Structural Characterization of an Alkali-Soluble Polysaccharide from Angelica sinensis and Its Antitumor Activity in Vivo

A novel alkali-soluble polysaccharide (AASP) was isolated from Angelica sinensis (Oliv.) Diels under aqueous alkali treatment, and its structural characterization and antitumor activity in Vivo were evaluated in present study. Results of HPGPC and IC revealed that AASP was a neutral polysaccharide containing Ara, Gal and Glc in the mole ratio of 1.00 : 2.26 : 24.43, with the average molecular weight of 4.7 kDa. Periodate oxidation, Smith degradation, methylation, FT-IR, and NMR analyses further demonstrated that a preliminary structure of AASP was proposed as follows: (1→3)-linked arabinose, (1→6)-linked galactose, and (1→), (1→4), (1→6), (1→3,6)-linked glucose with α- and β-configuration. In Vivo antitumor assays, AASP exhibited prominent antitumor effects on H22 hepatoma cells with an inhibitory ratio of 48.57 % and effectively protected thymuses and spleens of tumor-bearing mice. Besides, AASP displayed a proliferation stimulating activity of immunocytes (splenocytes, peritoneal macrophages and natural killer cells), and an auxo-action for cytokines release (TNF-α, IL-2 and IFN-γ), leading to the apoptosis of H22 solid tumors cells via G0/G1 phase arrested. The above data demonstrated that AASP holds great application potential to be a safe and effective antitumor supplement in the future.     

四川当归属8种植物果实和叶柄解剖学研究

通过对四川当归属植物果实的解剖研究表明,果实均背腹压扁,背棱槽油管多为1,合生面油管多为2,在少数种间有一定差异,如茂汶当归和紫花前胡合生面多于2;侧棱翅状,较果体宽,或与果体近等宽,也有的窄于果体;胚乳腹面微凹或平直．偶有凸出。果实的油管特征、侧棱翅发达程度和胚乳的形状等特征可以为种间关系提供参考。叶柄类型多样,包括圆环型、“西”型、星散型等。叶柄横切面的形状、近轴面沟槽的有无、髓腔的有无以及叶柄横切面上维管束的数目和排列类型等特征比较稳定,并且具有明显的种间差异,可以作为本属分类的重要依据。     

四川当归属六种植物的核型

从居群水平对四川当归属(AngelicaL.)6种植物的核型进行了研究,其中管鞘当归核型为首次报道。6个种的染色体均为2n=2x=22,多为中部和近中部着丝粒染色体,核型多为2A型,仅疏叶当归石灰窑居群为1A型;峨眉当归核型为2n=22=10m 2sm 10st,茂汶当归核型为2n=22=16m 4sm 2sm(SAT),当归核型为2n=22=14m 8sm,阿坝当归核型为2n=22=14m 8sm,管鞘当归核型为2n=22=12m 8sm 2sm(SAT),而疏叶当归不同居群的核型或多或少发生了不同程度的变异。参照Stebbins的观点,峨眉当归以核型不对称系数最高而在6个种中显示一定的核型进化性,疏叶当归不对称系数较低,且出现1A型核型因而体现出一定的原始性。但是,综合形态解剖、花粉性状以及核型特征可以看出,四川当归属植物各性状间具有进化的不同步性,这也表明,作为当归属分布频度中心之一的四川地区,同时也是当归属的分化中心之一。     

Computer-assisted, high-performance liquid chromatography with mass spectrometric detection for the analysis of coumarins in Peucedanum palustre and Angelica archangelica

A reversed-phase HPLC method with atmospheric pressure chemical ionisation MS detection has been developed for the separation and identification of coumarins in plants of Peucedanum palustre L. (Moench) and Angelica archangelica (L.) var. archangelica. The Turbo Method Development program was utilised to optimise the mobile phase with two organic solvents (acetonitrile and methanol) and two aqueous solutions (1.0% formic acid and 10 mM ammonium acetate). Optimisation of the solvent gradients for the method was performed with the aid of the DryLab program. Analyses were carried out using a Phenomenex Prodigy RP C18 column. Fifty-two peaks (14 of which were associated with coumarins) were separated in 30 min from extracts of P. palustre, and 48 peaks (15 associated with coumarins) from extracts of A. archangelica. A total of 21 different coumarin-type compounds were identified in the aerial and the underground parts of the title plants. Isopimpinellin and pimpinellin were found for the first time in P. palustre and were identified by comparison of retention times and MS data obtained following the analysis of pure standards. This is the first report of the coumarin composition of the umbels of P. palustre.     

当归多糖通过AKT/FOXO3通路调节免疫性卵巢早衰小鼠内分泌功能

为探讨当归多糖(angelica polysaccharide,AP)对免疫性卵巢早衰小鼠内分泌功能的影响及分子机制,本研究将60只雌性BALB/c小鼠分为正常组、模型组、AP低剂量(100 mg/kg)、中剂量(200 mg/kg)、高剂量(400 mg/kg)组和补佳乐阳性组,以小鼠透明带多肽为抗原,皮下多点注射建立免疫性卵巢早衰(premature ovarian failure,POF)小鼠模型,灌胃相应药物,连续治疗4周后检测各组小鼠卵巢指数,检测各组小鼠血液SOD和MDA水平;通过ELISA检测各组小鼠卵巢组织IL-1β和IL-6水平,Western blotting检测各组小鼠p-AKT和FOXO3蛋白表达水平。与空白组相比,模型组小鼠卵巢指数显著降低,与模型组比较,AP低、中、高剂量组和阳性组小鼠卵巢指数显著升高,差异具有统计学意义(p<0.01)。与空白组比较,模型组小鼠血液SOD活性显著降低,MDA含量显著升高,与模型组比较,AP低、中、高剂量组和阳性组小鼠血液SOD活性显著升高,MDA含量显著降低。ELISA结果显示,与空白组比较,模型组小鼠卵巢组织IL-1β和IL-6含量显著升高,与模型组比较,AP低、中、高剂量组小鼠卵巢组织IL-1β和IL-6含量显著降低,差异具有统计学意义(p<0.01)。Western blotting结果显示,与空白组比较,模型组小鼠卵巢p-AKT和FOXO3蛋白表达显著降低,与模型组比较,AP低、中、高剂量组和阳性组小鼠卵巢p-AKT和FOXO3蛋白表达显著升高,差异具有统计学意义(p<0.01)。本研究结果说明,当归多糖AP激活AKT/FOXO3通路,抑制小鼠氧化应激,改善免疫性卵巢早衰小鼠内分泌功能,可在一定程度上抑制免疫卵巢早衰。     

超声波提取的当归多糖化学修饰及其抗氧化活性研究

采用超声波辅助法从当归中提取水溶性当归粗多糖(ASP),经过4种化学修饰分别得到硫酸化当归多糖(S-ASP)、磷酸化当归多糖(P-ASP)、乙酰化当归多糖(Ac-ASP)、羧甲基化当归多糖(C-ASP)。通过红外光谱对化学修饰前后ASP的结构进行表征,并进行抗氧化活性和清除自由基能力的测定,以获得一种抗氧化活性较强的当归多糖。结果显示:经化学修饰后的ASP分别具有相应的特征吸收峰,表明当归多糖的4种化学修饰均已成功;经化学修饰的4种当归多糖总还原能力均弱于未修饰多糖,且清除羟基自由基(·OH)的能力无明显变化,但清除1,1-二苯基-2-苦苯肼(DPPH·)自由基和抑制Fe2+诱发的脂质过氧化反应的能力有所增强,其中P-ASP清除超氧阴离子(O2·-)自由基的能力最强;Ac-ASP抑制Fe2+诱发的脂质过氧化反应的能力最强,且均呈现出一定的量效关系。本实验结果为当归多糖的进一步研究与开发利用提供了一定的科学依据。     

High-performance liquid chromatography with atmospheric pressure chemical ionization and electrospray ionization mass spectrometry for analysis of Angelica sinensis

An HPLC-PAD-API/MS method for analysing the chemical constituents of Angelica sinensis (A. sinensis) has been developed. ESI and APCI spectra, in both positive ion (PI) and negative ion (NI) modes, provided very useful information concerning the molecular weights of detected compounds. By comparing the retention times, UV spectra, mass spectra and molecular weights of detected compounds with those published in literature, 15 constituents of A. sinensis could be tentatively identified. This technique involving combined MS information may provide an objective, reliable and rapid analytical method for the quality control and database research of traditional Chinese medicines.     

Development of the CELLOP optimisation model for plant cell cultivation

A mathematical computer-aided model CELLOP was constructed in which the desirability functions in a three-dimensional experimental design can be used to find the optimal growing conditions for plant cells. CELLOP is intended for the optimisation of 3 to 6 physical, chemical, or biological variables in the cultivation conditions of plant cell cultures. The model was used to optimise the culturing conditions (calcium, inorganic nitrogen, and sucrose concentrations) for coumarin-producing, spontaneously embryogenic cell lines of angelica Angelica archangelica L. subsp. archangelica and hogs fennel Peucedanum palustre (L.) Moench. For A. archangelica the overall optimum concentrations were 0.47 mM Ca²⁺, 5.06 mM NO₃ ⁻, 0.40 mM NH₄ ⁺, and 96.25 mM sucrose. The dry mass was 24.7 % higher and the coumarin content 40.5 % higher than those achieved in the standard 75 % Gamborg B5 medium. For A. archangelica the highest embryogenic activity was reached in the media containing 1.25 mM Ca²⁺. In the case of P. palustre the overall optimum concentrations were 1.60 mM Ca²⁺, 2.84 mM NO³ ₋, 0.23 mM NH₄ ⁺, and 85.10 mM sucrose. For P. palustre the dry mass production increased by 61.8 % and the coumarin content by 58.1 % compared to the values achieved in the Gamborg B5 medium. For P. palustre the highest embryogenic activity was reached in the presence of 50.0 mM NO₃ ⁻ and 4.01 mM NH₄ ⁺.