滨蒿内酯对大鼠哮喘模型气道平滑肌增殖及PKC-α蛋白表达的影响

目的观察滨蒿内酯对哮喘大鼠气道壁平滑肌增殖及PKC-α（protein kinase C-α,蛋白激酶C-α）表达的影响,探讨滨蒿内酯治疗哮喘的作用机制。方法40只雄性大鼠随机分成4组：正常对照组（N组）、哮喘组（A组）、地塞米松治疗组（D组）及滨蒿内酯治疗组（S组）,以卵清蛋白致敏和激发方法建立哮喘大鼠动物模型并给予相应治疗。测定各组肺功能,检测支气管肺泡灌洗液（BALF）中白细胞总数及嗜酸粒细胞（EOS）计数;用免疫组化法检测PKC-α蛋白表达水平、光镜下测定平滑肌层厚度及内外径。结果大鼠致敏2周及诱发哮喘4周后,肺功能测定表现为呼气时间延长,呼气峰压明显增大;A组BALF中的白细胞总数及Eos计数均较N组明显升高。哮喘模型组气道壁PKC-α蛋白含量和平滑肌层厚度较正常组均显著增加（均为P〈0.001）,气道内外径比值较正常组减小（P〈0.001）。气道壁平滑肌层厚度与PKC-α蛋白表达呈正相关（rs=0.771,P〈0.01）。结论滨蒿内酯能减轻哮喘大鼠气道壁平滑肌层的增厚并抑制PKC-α的表达。     

Effect of scoparone on neurite outgrowth in PC12 cells

The neurite outgrowth-promoting effects of scoparone isolated from the stem bark of Liriodendron tulipifera were investigated in PC12 cells. At a concentration of 200 microM, scoparone markedly induced neurite outgrowth from PC12 cells. Scoparone at 200 microM also enhanced the outgrowth of neurites from cells in the presence of nerve growth factor (NGF, 2 ng/ml). The levels of intracellular cyclic AMP and concentration of Ca2+ were also increased by 200 microM scoparone. In addition, scoparone at 200 microM increased the activities of extracellular signal-regulated protein kinase (ERK), cyclic AMP-dependent protein kinase (PKA), protein kinase C (PKC) and Ca2+/calmodulin kinase II (CaMK II). However, scoparone-induced neurite outgrowth was blocked by a mitogen-activated protein kinase inhibitor (U0126), a PKA inhibitor (H89), a PKC inhibitor (GF109203X) and a CaMK II inhibitor (KN62). These kinase inhibitors also reduced the scoparone-induced neurite outgrowth associated with NGF. These results suggest that scoparone can induce neurite outgrowth by stimulating the upstream steps of ERK, PKA, PKC and CaMK II in PC12 cells.     

滨蒿内酯鉴别迭鞘石斛可行性的探讨

目的：探讨迭鞘石斛薄层鉴别方法的可行性。方法：以HPLC法DAD扫描确认迭鞘石斛中是否含有滨蒿内酯。结果：在迭鞘石斛薄层鉴别试验中,供试品色谱在与对照品色谱相应的位置上有斑点,但斑点颜色与滨蒿内酯对照品的颜色有差异;在HPLC鉴别试验中,滨蒿内酯检测限为2.14 ng时,迭鞘石斛供试品中出现与滨蒿内酯相应的色谱峰,但它...     

滨蒿内酯与呼吸道变应性疾病

滨蒿内酯是从茵陈中提取的有效成分,有利胆保肝、消炎镇痛、降血脂、抗肿瘤、平喘、免疫调节等作用。近年来发现其在抗变态反应方面的作用可能会使其临床价值得到进一步提高。本文从提取、化学合成、免疫调节作用机制及对呼吸道变应性疾病的作用等方面对其进行综述。     

Differences in the effects of model inducers of cytochrome P450 on the biotransformation of scoparone in rat and hamster liver

The hamster is known to display very high rates of monooxygenase-mediated biotransformation. In comparison with other species little knowledge has been gathered with respect to the nature of its cytochrome P450 enzymes and their respective inducibility. We studied the consequences of induction of P450 enzymes in rats and Syrian golden hamsters using the regioselective oxidative O-demethylation of the coumarin derivative scoparone. This metabolic conversion indicates differential effects of P450 inducers in the rat, in which various types of inducers cause different shifts in the isoscopoletin/scopoletin metabolite ratio (I/S-ratio). Liver microsomes from hamster not treated with P450 inducers oxidized scoparone much more efficiently than liver microsomes of untreated rats. In rat liver microsomes total demethylation rates of scoparone increased upon in vivo treatment with phenobarbital or ß-naphthoflavone. Phenobarbital reduced the I/S-ratio whereas ß-naphthoflavone caused an increase in this ratio. In hamster liver microsomes both phenobarbital and -naphthoflavone treatments resulted in a decrease in the I/S ratio. In this species the total scoparone demethylation rate was not much affected by phenobarbital, but -naphthoflavone caused a huge increase in over-all scoparone biotransformation. In both species, dexamethasone, isoniazid and clofibrate were much less effective. In contrast to the rat, in the hamster the scoparone biotransformation profile cannot be used to differentiate between phenobarbital- or -naphthoflavone-treated animals.     

6,7-二甲氧基香豆素对K562细胞红系分化的影响

摘要：目的 探讨6,7-二甲氧基香豆素(6,7-dimethoxycoumarin,6,7-DOC)对K562细胞红系分化的影响及作用机制。方法：选择K562细胞作为研究对象,用联苯苄胺染色,考察1 μmol·L-1、5 μmol·L-1、10 μmol·L-1 6,7-DOC分别于0h、24h、48h、72h对K562细胞红系定向分化的联苯苄胺阳性率的影响;采用Western Blot检测不同浓度6,7-DOC诱导后K562细胞中红系分化标志蛋白α-globin、β-globin、γ-globin的表达变化;采用CCK-8和流式细胞术检测经6,7-DOC诱导后K562细胞的增殖和细胞周期分布情况。结果：5 μmol·L-1、10 μmol·L-1 6,7-DOC组联苯胺染色阳性率显著高于对照组,且呈时间与剂量依赖性;10μmol·L-1 6,7-DOC组α-globin、β-globin、γ-globin的蛋白表达量显著高于对照组;5 μmol·L-1和10μmol·L-1 6,7-DOC组两组细胞增殖率均显著低于对照组,且呈剂量、时间依赖性,6,7-DOC使K562细胞大量阻滞于G0/G1期,S期比例减少。结论：6,7-DOC能够抑制K562细胞增殖,使红系分化标志蛋白α-globin、β-globin、γ-globin表达增加,促进K562细胞向红系定向分化。     

蒿属香豆素对MCD饮食诱导的非酒精性脂肪性肝纤维化小鼠保护性作用的研究

目的:探讨中药茵陈蒿的主要成分蒿属香豆素对蛋氨酸胆碱缺乏(MCD)诱导的非酒精性脂肪性肝纤维化小鼠的作用及分子机制。方法:采用MCD饲料喂养小鼠6 周进行造模,将50只20 g左右的ICR雄性小鼠随机分为五组:空白对照组,模型组,丹酚酸B组(10 mg/kg),蒿属香豆素低剂量组(30 mg/kg),蒿属香豆素高剂量组(60 mg/kg)。造模后1周灌胃,5周后取小鼠肝脏组织和血清进行实验,对肝脏组织进行病理检查。通过试剂盒检测各组小鼠血清生化指标,比色法测肝脏组织丙二醛(MDA)、超氧化物歧化酶(SOD)含量。Western blot方法测小鼠肝脏组织液α-SMA,Ⅰ型胶原蛋白,E钙黏蛋白,波形蛋白、转化生长因子β₁(TGF-β₁)通路蛋白的表达。结果:不同剂量蒿属香豆素组和丹酚酸B组的NAS评分、MDA水平、α-SMA、Ⅰ型胶原、波形蛋白的表达明显降低,E钙黏蛋白、SOD水平显著增高(P<0.05),TGF-β₁及p-Smad2/3的表达降低,Smad7的表达明显增高(P<0.05)。结论:蒿属香豆素能抑制TGF-β/Smad通路,有效改善肝脏脂质沉积,降低小鼠肝脏组织内氧化应激水平,逆转上皮间质转化,减轻纤维化。     

HPLC法测定复方胆通胶囊中绿原酸和滨蒿内酯的含量

目的：建立测定复方胆通胶囊中绿原酸和滨蒿内酯的含量的高效液相色谱法。方法采用高效液相色谱法,Agilent Zorbax Extend-C18色谱柱（4.6mm ×250mm,5μm）44nm。结果绿原酸和滨蒿内酯分别在0.9168～9.1678μg· mL -1、0.4960～4.9600μg· mL -1范围内呈良好的线性关系（均为r＝0.9999）,平均回收率分别为99.12％、98.92％（n＝9）,RSD＝2.0％、RSD＝2.1％。结论本方法操作简便,结果准确、可靠。     

滨蒿内酯对大鼠哮喘模型蛋白激酶C-α基因表达影响的研究

目的观察滨蒿内酯对哮喘大鼠肺组织蛋白激酶C—α（proteinkinase Cα,PKC-α）mRNA及蛋白表达的影响,探讨滨蒿内酯治疗哮喘的作用机制。方法将40只雄性大鼠随机分成4组：正常对照组（N组）、哮喘组（A组）、地塞米松治疗组（D组）及滨蒿内酯治疗组（S组）。以卵清蛋白致敏和激发方法建立哮喘大鼠动物模型并给予相应治疗。测定N组及A组的肺功能,检测各组支气管肺泡灌洗液（BALF）中白细胞总数及嗜酸性粒细胞（Eos）计数;用逆转录一聚合酶链反应（RT—PCR）法检测肺组织中PKC—α mRNA,用免疫组化法检测其蛋白表达水平。结果大鼠致敏3周及诱发哮喘4周后,肺功能测定表现为呼气时间延长,呼气峰压明显增大;A组BALF中的白细胞总数及Eos计数、肺组织PKC—α mRNA及蛋白表达水平均较N组明显升高（P〈0．01）;滨蒿内酯治疗组哮喘大鼠BALF中白细胞总数和Eos计数较A组减少（P〈0．01）;PKC—dmRNA及蛋白表达水平较哮喘组下降,但仍较正常对照组升高（P均〈0．01）,D组与S组之间差异无统计意义（P〉0．05）。结论滨蒿内酯可显著抑制哮喘大鼠模型肺组织的PKC—α表达,可明显抑制哮喘大鼠气道炎症的发生发展,对哮喘有较好的治疗作用。     

Phytochemical constituents from the fruits of Acanthopanax sessiliflorus

Six compounds were isolated from the fruits of Acanthopanax sessiliflorus. Their structures were elucidated as (-)-sesamin, scoparone, protocatechuic acid, ursolic acid, hyperin and 5-hydroxymethylfurfural by physicochemical and spectroscopic analysis. Among them, scoparone, ursolic acid and 5-hydroxymethylfurfural were isolated for the first time from Acanthopanax species.