HPLC法同时测定开胸顺气丸中橙皮苷、和厚朴酚与厚朴酚的含量

目的:建立同时测定开胸顺气丸中橙皮苷、和厚朴酚与厚朴酚含量的方法。方法:采用高效液相色谱法。色谱柱为Agi-lentZORBAXSB-C18柱,流动相为甲醇(A)-5%冰醋酸溶液(B)(梯度洗脱),流速为1.0mL·min-1,柱温为40℃,进样量为10μL,检测波长0～20min为284nm、20～65min为294nm。结果:橙皮苷进样量在0.1717～4.292μg范围内(r=0.9998)、和厚朴酚进样量在0.04004～1.001μg范围内(r=0.9994)、厚朴酚进样量在0.09280～4.640μg范围内(r=0.9998)与各自峰面积积分值均呈良好的线性关系;平均回收率分别为99.88%、99.34%、99.34%,RSD分别为0.67%、1.10%、0.86%(n=6)。结论:本法简便、准确、重复性好、精密度高,可更好地控制开胸顺气丸的质量。     

不同厂家藿香正气水质量比较研究及分析

目的:比较目前市场上流通的5个厂家藿香正气水的质量状况。方法:对5个厂家5批次药品进行装量差异和乙醇含量等常规检查,采用改进的高效液相色谱(HPLC)法测定药品中厚朴酚与和厚朴酚的含量,色谱柱为SHIMADZU HPLC-20AT ODSC18(150mm×4.6mm)柱,流动相为乙腈∶水=60∶40(2%冰乙酸),检测波长294nm。结果:改进HPLC方法测得和厚朴酚(r=0.9998)与厚朴酚(r=0.9999)在1～100mg·L-1范围内线性关系良好,厚朴酚的平均回收率为99.02%,RSD=1.78%;和厚朴酚的平均回收率为99.15%,RSD=0.97%。不同厂家生产的藿香正气水装量差异、乙醇含量、厚朴酚与和厚朴酚总量均符合2005年版《中国药典》中相关规定,但厚朴酚与和厚朴酚的含量差异较大。结论:建议进一步研究生产工艺,稳定原料药,并制定出科学、统一的质量标准。     

Honokiol induces paraptosis and apoptosis and exhibits schedule-dependent synergy in combination with imatinib in human leukemia cells

Honokiol, an active component isolated and purified from Chinese traditional herb magnolia, has been shown to inhibit growth and induce apoptosis in different cancer cell lines. This study shows that honokiol can induce a cell death distinct from apoptosis at lower concentrations. The death was characterized by cytoplasmic vacuolization with the endoplasmic reticulum swelling and accompanied by apoptosis at higher concentrations in NB4 and K562 cells. The two death processes may be in sequence at lower concentrations and in parallel with the increase of honokiol concentration. Membrane-associated cytotoxicity was involved in honokiol-induced paraptosis and apoptosis. Furthermore, honokiol inhibited concentration-dependent cell adhesion to extracellular matrix for NB4 cells. In addition, the cytotoxicity of honokiol combined treatment with imatinib was schedule- and concentration-dependent and the sequential administration of honokiol before imatinib appeared to be more beneficial in K562 cells. Taken together, the data suggest that honokiol induced a novel cell death pathway and there was cross-talk between apoptotic and non-apoptotic programmed cell death caused by honokiol in leukemia cells. Moreover, honikiol exhibited schedule-dependent synergy in combination with imatinib and sequential administration of imatinib followed by honokiol could be the optimal sequence to combine these two drugs in K562 cells.     

Preparation of honokiol-loaded chitosan microparticles via spray-drying method intended for pulmonary delivery

It has been demonstrated that spray-drying is a powerful method to prepare dry powders for pulmonary delivery. This paper prepared dispersible dry powders based on chitosan and mannitol containing honokiol nanoparticles as model drug. The results showed that the prepared microparticles are almost spherical and have appropriate aerodynamic properties for pulmonary delivery (aerodynamic diameters was between 2.8–3.3 μm and tapped density ranging from 0.14–0.?18?g/cm³). Moreover, surface morphology and aerodynamic properties of the powders were strongly affected by the content of mannitol. Fourier transform infra-red (FTIR) spectrum of powders indicated that the honokiol nanoparticles were successfully incorporated into microparticles. In vitro drug release profile was also observed. The content of mannitol in powders significantly influenced the release rate of honokiol from matrices.     

和厚朴酚在大鼠体内药动学及唾液分布研究

目的：探讨和厚朴酚静脉注射给药后在大鼠体内的动态变化规律及其在唾液中的分布情况。方法：大鼠尾静脉注射和厚朴酚,用反相高效液相色谱法测定和厚朴酚在血浆、唾液中的浓度,采用药动学软件3 P97分析数据,确定药动学参数。结果：和厚朴酚在大鼠体内符合二室模型分布,主要药动学参数：t1／2＝（102．980±10．600） min,AUC＝（1219．260±120．520）μg· min／mL,Vc＝（0．722±0．027） L／kg, CL＝（0．0246±0．002）L／min。结论：和厚朴酚可在唾液中检测到,给药后腮腺、颌下腺中的药物浓度与血浆中的药物浓度有一定的相关性。表明腮腺和颌下腺唾液样本将来可被用于和厚朴酚的治疗监测。     

和厚朴酚对人皮肤鳞癌细胞A431增殖凋亡作用及可能的机制分析

目的:研究和厚朴酚（Honokial,HNK）对人皮肤鳞癌细胞A431增殖凋亡作用及可能的机制分析。方法:采用CCK8实验方法检测和厚朴酚对A431细胞增殖的影响；Western blot定量分析和厚朴酚对Bcl-2和Bad蛋白表达的影响；Western blot定分析和厚朴酚对A431细胞中p53蛋白表达的影响；利用p53过表达腺病毒，并联合应用药物，通过CCK8和Western blot方法分析和厚朴酚对细胞增殖和凋亡的影响。结果:和厚朴酚以浓度和时间依赖性地抑制A431细胞增殖，并诱导其凋亡，上升Bad蛋白水平，下调Bcl-2蛋白水平；和厚朴酚可上调A431细胞中p53蛋白表达水平；加入p53过表达腺病毒可增强和厚朴酚的作用，抑制细胞增殖，促进细胞凋亡。结论:和厚朴酚能够抑制人皮肤鳞癌细胞A431的增殖并诱导凋亡，其机制可能与p53信号通路有关。     

和厚朴酚对组织因子途径抑制物 2诱导神经胶质瘤细胞凋亡的实验研究#br#

目的探讨和厚朴酚对组织因子途径抑制物 2（TFPI 2）表达的影响,以及和厚朴酚和TFPI 2过表达对人胶质瘤U251细胞凋亡的影响。 方法体外培养胶质瘤U87、U251、LN18、LN229、A172细胞系,荧光定量PCR（QPCR）检测不同细胞系中TFPI 2的表达。不同浓度（0、10、20、30、40和50 μmol／L）和厚朴酚干预U251细胞,QPCR和Western blotting检测TFPI 2 mRNA和蛋白的表达。采用腺病毒载体pGSadeno TFPI 2过表达U251细胞内TFPI 2的表达,流式细胞术及caspase 3试剂盒检测细胞凋亡率和caspase 3活性。 结果在选取的5种不同胶质瘤细胞株中,U251和A172细胞内TFPI 2 mRNA明显下降（P＜005）。和厚朴酚呈浓度依赖性增加U251细胞内TFPI 2 mRNA水平;其中50 μmol／L和厚朴酚干预24 h后, TFPI 2 mRNA表达水平升高最明显（P＜005）。腺病毒感染U251细胞后TFPI 2表达水平明显增加（P＜005）。过表达TFPI 2和50 μmol／L和厚朴酚干预均可增加U251细胞凋亡水平和caspase 3的活性（P＜005）。 结论和厚朴酚处理可增加U251细胞内TFPI 2的表达;和厚朴酚联合TFPI 2过表达显著诱导胶质瘤细胞凋亡。     

Anti-tumor effect of honokiol alone and in combination with other anti-cancer agents in breast cancer

Honokiol, an active component isolated and purified from Chinese traditional herb magnolia, was demonstrated to inhibit growth and induce apoptosis of different cancer cell lines such as human leukaemia, colon, and lung cancer cell lines; to attenuate the angiogenic activities of human endothelial cells in vitro; and to efficiently suppress the growth of angiosarcoma in nude mice. In this study, we have demonstrated that treatment of different human breast cancer cell lines with honokiol resulted in a time- and concentration-dependent growth inhibition in both estrogen receptor-positive and -negative breast cancer cell lines, as well as in drug-resistant breast cancer cell lines such as adriamycin-resistant and tamoxifen-resistant cell lines. The inhibition of growth was associated with a G1-phase cell cycle arrest and induction of caspase-dependent apoptosis. The effects of honokiol might be reversely related to the expression level of human epidermal growth receptor 2, (HER-2, also known as erbB2, c-erbB2) since knockdown of her-2 expression by siRNA significantly enhanced the sensitivity of the her-2 over-expressed BT-474 cells to the honokiol-induced apoptosis. Furthermore, inhibition of HER-2 signalling by specific human epidermal growth receptor 1/HER-2 (EGFR/HER-2) kinase inhibitor lapatinib synergistically enhanced the anti-cancer effects of honokiol in her-2 over-expressed breast cancer cells. Finally, we showed that honokiol was able to attenuate the PI3K/Akt/mTOR (Phosphoinositide 3-kinases/Akt/mammalian target of rapamycin) signalling by down-regulation of Akt phosphorylation and upregulation of PTEN (Phosphatase and Tensin homolog deleted on chromosome Ten) expression. Combination of honokiol with the mTOR inhibitor rapamycin presented synergistic effects on induction of apoptosis of breast cancer cells. In conclusion, honokiol, either alone or in combination with other therapeutics, could serve as a new, promising approach for breast cancer treatment.     

和厚朴酚神经保护作用的研究进展

和厚朴酚作为中药厚朴的主要活性成分之一,是一种多酚类化合物,具有抗炎、抗菌、抗肿瘤、抗氧化等多种作用.近年来发现,和厚朴酚对神经系统具有明显的保护作用.和厚朴酚易于通过血脑屏障,具有发展为广谱抗神经系统疾病药物的潜力.本文主要概述了目前和厚朴酚在神经保护方面的作用及机制,以期为后续的深入研究提供参考.     

Embryo-fetal development toxicity of honokiol microemulsion intravenously administered to pregnant rats

The aim of this study was to evaluate the embryo-fetal development toxicity of honokiol microemulsion. The drug was intravenously injected to pregnant SD rats at dose levels of 0, 200, 600 and 2000 μg/kg/day from day 6–15 of gestation. All the pregnant animals were observed for body weights and any abnormal changes and subjected to caesarean-section on gestation day (GD) 20; all fetuses obtained from caesarean-section were assessed by external inspection, visceral and skeletal examinations. No treatment-related external alterations as well as visceral and skeletal malformations were observed in honokiol microemulsion groups. There was no significant difference in the body weight gain of the pregnant rats, average number of corpora lutea, and the gravid uterus weight in the honokiol microemulsion groups compared with the vehicle control group. However, at a dose level of 2000 μg/kg/day, there was embryo-fetal developmental toxicity observed, including a decrease in the body length and tail length of fetuses. In conclusion, the no-observed–adverse-effect level (NOAEL) of honokiol microemulsion is 600 μg/kg/day, 75 times above the therapeutic dosage and it has embryo-fetal toxicity at a dose level of 2000 μg/kg/day, which is approximately 250 times above the therapeutic dosage.