Down-regulation of c-Src/EGFR-mediated signaling activation is involved in the honokiol-induced cell cycle arrest and apoptosis in MDA-MB-231 human breast cancer cells

Honokiol is a naturally occurring neolignan abundant in Magnoliae Cortex and has showed anti-proliferative and pro-apoptotic effects in a wide range of human cancer cells. However, the molecular mechanisms on the anti-proliferative activity in cancer cells have been poorly elucidated. In this study, we evaluated the growth inhibitory activity of honokiol in cultured estrogen receptor (ER)-negative MDA-MB-231 human breast cancer cells. Honokiol exerted anti-proliferative activity with the cell cycle arrest at the G0/G1 phase and sequential induction of apoptotic cell death in a concentration-dependent manner. The honokiol-induced cell cycle arrest was well correlated with the suppressive expression of CDK4, cyclin D1, CDK2, cyclin E, c-Myc, and phosphorylated retinoblastoma protein (pRb) at Ser780. Apoptosis caused by honokiol was also concomitant with the cleavage of caspases (caspase-3, -8, and -9) and Bid along with the suppressive expression of Bcl-2, but it was independent on the expression of Bax and p53. In addition, honokiol-treated cells exhibited the cleavage of poly (ADP-ribose) polymerase (PARP) and DNA fragmentation. In the analysis of signal transduction pathway, honokiol down-regulated the expression and phosphorylation of c-Src, epidermal growth factor receptor (EGFR), and Akt, and consequently led to the inactivation of mTOR and its downstream signal molecules including 4E-binding protein (4E-BP) and p70 S6 kinase. These findings suggest that honokiol-mediated inhibitory activity of cancer cell growth might be related with the cell cycle arrest and induction of apoptosis via modulating signal transduction pathways.     

HPLC测定不同厂家厚朴配方颗粒中厚朴酚与和厚朴酚的含量

摘 要 目的： 建立厚朴配方颗粒厚朴酚与和厚朴酚含量测定方法,比较不同厂家厚朴配方颗粒中厚朴酚及和厚朴酚的含量。方法: 应用HPLC法测定,色谱柱：HypersiL C₁₈(250 mm×4.6 mm,5 μm);流动相：A相为乙腈,B相为水,梯度洗脱;流速：1.0 ml·min^-1,柱温：25℃,检测波长：294 nm,进样量：20 μl。结果: 厚朴酚在0.873～26.190 μg·mL^-1范围内与峰面积呈良好的线性关系（r=0.999 5）,平均回收率为99.24%,RSD=2.00%（n=6）;和厚朴酚浓度在0.732～21.980 μg·mL^-1范围内与峰面积呈良好的线性关系（r=0.999 0）,平均回收率99.89%,RSD=1.33%（n=6）。不同厂家生产的厚朴配方颗粒中厚朴酚与和厚朴酚含量存在较大差异。结论:本方法简便、准确、重复性好,可用于厚朴配方颗粒中厚朴酚与和厚朴酚含量测定,不同厂家厚朴配方颗粒的质量差异提示规范药材种植、遴选、制定科学统一的配方颗粒生产工艺和质量控制标准的必要性。     

和厚朴酚抗小鼠急性哮喘气道炎症作用及可能机制探讨

目的探讨和厚朴酚对急性哮喘气道炎症的抑制作用,并分析其可能的机制。方法通过卵蛋白（VOA）刺激诱导小鼠急性哮喘模型,使用和厚朴酚对哮喘小鼠治疗后统计肺泡灌洗液中的中性粒细胞、淋巴细胞、巨噬细胞和嗜酸性细胞数及细胞总数,并通过 ELISA法检测肺泡灌洗液中的肿瘤坏死因子（TNF‐α）、白介素‐4（IL‐4）、IgE、Eotaxin的水平。同时取肺组织,制备组织切片,HE染色检测组织病理学的改变。此外,还通过Western blot法检测NF‐κBp65和I‐κb的表达变化。结果和厚朴酚有效的降低了肺泡灌洗液中的总细胞数（30976．00±3865．54 v s 64861．67±7613．98）,并同时降低了中性粒细胞、淋巴细胞、巨噬细胞和嗜酸性的细胞数,治疗前后相比较差异均具有统计学意义（ P ＜0．05）。ELISA检测结果也显示,和厚朴酚有效的抑制了TNF‐α、IL‐4、IgE、Eotaxin的表达,差异均具有统计学意义（ P＜0．01）。病理学实验则显示,与模型组大面积的组织损伤和炎性细胞富集细胞相比,和厚朴酚治疗后,组织损伤得到有效缓解,炎性细胞富集减少。NF‐κBp65和I‐κb的表达分析显示,与模型组NF‐κBp65大量表达,I‐κb表达减少相比,和厚朴酚治疗组N F‐κBp65表达被显著抑制,I‐κb表达显著增加,其差异均有统计学意义（ P ＜0．05）。结论和厚朴酚能够显著抑制急性哮喘诱导的炎症反应和肺组织损伤,其可能的作用机制为抑制N F‐κBp65的活性。     

厚术胶囊质量标准研究

目的:建立厚术胶囊的质量标准。方法:采用薄层色谱(TLC)法对方中苍术、三七、白芍和山楂药材进行定性鉴别;采用高效液相色谱法测定厚朴中厚朴酚、和厚朴酚的含量。结果:TLC斑点清晰,阴性对照无干扰。厚朴酚与和厚朴酚的进样量分别在0.080～0.400μg(r=0.9999)、0.048～0.240μg(r=0.9999)范围内与各自峰面积积分值呈良好的线性关系;平均回收率分别为98.42%、98.34%,RSD分别为0.63%、0.90%(n=6)。结论:所建标准可有效控制厚术胶囊的质量。     

和厚朴酚对自发性高血压大鼠血清TNF-α作用研究

目的观察和厚朴酚对自发性高血压大鼠血清TNF-α水平的影响。方法和厚朴酚低、高剂量组分别灌胃和厚朴酚200mg/(kg·d)、400mg/(kg·d);卡托普利组给予卡托普利30mg/(kg·d);对照组给予相同容积溶媒。给药7W后ELISA法测血清TNF-α水平。结果和厚朴酚低、高剂量两组大鼠血清TNF-α水平明显低于空白对照组(P<0.05),分别为(84.07±6.91)pg/mL、(81.69±7.15)pg/mL,与对照组(97.81±7.82)pg/mL比较差异具有显著性。结论和厚朴酚能明显降低自发性高血压大鼠血清TNF-α水平,抑制自发性高血压大鼠高系统炎症。     

厚朴药材中厚朴酚与和厚朴酚的含量测定及化学模式识别

厚朴是我国传统中药,为木兰科植物厚朴或凹叶厚朴的干燥干皮、根皮及枝皮,具有燥湿消痰、下气除满之功效。此前已有很多文献采用气相指纹图谱法研究厚朴中挥发性成分的含量。采用甲醇超声提取、高效液相色谱法(HPLC)对厚朴有效成分(厚朴酚与和厚朴酚)进行了含量测定,用系统聚类分析对不同产地的样品进行了化学模式识别研究。     

正交试验优选健脾化食口服液提取工艺

目的:优选健脾化食口服液的提取工艺。方法:以干浸膏收率及和厚朴酚、厚朴酚含量为定量指标,以加水量、提取时间、醇沉浓度和提取次数为考察因素,用正交试验优选最佳提取工艺。结果:最佳提取工艺为加入药材30倍量的水,提取2次,每次1h,醇沉浓度为70%。结论:优选的最佳提取工艺稳定、可行。     

Antifungal activity of magnolol and honokiol

Two neolignan compounds, magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl, 1) and honokiol (5,5'-diallyl-2,4'-dihydroxybiphenyl, 2), were isolated from the stem bark of Magnolia obovata and evaluated for antifungal activity against various human pathogenic fungi. Compound 1 and 2 showed significant inhibitory activities against Trichophyton mentagrophytes, Microsporium gypseum, Epidermophyton floccosum, Aspergillus niger, Cryptococcus neoformans, and Candida albicans with minimum inhibitory concentrations (MIC) in a range of 25-100 microg/ml. Therefore, compound 1 and 2 could be used as lead compounds for the development of novel antifungal agents.     

厚朴药材道地性的研究

采自浙、闽、川、杜、湘、赣、鄂等厚朴主产区７省１１县药材样品,以及在浙江景宁种源试验林中的样品,用高效液相色谱法（ＨＰＬＣ）测定厚朴酚类含量。结果表明,衡量厚朴药材质量的厚朴酚、和厚朴酚、厚朴酚类总含量、厚朴酚与和厚朴酚的比值等４个太,在产地与相应种源测定定值存在显著的相关性,而且厚朴的外观性状与内在质量也有显著的遗传性。传统观点认为质量上乘的川朴（川都产厚朴）实际上应归功于地方的品种,即品种类型     

高效液相色谱法测定藿香正气丸(大蜜丸)中厚朴酚、和厚朴酚含量

目的建立藿香正气丸(大蜜丸)中厚朴酚、和厚朴酚的含量测定方法。方法采用高效液相色谱法,以ZORBAXSBC18色谱柱(5μm,46mm×150mm,SNUSCL012593),乙腈水(68∶32)为流动相,流速为10mLmin,检测波长为294nm,室温25℃。结果厚朴酚在018μg～35μg范围内呈良好的线性关系,该方法平均回收率为985%,RSD为15%;和厚朴酚在011μg～22μg范围内呈良好的线性关系,该方法平均回收率为1018%,RSD为27%。结论本方法简单、灵敏度高,可用于藿香正气丸(大蜜丸)中厚朴酚、和厚朴酚的含量测定。