Pharmacokinetics of 8-O-acetylharpagide and harpagide after oral administration of Ajuga decumbens Thunb extract in rats

Ethnopharmacological relevance

Ajuga decumbens Thunb is a medicinal plant native to China popularly used to treat chronic pelvic inflammation and hysteromyoma. Its main bioactive components are iridoid glycosides, such as 8-O-acetylharpagide and harpagide that had presented antibacterial, anti-inflammatory, and antiviral activities.

Aim of the study

To establish a sensitive LC–MS/MS method and compare the pharmacokinetics of 8-O-acetylharpagide and harpagide in rats after oral administration of their pure forms and from compounds obtained from Ajuga decumbens extract.

Materials and methods

Rats received orally 15 mg/kg (equivalent of 6 mg/kg 8-O-acetylharpagide and 1.5 mg/kg harpagide), 30 mg/kg and 60 mg/kg of Ajuga decumbens Thunb extract and were compared to animals that received 12 mg/kg of 8-O-acetylharpagide or 3 mg/kg of harpagide p.o. Concentrations of 8-O-acetylharpagide and harpagide in plasma were determined by LC–MS/MS method at different time points and all pharmacokinetic parameters were estimated by non-compartmental analysis.

Results

Results showed that the iridoid glycosides were quickly absorbed by oral route and showed a dose-dependence profile. Pharmacokinetic parameters of both glycosides were essentially the same except Tmax when dosed as the extract or pure forms.

Conclusion

8-O-acetylharpagide was metabolized to harpagide, which affected the pharmacokinetic profiles of harpagide when dosed as the extract. This pharmacokinetic study seems to be useful for a further clinical study of Ajuga decumbens Thunb extract.     

哈巴苷对病毒性心肌炎模型小鼠的影响

目的:本研究旨在探讨哈巴苷对病毒性心肌炎模型小鼠的作用。方法:将小鼠随机分为6组:对照组、柯萨奇病毒B3(coxsackievirus group B type3,CVB3)组、哈巴苷(低、中、高剂量)组和卡托普利组,采用CVB3诱导病毒性心肌炎小鼠模型。HE染色观察心肌病理损伤程度;紫外分光光度法检测肌酸激酶同工酶-MB(creatine kinase lsoenzyme-MB,CK-MB)、乳酸脱氢酶(lactatedehy drogenase,LDH)、心肌肌钙蛋白I(myocardialtroponin I,cTnI)和心肌钙蛋白T(cardiac troponin,cTnT)水平;超声心动图检测小鼠左室舒张期后壁厚度(LVPWd)、射血分数(EF%)、舒张早期二尖瓣血流速度(E)、舒张早期二尖瓣环运动速度(E′IVS);ELISA法检测血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β);蛋白免疫印迹法检测NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)、半胱天冬酶-1(cleaved caspase-1)的蛋白表达。结果:与CVB3组相比,哈巴苷低、中、高剂量组和卡托普利组心肌病理损伤程度明显改善;CK-MB、LDH、cTnI、cTnT水平明显降低(P<0.05);LVPWD、E/E′IVS明显降低(P<0.05),EF%明显升高(P<0.05);TNF-α、IL-6、IL-1β水平明显降低(P<0.05);NLRP3、cleaved caspase-1蛋白水平明显降低(P<0.05)。结论:哈巴苷对CVB3诱导的病毒性心肌炎小鼠具有治疗作用。     

15个产地玄参中哈巴苷与哈巴俄苷含量测定

目的 建立HPLC法同时测定玄参中哈巴苷、哈巴俄苷的含量。方法 采用HPLC-PDA法测定,固定相采用Eclipse Plus C₁₈色谱柱,以乙腈–0.03%磷酸水溶液为流动相,梯度洗脱;体积流量1.0 ml/min;检测波长210、280 nm。结果 哈巴苷、哈巴俄苷分别在0.1020～0.5100 mg/ml（r=0.9999）、0.0340～0.1700 mg/ml（r=0.9999）范围内线性关系良好,平均加样回收率分别为97.44%、97.08%,RSD分别为0.93%、1.24%。不同产地之间哈巴苷与哈巴俄苷含量差异大（P<0.01）,其中、四川陇东、浙江临安、磐安、河南南丰含量较高。结论 该方法稳定准确,重复性好,可用于玄参的质量控制。     

HPLC法测定养阴降糖胶囊中哈巴苷和哈巴俄苷的含量

目的：建立同时测定养阴降糖胶囊中哈巴苷和哈巴俄苷含量的方法。方法：色谱柱采用C18柱（4.6 mm×100 mm,5μm）;流动相为乙腈-0.03%磷酸,梯度洗脱;检测波长为210 nm;流速为1.0 ml/min。结果：哈巴苷、哈巴俄苷进样量分别在19.240~384.800 ng（r=1.000 0）、17.568~351.360 ng（r=1.000 0）范围内与峰面积线性关系良好,平均回收率分别为（98.33±1.85）%（RSD=1.88%）、（97.43±1.13）%（RSD=1.37%）。结论：该方法简单、准确、专属性强,可用于养阴降糖胶囊的质量控制。     

HPLC-ELSD法同时测定咳灵胶囊中的苦杏仁苷、桃叶珊瑚苷、哈帕苷、贝母辛、贝母素甲和贝母素乙

摘 要 目的：建立HPLC ELSD同时测定咳灵胶囊中的苦杏仁苷、桃叶珊瑚苷、哈帕苷、贝母辛、贝母素甲和贝母素乙含量的方法。 方法: 采用Ultimate XB C18色谱柱(250 mm×4.6 mm,5 μm),蒸发光散射检测器（漂移管温度105 ℃,氮气流速2.0 L·min-1）;以乙腈 甲醇（1〖KG*9〗∶〖KG-*2〗1）与0.4%醋酸溶液为流动相,进行梯度洗脱,流速：0.7 ml·min-1,柱温：35 ℃。 结果: 苦杏仁苷、桃叶珊瑚苷、哈帕苷、贝母辛、贝母素甲和贝母素乙的线性范围分别为13.56～271.20 μg·ml-1(r=0.999 2)、8.48～169.60 μg·ml-1(r=0.999 9)、4.89～97.80 μg·ml-1(r=0.999 7)、2.66～53.20 μg·ml-1(r=0.999 4)、1.82～36.40 μg·ml-1(r=0.999 8) 、2.04～40.80 μg·ml-1(r=0.999 6);平均加样回收率（RSD）分别为97.90%（1.20%）,99.21%（1.62%）,97.68%（0.75%）,98.36%（1.38%）,99.70%（0.79%）,97.95%（1.56%）（n=6）。结论: 本文建立的方法结果准确,重复性好,样品处理简便,可作为咳灵胶囊的质量控制方法。     

哈巴苷及哈巴俄苷对过氧化氢损伤血管内皮细胞的保护作用

目的探讨哈巴苷及哈巴俄苷对过氧化氢（H2O2）损伤人血管内皮细胞的保护作用及其机制。方法体外培养人脐静脉血管内皮细胞（HUVECS）,将生长良好的3代～5代细胞用于实验。实验分四组：对照组、H2O2组、哈巴苷干预组及哈巴俄苷干预组,光镜观察细胞形态,用四甲基偶氮唑蓝比色法（MTT法）检测细胞活力;取细胞上清液,比色法测定培养液中乳酸脱氢酶（LDH）和丙二醛（MDA）含量。结果哈巴苷及哈巴俄苷干预均使H2O2损伤的HUVECS形态趋于正常,活力增强,细胞膜损伤减轻,减少细胞LDH及MDA产生。结论哈巴苷及哈巴俄苷能对抗H2O2引起的损伤,保护血管内皮细胞。     

星点设计-效应面法优化玄参提取工艺

目的:优选玄参中哈巴苷和哈巴俄苷的提取工艺。方法:采用星点设计-效应面法,以哈巴苷与哈巴俄苷总提取率为指标,考察提取溶剂、液料比和提取时间对其的影响。结果:18%～32%乙醇,液料比20∶1～26∶1,提取30～48min时玄参总提取率最高。结论:星点设计-效应面法优化了玄参中哈巴苷与哈巴俄苷的提取工艺,预测性良好且简便可行。