Glycation inhibitory activity and the identification of an active compound in Plantago asiatica extract

The glycation reaction involves a series of non-enzymatic reactions between the carbonyl group on reducing sugars and the amino group on proteins leading to the formation of advanced glycation end-products (AGEs), which are acknowledged to be involved in the pathogenesis of diabetic and aging-related complications. Consequently, the development of AGE inhibitors is considered to have therapeutic potential in patients with diabetes or age-related diseases. The preliminary results showed that a methanol extract (PAE) of Plantago asiatica, which is traditionally used as a folk medicine in Asian countries to treat fever, cough, wound etc., had strong glycation inhibitory activity. The effects of the extract on AGE fluorescence were dose-dependent, reaching 41% inhibition at 0.1 microg/mL of extract. The purified principle from PAE was identified as plantamajoside. As well as antioxidant activities, in vitro glycation inhibitory activities with 10 and 25 mm plantamajoside were higher than those with 10 and 25 mm aminoguanidine. The results demonstrate that PAE and plantamajoside had significant effects on in vitro AGE formation, and the glycation inhibitory activity and antioxidant activity of plantamajoside were comparable to those obtained using millimolar concentrations of the standard antiglycation agent aminoguanidine, and the antioxidant ascorbate, respectively.     

HPLC法测定复方前列舒丸中6个活性成分

目的建立HPLC法同时测定复方前列舒丸中大车前苷、柚皮苷、橙皮苷、新橙皮苷、朝藿定C、淫羊藿苷6个活性成分的方法。方法采用安捷伦Zorbax C18色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈–0.1%磷酸水溶液,梯度洗脱;检测波长为330 nm(0~20 min,测定大车前苷)、283 nm(20~40 min,测定柚皮苷、橙皮苷、新橙皮苷)、270 nm(40~50 min,测定朝藿定C、淫羊藿苷);柱温为35℃;体积流量为1.0 m L/min;进样量为5μL。结果大车前苷、柚皮苷、橙皮苷、新橙皮苷、朝藿定C、淫羊藿苷在10.11~202.20、49.62~992.40、15.46~309.20、44.62~892.40、18.39~367.80、52.18~1 043.60 ng与峰面积线性关系良好。大车前苷、柚皮苷、橙皮苷、新橙皮苷、朝藿定C、淫羊藿苷的平均回收率分别为97.4%、98.8%、99.5%、98.9%、97.2%、95.6%,RSD值分别为0.83%、1.2%、1.4%、1.5%、1.5%、1.0%。结论该方法稳定可靠、简便易行,同时测定复方前列舒丸中大车前苷、柚皮苷、橙皮苷、新橙皮苷、朝藿定C、淫羊藿苷6个特征性成分,为全面控制复方前列舒丸的质量提供了参考。     

Cytogenetic investigation of chromosomal aberrations in cells treated with plantamajoside from Plantago asiatica

Plantago asiatica is a member of the Plantaginaceae family, and is widely distributed in East Asia. In our previous work, a single active compound, plantamajoside was isolated and confirmed to have glycation inhibitory activity, and did not possess toxicity during a 90 day repeated oral toxicity test in rats. In the present study, a chromosomal aberration test was performed to investigate the genotoxicity of plantamajoside. From the results of the cytotoxicity test, plantamajoside proved to be less toxic when it was treated combined with S9 cell fractions. However, there was a significant increase in structural aberrations during the short‐term treatment of plantamajoside at its highest dose (5000 µg/mL) even when combined with S9. This seems to have been a natural phenomenon due to the very high dose of plantamajoside that was used. However, to confirm the safety of plantamajoside for its potential use as a phytochemical agent in health products, additional mutagenicity tests are necessary. Copyright © 2009 John Wiley & Sons, Ltd.     

HPLC同时测定虎贞痛风胶囊中大车前苷、虎杖苷、特女贞苷、白藜芦醇的含量

目的:建立同时测定虎贞痛风胶囊中大车前苷、虎杖苷、特女贞苷、白藜芦醇含量的HPLC方法。方法:采用高效液相法测定,色谱条件为Agilent SB-C18色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-0.1%甲酸溶液梯度洗脱,检测波长280 nm,流速1.0 m L·min-1,柱温30℃。结果:大车前苷、虎杖苷、特女贞苷、白藜芦醇分别在0.099 6~1.992(r=0.999 9),0.123~2.46(r=0.999 9),0.145~2.90(r=0.999 9),0.025~0.50μg(r=0.999 8)与峰面积呈良好的线性关系,平均回收率分别为99.27%,99.43%,98.83%,98.07%。结论:该方法快速简便,重复性好,专属性强,可作为虎贞痛风胶囊的质量控制方法。     

HPLC法测定前列康复胶囊中大车前苷、朝藿定C和淫羊藿苷

目的建立同时测定前列康复胶囊中大车前苷、朝藿定C、淫羊藿苷的HPLC法。方法安捷伦Zorbax C18色谱柱(250 mm×4.6 mm,5μm);以乙腈–0.1%磷酸水溶液为流动相,梯度洗脱;柱温35℃;检测波长为0~30 min,330 nm(测定大车前苷),30~50 min,270 nm(测定朝藿定C、淫羊藿苷);体积流量1.0 m L/min。结果大车前苷、朝藿定C、淫羊藿苷分别在10.08~201.60 ng、18.43~368.60 ng、52.25~1 045.00 ng线性关系良好;平均回收率分别为100.6%、97.2%、97.2%,RSD值分别为1.5%、1.3%、1.2%。结论该方法操作简单,重复性好,可有效的控制前列康复胶囊的质量。     

藏药大车前草中大车前苷的定性和定量分析

目的：建立藏药大车前草药材的定性定量方法。方法：以大车前苷为指标性成分,以硅胶G为吸附剂,乙酸乙酯-甲醇-甲酸-水（18：3：1．5：1）为展开剂,紫外光灯（365nm）下检测,建立大车前草药材的薄层色谱鉴别方法。用高效液相色谱法进行含量测定,DIKMADiamonsilC18 色谱柱（250x4．6mm5μm）;流动相为乙腈-0．1％磷酸（18：82）,流速：1mL／min;检测波长330nm;柱温：30℃。结果：大车前草药材薄层色谱在紫外光灯（365nm）下检识,大车前苷呈亮蓝色荧光斑点,斑点清晰,分离效果佳;高效液相色谱法含量测定大车前苷在0．09-11．22μg呈线性（r=0．999）;加样回收率为100．64％（RSD=0．95％）;测得4批大车前草中大车前苷的含量在0．094％-1．361％。结论：通过对4批大车前草中的大车前苷进行定性和定量分析,证明所建立的薄层色谱鉴别和高效液相色谱法含量测定方法简便可靠,准确度高,重复性好,为大车前草的质量标准建立提供了实验依据。     

热痹清颗粒的质量标准提升研究

目的：建立热痹清颗粒的质量标准。方法：采用薄层色谱法（TLC）对热痹清颗粒中知母、赤芍、忍冬藤、车前草进行定性鉴别；采用高效液相色谱法对热痹清颗粒中马钱苷、芍药苷进行含量测定，色谱柱为Wondasil C₁₈（4.6 mm×250 mm，5 μm），流动相为乙腈-0.2%磷酸水溶液梯度洗脱，流速为1.0 mL·min^-1，检测波长为236 nm（马钱苷）、230 nm（芍药苷），柱温为30℃。结果：TLC斑点清晰，分离度良好，阴性对照无干扰；马钱苷、芍药苷分别在各自进样量范围内与峰面积呈良好线性关系，相关系数均在0.999 9以上，平均加样回收率分别为100.04%和98.35%，RSD分别为1.46和1.64（n=6）。结论：该方法快速、准确、专属性强，可作为热痹清颗粒质量控制的方法。     

车前草中大车前苷的含量测定及提取工艺优选

目的 建立车前草中大车前苷的含量测定方法,并优选其提取工艺。方法 采用HPLC法测定大车前苷的含量,并通过正交设计考察乙醇浓度、用量及提取时间对车前草中大车前苷提取工艺的影响。结果 大车前苷在12.51~125.10 μg/ml浓度范围内,线性关系良好（r=0.999 6）,平均加样回收率为98.57%,RSD为1.45%;最佳提取工艺为10倍量60%乙醇提取2次,每次1 h。结论 本研究建立的含量测定方法简便、准确、重复性好;优选的提取工艺稳定可行。     

车前草中大车前苷的定性和定量分析

目的:研究、建立中药车前草药材的定性定量方法。方法:以大车前苷(plantamajoside)为专属性指标成分,以硅胶G为吸附剂,醋酸乙酯-甲醇-甲酸-水(18∶3∶1.5∶1)为展开剂,紫外光灯(365 nm)下检测,建立车前草药材的薄层色谱鉴别方法。用高效液相色谱法进行含量测定,艾杰尔Promocil C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%甲酸水(17∶83),流速1.0 mL.min-1,检测波长330 nm,柱温30℃。结果:车前草药材薄层色谱在紫外光灯(365 nm)下检识,大车前苷呈亮蓝色荧光斑点,斑点清晰,分离效果理想;高效液相色谱法含量测定大车前苷在0.049 9~11.966 4μg呈线性(r=0.999 9);加样回收率为100.6%,RSD 2.7%;测得11批车前草中大车前苷的含量在0.067%~1.80%。结论:通过对11批车前草中的大车前苷的定性和定量分析,证明所建立的薄层色谱鉴别和高效液相色谱法含量测定方法灵敏,准确可靠,重复性好,为车前草的品质评价和质量标准建立提供了科学依据。     

Healing effect of carboxymethyl chitosan-plantamajoside hydrogel on burn wound skin

BackgroundIt is known that hydrogels based on carboxymethyl chitosan (CMCS) have properties controling microbial growth, reducing inflammatory cell infiltration, and promoting collagen deposition. Plantamajoside (PMS), a natural Chinese herbal medicine with biological activity, has the properties of reducing inflammation, anti-oxidation, and promoting wound healing. However, the effects of carboxymethyl chitosan/plantamajoside hydrogel on partial thickness burn wounds remain unclear.MethodsThe healing effect of carboxymethyl chitosan/plantamajoside hydrogel was evaluated by in vitro cell viability assay, cell migration assay, and further evaluated in a rat model of partial-thickness burn wounds.ResultsThe hydrogels were highly porous with a pore size of about 250 μm, and these pores were interconnected. After adding plantamajoside, a dense microstructure was further formed. The hydrogels containing 0.25% plantamajoside significantly increased the viability and migration of L929 cells (P < 0.05). Carboxymethyl chitosan/plantamajoside hydrogel significantly improved wound healing, granulation tissue proliferation and re-epithelialization, and promoted collagen deposition (P < 0.05). Carboxymethyl chitosan/plantamajoside hydrogel also significantly decreased IL (interleukin)-1β, IL-6 and TNF-α expression, and increased IL-10 expression (P < 0.05). Furthermore, carboxymethyl chitosan/plantamajoside hydrogel significantly promoted the expression levels of VEGF, CD31, α-SMA (α-smooth muscle actin) and collagen III, and reduced the expression level of collagen Ⅰ (P < 0.05). Our data suggest that carboxymethyl chitosan/plantamajoside hydrogel promotes burn wound healing by accelerating angiogenesis and collagen deposition and reducing the inflammatory response.