多药耐药相关蛋白2对汉黄芩素及其主要II相代谢产物药动学特征的影响

目的 研究多药耐药相关蛋白2（multidrug resistance protein 2,MRP2）对汉黄芩素及其主要II相代谢产物药动学特征的影响。方法 FVB野生型和MRP2^-/-小鼠ig汉黄芩素β-环糊精包合物后,于不同时间点眼眶取血。采用超高效液相色谱-质谱联用技术检测汉黄芩素、汉黄芩苷、汉黄芩素-7-O-硫酸酯的血药浓度,采用DAS 2.0软件以非房室模型计算药动学参数。结果 3个成分的线性关系良好,准确度、精密度、稳定性、回收率等符合要求。汉黄芩素主要以II相代谢产物汉黄芩苷和汉黄芩素-7-O-硫酸酯存在,均在30 min内达到最大血药浓度。汉黄芩苷的达峰浓度（C_max）和药时曲线下面积（AUC_0~t）最高,汉黄芩素-7-O-硫酸酯次之,汉黄芩素最低。与FVB野生型组比较,MRP2^-/-组汉黄芩苷和汉黄芩素-7-O-硫酸酯的C_max和AUC_0~t显著增加（P<0.05、0.01）。结论 MRP2介导了汉黄芩苷和汉黄芩素-7-O-硫酸酯的外排过程,显著影响汉黄芩素的体内处置过程。     

BisGMA‐induced cytotoxicity and genotoxicity in macrophages are attenuated by wogonin via reduction of intrinsic caspase pathway activation

Bisphenol‐A‐glycidyldimethacrylate (BisGMA) is a frequently used monomer in dental restorative resins. However, BisGMA could leach from dental restorative resins after polymerization leading to inflammation in the peripheral environment. Wogonin, a natural flavone derivative, has several benefits, such as antioxidative, anti‐inflammatory and neuroprotective properties. Pretreatment of macrophage RAW264.7 cells with wogonin inhibited cytotoxicity which is induced by BisGMA in a concentration‐dependent manner. BisGMA induced apoptotic responses, such as redistribution of phosphatidylserine from the internal to the external membrane and DNA fragmentation, were decreased by wogonin in a concentration‐dependent manner. In addition, BisGMA‐induced genotoxicity, which detected by cytokinesis‐blocked micronucleus and single‐cell gel electrophoresis assays, were inhibited by wogonin in a concentration‐dependent manner. Furthermore, wogonin suppressed BisGMA–induced activation of intrinsic caspase pathways, such as caspases‐3 and ?8. Parallel trends were observed in inhibition of caspase‐3 and ?8 activities, apoptosis, and genotoxicity. These results indicate wogonin suppressed the BisGMA‐induced apoptosis and genotoxicity mainly via intrinsic caspase pathway in macrophages. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 176–184, 2016.     

仙鹤草的化学成分研究

目的 研究仙鹤草Agrimoniae Herba的化学成分。方法 利用硅胶柱色谱、大孔树脂柱色谱、Sephadex LH-20凝胶柱色谱进行反复分离纯化,根据化合物的理化性质和波谱数据鉴定其结构。结果 从仙鹤草中分离得到19个化合物,分别鉴定为β-谷甾醇（1）、三十二烷醇（2）、三十一烷醇（3）、十九烷酸（4）、十六烷酸（5）、二十烷酸（6）、二十七烷酸（7）、胡萝卜苷（8）、委陵菜酸（9）、银锻苷（10）、芹菜素-7-O-β-D-吡喃葡萄糖醛酸甲酯（11）、芹菜素-7-O-β-D-吡喃葡萄糖醛酸丁酯（12）、仙鹤草酚B（13）、汉黄芩素（14）、芹菜素（15）、山柰酚（16）、乌苏酸（17）、槲皮素（18）、异槲皮苷（19）。结论 化合物2～4、6、7、14均为首次从龙芽草属植物中获得。     

RP-HPLC同时测定大卫颗粒中绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素的含量

目的:建立RP-HPLC法同时测定大卫颗粒中绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素的含量。方法:采用Diamon-sil C18(200 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.05%磷酸溶液(B)为流动相进行梯度洗脱(0～10 min,10%A→15%A;10～15 min,15%A→25%A;15～25 min,25%A→28%A;25～35 min,28%A→35%A),柱温35℃,流速1.0 mL.min-1,检测波长230 nm,进样量20μL。结果:绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素5个成分质量浓度分别在2.80～28.0μg.mL-1(r=0.9992)、25.71～257.1μg.mL-1(r=0.9994)、0.71～7.1μg.mL-1(r=0.9995)、0.81～8.1μg.mL-1(r=0.9990)、0.50～5.0μg.mL-1(r=0.9992)的范围内与峰面积呈良好的线性关系;方法的平均回收率(n=9)分别为100.0%,99.0%,99.0%,99.9%,99.4%。结论:所建立的高效液相色谱测定法简便、准确,重复性好,专属性强,可用于大卫颗粒的含量测定质量控制。     

A wogonin-loaded glycyrrhetinic acid-modified liposome for hepatic targeting with anti-tumor effects

Abstract

Liver cancer has become one of the most common fatal cancers worldwide, with morbidity rates increasing each year. Wogonin (WG) is an attractive candidate for the development of new anti-cancer drugs. In this study, a novel glycyrrhetinic acid (GA)-modified WG liposome was developed for use in targeted anti-cancer therapy. Three types of WG preparations were investigated: free wogonin in solution (WG), passively targeted wogonin liposomes (WG-Lip) and GA-modified wogonin liposomes (GA-WG-Lip). The entrapment efficiency, size and zeta potential were measured. Cellular uptake, cytotoxicity, in vivo bio-distribution and anti-tumor efficacy were also investigated. Addition of GA to the liposomes did not diminish the high entrapment efficiency observed in the liposomes without GA. GA-WG-Lip showed the greatest uptake and had an IC₅₀ value 1.46 times higher than that of WG-Lip. The GA-modified liposomes rapidly accumulated in the liver with a long retention time, and also displayed a better tumor inhibitory ratio than that of the unmodified liposomes. Overall, the data indicated that use of the GA-modified WG liposomes conferred improvements in bio-distribution, accumulation at the tumor and therapeutic efficacy, perhaps due to increased receptor-mediated uptake of liposomes by liver-targeted cells. Together, these data show that GA-WG-Lip is a promising means of targeted therapy for liver cancer.     

UPLC同时测定银黄颗粒中4种黄酮类成分含量

目的:建立银黄颗粒中黄芩苷、汉黄芩苷、黄芩素和汉黄芩素的UPLC含量测定方法.方法:采用ACQUITY UPLC H-Class超高效液相色谱仪,ACQUITY UPLC BEH C18色谱柱(2.1 mm× 50 mm,1.7 μm);流动相甲醇-0.2％磷酸水溶液,梯度洗脱,流速1 mL· min-1,检测波长274 nm,柱温30℃.结果:黄芩苷、汉黄芩苷、黄芩素和汉黄芩素的线性范围分别为1.09 ～21.8,0.340～6.80,0.116 ～2.32,0.102～2.04 μg,加样回收率分别为100.8％(RSD 0.32％),97.1％ (RSD 0.95％),101.5％ (RSD 1.6％),98.7％(RSD 0.059％).结论:该方法简便、快速,分离效果好,可为全面控制银黄颗粒的质量提供参考.     

RP-HPLC同时测定槐角丸中4种有效成分的含量

 目的 建立同时测定槐角丸中槐角苷、黄芩苷、染料木素和汉黄芩素4种有效成分含量的HPLC方法。方法 采用Diamonsil C₁₈柱(4.6 mm×200 mm, 5 μm进行分离;流动相为乙腈-0.05%磷酸溶液,梯度洗脱(0～10 min,25%～37%乙腈;10～25 min,37% ～60% 乙腈;检测波长260 nm;柱温30 ℃;流速1.0 mL·min-1;进样量10 μL。结果 槐角苷、黄芩苷、染料木素和汉黄芩素的质量浓度与峰面积分别在11.6~116 μg·mL-1(r=0.999 6, n=6、33.5~335 μg·mL-1 (r=0.999 8, n=6、0.46~4.63 μg·mL-1 (r=0.999 8, n=6和1.54~15.4 μg·mL-1(r=0.999 7, n=6内呈良好的线性关系;平均加样回收率依次为99.3%、99.6%、99.6%和99.0%。RSD为1.1%、1.1%、1.4%和0.8%。结论 该方法快速、简便、重复性好,适合于同时测定槐角丸中槐角苷、黄芩苷、染料木素和汉黄芩素的含量。     

HPLC法同时测定芩连片中黄芩苷﹑黄芩素和汉黄芩素的含量

目的建立同时测定芩连片中黄芩苷、黄芩素和汉黄芩素含量的HPLC方法。方法采用Dia-monsil C18柱(200 mm×4.6 mm,5μm)分离,以甲醇-体积分数为0.4%的磷酸溶液为流动相梯度洗脱,流速为1.0 mL.min-1,检测波长为277 nm,柱温为35℃。结果黄芩苷、黄芩素及汉黄芩素的线性范围分别为36.8～368.0 mg.L-1(r=0.999 7)、1.75～17.5 mg.L-1(r=0.999 8)、1.50～15.0mg.L-1(r=0.999 9),黄芩苷、黄芩素、汉黄芩素的平均回收率分别为97.3%(RSD=1.8%)、101.1%(RSD=1.8%)、99.0%(RSD=2.0%)。结论 HPLC法可用于芩连片的质量控制。     

黄芩药材的高效液相色谱指纹图谱及黄芩苷、黄芩素、汉黄芩素含量的同时测定

目的: 建立黄芩药材的HPLC指纹图谱,并同时测定不同产地商品中黄芩苷、黄芩素和汉黄芩素的含量,全面评价黄芩药材质量。方法: 采用Aglient TC-C₁₈(4.6 mm×250 mm,5 μm)色谱柱。采用梯度洗脱,流速为1.0 ml·min^-1;检测波长280 nm;柱温30℃。指纹图谱共有模式的建立采用国家药典委员会"中药色谱指纹图谱相似度评价系统2004A版"进行处理分析。结果: 在指纹图谱研究中,标定了9个共有峰,建立了对照指纹图谱,10批药材与共有模式之间相似性良好,相似度均在0.9以上;在所建立的分析条件下,黄芩苷、黄芩素、汉黄芩素均达到了基线分离,且在线性范围内线性关系良好,3种成分的加样回收率分别为99.44%,99.71%和99.83%。结论: 本法灵敏、准确、简便,可用于黄芩药材的综合质量评价。     

HPLC测定清肝活血方中4种黄酮的含量

 目的 建立HPLC同时测定清肝活血方中葛根素、黄芩苷、黄芩素和汉黄芩素含量的方法。方法 采用HPLC进行分析,色谱柱为Agilent Zorbax Eclipse XDB-C₁₈柱(4.6 mm×250 mm, 5 μm),流动相为甲醇-0.5%甲酸水梯度洗脱,流速为1.0 mL·min-1,检测波长为276 nm。结果 葛根素的线性范围为0.12~1.98 g·L-1(r=0.999 8),平均回收率为99.87%,RSD=2.04%;黄芩苷的线性范围为0.24~3.82 g·L-1(r=0.999 9),平均回收率为99.74%,RSD=1.84%;黄芩素的线性范围为0.01~0.14 g·L-1(r=0.999 6),平均回收率为99.64%,RSD=1.96%;汉黄芩素的线性范围为0.007~0.12 g·L-1(r=0.999 5),平均回收率为100.83%,RSD=2.37%。结论 本方法操作简便,测定结果准确可靠,可用于清肝活血方的质量控制。