不同海拔和光照对黄芩中7种黄酮类有效成分含量的影响

目的分析不同海拔、光照因素对黄芩中7种黄酮类有效成分量的影响,为黄芩种植最佳生长条件的选择提供理论依据。方法采用HPLC法同时测定人工种植的24批黄芩样品中野黄芩苷、黄芩苷、汉黄芩苷、芹菜素、黄芩素、汉黄芩素、千层纸素A7种成分的含量,并采用方差分析法进行不同海拔、光照环境(阴坡、阳坡)与各成分量的相关性分析。结果同一生长年限,在海拔(550±30)、(650±30)m 2个范围,对黄芩中7种成分的量进行比较,其中黄芩苷、汉黄芩苷有极显著性差异(P0.01),野黄芩苷、黄芩素、千层纸素A、汉黄芩素、芹菜素有显著性差异(P0.05);阴坡和阳坡2种不同的光照条件下,黄芩中7种成分的量相比,无显著性差异(P0.05)。结论在同一生长年限,随海拔的升高,黄芩中黄酮类成分的量都呈现明显的上升趋势,有显著性差异。而光照(阴坡、阳坡)不同时,黄芩中黄酮类成分的量无显著性差异,但除汉黄芩苷、黄芩素外,其他黄酮类成分含量的均数阳坡高于阴坡。黄芩人工种植宜选择阳光直接照射的平地或阳坡,同时及海拔偏高有利于提高黄芩中黄酮类化学成分总量的积累。     

止动颗粒UPLC指纹图谱建立及3种成分测定

目的建立止动颗粒UPLC指纹图谱,并测定其中3种成分的含量。方法以黄芩苷为参照峰,建立11批样品的UPLC指纹图谱,采用Acquity HSS T3色谱柱(150 mm×2.1 mm,1.8μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.3mL/min,柱温40℃,检测波长254nm,测定黄芩苷、栀子苷和芍药苷的含量。结果建立了UPLC指纹图谱,鉴定了没食子酸、车叶草苷、去乙酰车叶草苷酸甲酯、绿原酸、京尼平1-β-D-龙胆双糖苷、栀子苷、芍药苷、白杨素-6-C-β-D-葡萄糖-8-C-α-L-阿拉伯糖苷、白杨素-6-C-α-L-阿拉伯吡喃糖-8-C-β-D-葡萄糖苷、白杨素-6-C-α-L-阿拉伯吡喃糖-8-C-β-D-葡萄糖苷异构体、黄芩苷、黄芩苷异构体、千层纸素A-7-O-葡萄糖醛酸苷、汉黄芩苷、汉黄芩素、五味子醇甲16个成分,11批制剂相似度达到0.98以上。黄芩苷、栀子苷、芍药苷分别在12.15～388.80μg/mL、6.52～209.00μg/mL、8.38～268.00μg/mL线性关系良好,平均加样回收率(RSD)分别为100.98%(3.04%)、100.49%(0.60%)、100.55%(2.73%)。11批制剂中黄芩苷为7.46～12.40 mg/g,栀子苷为7.01～13.27 mg/g,芍药苷为7.68～12.76 mg/g。结论该方法准确简单,稳定可靠,可用于止动颗粒质量控制。     

UPLC-Q-TOF-MS快速鉴定止动颗粒血浆及尿液中移行成分

目的采用UPLC-Q-TOF-MS对止动颗粒血浆及尿液中移行成分进行快速定性,为体内药效物质基础研究提供参考。方法 Waters UPLC HSS T3(150 mm×2.1 mm,1.8μm)色谱柱,0.1%甲酸水(A)-乙腈(B)梯度洗脱,体积流量0.3 mL/min,柱温35℃,采用电喷雾电离化源(ESI),MSE扫描模式进行正、负离子检测。结果通过与空白生物样品和对照品的提取离子色谱图、质谱碎片裂解信息比对,初步鉴定出45个血浆及尿液中移行成分,包括28个原型成分和17个代谢产物。结论建立的UPLC-Q-TOF-MS分析方法可为止动颗粒在血浆及尿液中移行成分的定性分析提供一种快速、简便、可靠的分析手段,为该制剂的药效物质基础研究奠定基础。     

HPLC双波长法同时测定炎热清片中6种有效成分的含量

目的 建立HPLC双波长法同时测定炎热清片中龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素和汉黄芩素6种成分的含量。方法 采用Agilent TC-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为甲醇∶0.2%磷酸水溶液（梯度洗脱）,柱温为35℃,流速为1.0 mL·min^-1,检测波长0~20 min为254 nm,同时检测龙胆苦苷、栀子苷、芒果苷,20~50 min为280 nm,同时检测黄芩苷、黄芩素、汉黄芩素。结果 龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素、汉黄芩素分别在18.92~189.2 ng（r=0.999 9）,107.44~1 074.4 ng（r=0.999 9）,9.82~98.2 ng（r=0.999 9）,767.68~7 676.8 ng（r=1.000 0）,100.94~1 009.4 ng（r=0.999 9）,49.84~498.4 ng（r=0.999 9）线性良好,平均回收率（n=9）分别为98.31%,98.35%,98.40%,98.03%,98.46%,98.24%,RSD分别为0.4%,0.3%,0.3%,0.4%,0.4%,0.3%。结论 该方法灵敏、简便、准确,可用于炎热清片的质量控制。     

The effects of baicalein or baicalin on the colloidal stability of ZnO nanoparticles (NPs) and toxicity of NPs to Caco-2 cells

Recent study suggested that the presence of phytochemicals in food could interact with nanoparticles (NPs) and consequently reduce the toxicity of NPs, which has been attributed to the antioxidant properties of phytochemicals. In this study, we investigated the interactions between ZnO NPs and two flavonoids baicalein (Ba) or baicalin (Bn) as well as the influence of the interactions on the toxicity of ZnO NPs to Caco-2 cells. The antioxidant properties of Ba and Bn were confirmed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, with Ba being stronger. However, the presence of Ba or Bn did not significantly affect cytotoxicity, intracellular superoxide or release of inflammatory cytokines of Caco-2 cells after ZnO NP exposure. When Ba was present, the cellular viability of Caco-2 cells after exposure to ZnO NPs was slightly increased, associated with a modest decrease of intracellular Zn ions, but these effects were not statistically different. Ba was more effective than Bn at changing the hydrodynamic sizes, Zeta potential and UV–Vis spectra of ZnO NPs, which indicated that Ba might increase the colloidal stability of NPs. Taken together, the results of the present study indicated that the anti-oxidative phytochemical Ba might only modestly protected Caco-2 cells from the exposure to ZnO NPs associated with an insignificant reduction of the accumulation of intracellular Zn ions. These results also indicated that when assessing the combined effects of NPs and phytochemicals to cells lining gastrointestinal tract, it might be necessary to evaluate the changes of colloidal stability of NPs altered by phytochemicals.     

黄芩苷在呼吸道黏膜慢性炎症性疾病中的抗炎作用及机制研究进展

黄芩苷是源于中药黄芩根部的黄酮类化合物,是黄芩的主要活性成分,其药理作用广泛,具有良好的抗炎、抗癌、抗病毒和抗氧化等作用。以慢性鼻窦炎、过敏性鼻炎和哮喘为主的呼吸道黏膜慢性炎症性疾病是影响人类日常生活的严重慢性疾病之一,其发病机制复杂,目前对于此类疾病的治疗易产生不良反应且有一定的局限性。中药成分黄芩苷因其抗炎效果好,不良反应小而具有开发为治疗呼吸道黏膜慢性炎症性疾病的药物潜能。本文对黄芩苷在呼吸道黏膜慢性炎症性疾病中的抗炎作用及机制研究进展进行总结和归纳,为其在相关疾病的临床应用提供理论依据。     

RGD环肽修饰的姜黄素/黄芩苷靶向共递送纳米脂质体的制备工艺优化及表征

目的 优化RGD环肽修饰的姜黄素（curcumin,Cur）/黄芩苷（baicalin,Bai）共递送靶向纳米脂质体（RGD-Cur/Bai-Lip）的制备工艺并进行表征评价。方法 采用乙醇注入法制备RGD-Cur/Bai-Lip,首先通过Plackett-Burman筛选实验确定超声时间、胆脂比、水合温度作为制备RGD-Cur/Bai-Lip的关键因素,然后采用Box-Behnken响应面法进行工艺优化,以姜黄素包封率、黄芩苷包封率、总载药量为考察指标,采用AHP-复相关系数法-拉开档次法进行多指标组合赋权以确定各指标权重系数并计算综合评分,从而确定RGD-Cur/Bai-Lip的最优处方及关键工艺参数。最后对其形态、粒径电位、稳定性、体外释放度及溶血性等进行表征评价。结果 优选得到的最佳工艺为胆脂比1：12,水合温度60℃,探头超声时间为10 min。RGD-Cur/Bai-Lip外观澄清透明,呈亮黄色,对光可视淡蓝色乳光,透射电子显微镜下观察形态圆整、分散均匀。平均粒径为（101.10±0.62）nm,多分散系数（PDI）为0.191 0±0.014 3,Zeta电位为（1.59±0.07）mV,姜黄素包封率为（94.28±4.51）%,黄芩苷包封率为（76.93±1.35）%,总载药量为（2.27±0.09）%。7 d内稳定性良好,无溶血性,并具有一定的缓释作用。结论 优化处方后制备的RGD-Cur/Bai-Lip包封率高,粒径分布均匀,7 d内较稳定,无溶血性,并具有一定的缓释作用,为RGD-Cur/Bai-Lip的后续抗肝纤维化体内外研究提供了制剂基础。     

装载黄芩苷的溶菌酶-低相对分子质量壳聚糖纳米凝胶的制备及表征

目的基于蛋白-多糖纳米凝胶的制备技术,通过低相对分子质量壳聚糖对溶菌酶的修饰以及对环境因素的调节筛选,控制溶菌酶的自组装行为,制备一种绿色环保的具备核壳结构的溶菌酶-低相对分子质量壳聚糖纳米凝胶,从而达到减小粒径,提高包封率和载药量的目的,使其更有利于肾靶向。方法利用Maillard反应将溶菌酶与低相对分子质量壳聚糖按不同比例合成溶菌酶-低相对分子质量壳聚糖枝接物,通过SDS-PAGE蛋白电泳来优选出产量最高的合成比例;对合成的产物进行内源荧光、紫外测定等表征,并通过2,4,6-三硝基苯磺酸(2,4,6-trinitro-benzenesulfonic acid,TNBS)法测定接枝率;以粒径为评价指标,对溶菌酶浓度,反应体系pH值、加热温度等条件进行筛选优化,得到空白纳米凝胶最佳合成条件;选取适宜的载药方法,得到了载黄芩苷纳米凝胶。结果合成纳米凝胶的最佳工艺为溶菌酶与低相对分子质量壳聚糖比例1∶2、反应体系pH值11、溶菌酶-壳聚糖接枝物质量浓度为0.6 mg/mL、加热温度71℃、加热时间51 min。得到的溶菌酶-低相对分子质量壳聚糖枝接物接枝率为(24.7±2.9)%,空白纳米凝胶的粒径范围为16～120 nm、平均粒径为49.02 nm、PDI为0.132,载黄芩苷纳米凝胶的包封率为(95.00±2.54)%、载药量为(17.00±1.26)%。结论通过Maillard合成了溶菌酶-低相对分子质量壳聚糖纳米凝胶,并找到了最优的合成工艺。制备的纳米凝胶包封率高,粒径小,分布均匀,缓释效果明显。     

黄芩苷衍生物的合成及其生物活性研究

目的为改善黄芩苷的溶解性,设计合成系列黄芩苷衍生物,并评价其抗脂质过氧化活性和抗肿瘤活性。方法以黄芩苷和环氧乙(丙)烷为原料,通过羟乙基化、羟丙基化、酯化反应合成了一系列黄芩苷衍生物;分别采用生化法和台盼蓝染色法测试目标化合物对小鼠脑脂质过氧化产物丙二醛(MDA)的体外抑制作用及对白血病细胞HL-60的生长抑制活性。结果合成了13个黄芩苷衍生物,其结构经1H-NMR和MS确证。其中,化合物3⁶、116、11¹3为未见文献报道的新化合物。化合物1、6对MDA的抑制作用显著,化合物713为未见文献报道的新化合物。化合物1、6对MDA的抑制作用显著,化合物7¹0、12具有中等强度的抑制HL-60生长作用。结论黄芩苷结构中的酚羟基是其抑制脂质过氧化产物MDA的活性基团,葡萄糖醛酸的6位游离羧基是能够增强化合物抗肿瘤活性的有效基团。     

Baicalin suppresses iron accumulation after substantia nigra injury: relationship between iron concentration and transferrin expression

Previous studies have shown that baicalin prevented iron accumulation after substantia nigra injury, reduced divalent metal transporter 1 expression, and increased ferroportin 1 expression in the substantia nigra of rotenone-induced Parkinson's disease rats. In the current study, we investigated the relationship between iron accumulation and transferrin expression in C6 cells, to explore the mechanisms of the inhibitory effect of baicalin on iron accumulation observed in Parkinson's disease rats. Iron content was detected using inductively coupled plasma-atomic emission spectroscopy. Results showed that iron content decreased 41% after blocking divalent metal transporter 1 and ferroportin 1 proteins. After treatment with ferric ammonium citrate of differing concentrations(10, 50, 100, 400 μg/mL) in C6 glioma cells, cell survival rate and ferroportin 1 expression were negatively correlated with ferric ammonium citrate concentration, but divalent metal transporter 1 expression positively correlated with ferric ammonium citrate concentration. Baicalin or deferoxamine reduced divalent metal transporter 1 expression, but increased ferroportin 1 expression in the 100 μg/mL ferric ammonium citrate-loaded C6 cells. These results indicate that baicalin down-regulated iron concentration, which positively regulated divalent metal transporter 1 expression and negatively regulated ferroportin 1 expression, and decreased iron accumulation in the substantia nigra.