明显降低,活细胞率明显提高,与空白组比较差异有统计学意义(P<0.01);药物刺激24 h后,caspase9 mRNA的表达量明显降低,与空白组比较差异有统计学意义(P<0.05)。结论:杜仲水提液能通过下调caspase9的表达,降低Leydig细胞的凋亡,促进其增殖,而提高其合成睾酮的能力。     

正交试验优选杜仲多糖提取工艺

目的:优选杜仲多糖的提取工艺。方法:用普通水提醇沉法和微波提取法分别提取杜仲多糖,以提取温度、提取时间、料液比、提取次数为考察因素,以多糖提取率为评价指标,采用正交试验优选工艺。结果:普通水提醇沉法的最佳工艺为温度120℃、时间2 h、料液比1∶8、次数3次;微波提取法的最佳工艺为温度50℃、时间45 min、料液比1∶20、次数3次。结论:微波提取法对杜仲多糖的提取有辅助作用,较普通水提醇沉法更优。     

恩施产杜仲药材HPLC指纹图谱的研究

目的：建立恩施产杜仲药材水提液HPLC指纹图谱的分析方法。方法：10批恩施产杜仲药材水提,采用Wonda Sil C18（250 mm×4.6 mm,5μm）色谱柱,以乙腈-0.1%磷酸水溶液二元线性梯度洗脱,检测波长230 nm,流速1.0 ml·min^-1,柱温25℃,进样量10μl,进行HPLC指纹图谱分析。结果：确定了12个共有峰,采用《中药色谱指纹图谱相似度评价系统（2004A版）》计算10批杜仲药材水提液的指纹图谱整体相似度在0.596-0.997,并得出对照指纹图谱。结论：针对恩施产杜仲药材建立的HPLC指纹图谱分析方法,操作简便,稳定性高,重复性好,可有效地控制恩施产杜仲药材的内在质量。     

杜仲主要化学成分分类总结

本文综述了近年来国内外对中药杜仲主要化学成分的分类总结。为进一步研究杜仲药材的有效成分,控制其质量提供参考依据。     

杜仲酒抗疲劳作用的实验研究

〔目的〕 观察杜仲酒对小鼠抗疲劳能力的影响。〔方法〕 按卫生部《保健食品的功能学评价程序和检验方法》中抗疲劳检验方法进行测定。〔结果〕 高剂量组负重游泳时间明显高于对照组（Ｐ〈０．０１）;血清尿素氮中、高剂量明显低于对照组（Ｐ〈０．０５或Ｐ〈０．０１）;肝糖元高剂量组明显高于对照组（Ｐ〈０．０１）。〔结论〕 杜仲酒具有抗疲劳作用。     

杜仲叶提取物所含四种主要成分体外抗非酒精性脂肪肝活性研究＊

目的 研究杜仲叶所含4种主要成分体外抗非酒精性脂肪肝（Non-alcoholic fatty liver disease，NAFLD）活性。方法 首先用UPLC-MS检测杜仲叶提取物中绿原酸、杜仲苷、京尼平苷酸、车叶草苷含量；以HepG2细胞为载体，建立NAFLD细胞模型，设正常组、模型组、绿原酸组、杜仲苷组、京尼平苷酸组、车叶草苷组；油红O染色检测细胞内脂质；微量法检测细胞内甘油三酯（Triglyceride，TG）、胆固醇（Total cholesterol，TC）、高密度脂蛋白（High-density lipoprotein cholesterol，HDL-C）、低密度脂蛋白（Low-density lipoprotein cholesterol，LDL-C）含量；微量法检测细胞培养液中谷草转氨酶（Aspartate aminotransferase，AST）、谷丙转氨酶（Alanine aminotransferase，ALT）含量。结果 杜仲叶提取物主要含绿原酸、杜仲苷、京尼平苷酸和车叶草苷，其中京尼平苷酸的含量最高；杜仲叶提取物所含四种主要成分可明显降低细胞内TG、TC、LDL-C含量，其中京尼平苷酸效果最为显著；绿原酸、杜仲苷，京尼平苷酸可显著降低细胞培养液中AST、ALT含量，其中京尼平苷酸效果最好。结论 杜仲叶提取物所含四种主要成分可不同程度降低HepG2细胞内脂质含量，其中京尼平苷酸效果较好。     

中药杜仲炮制配伍的化学研究

目的：通过对中药杜仲炮制前后的化学成分研究得出其炮制的注意事项。方法：通过选择炮制方法,将三种方法炮制后的杜仲样品,分别采用UV光谱法、TLC法、HPLC法,分析杜仲生品和炮制品化学成分的差异。结果：通过炮制,杜仲的化学成分发生了改变。结论：在中医配伍的过程中必须考虑杜仲炮制中的化学成分变化。     

正交试验设计优化杜仲总黄酮提取工艺

目的:筛选出提取杜仲总黄酮的最佳工艺。方法:采用L9(34)正交试验设计,以提取时间、乙醇浓度、溶媒用量、提取次数为因素,采用紫外分光光度法对杜仲总黄酮含量进行检测,确定最佳提取条件。结果:最佳提取工艺为乙醇85%,溶媒用量16倍,提取90min,提取1次。结论:杜仲总黄酮提取工艺的优化,为其进一步研究提供参考。