HPLC法测定不同厂家产二十五味珊瑚丸中羟基红花黄花素A的含量

目的:运用HPLC法测定不同厂家产的二十五味珊瑚丸中羟基红花黄花素A的含量。方法:采用Agilent Polaris NH2柱(250mm×4.6mm,5μm);甲醇26%-乙腈2%-磷酸水72%(p H=2.5)作为流动相;流速1.0m L/min;柱温25℃进行检测。结果:厂家B产二十五味珊瑚丸中羟基红花黄色素A含量最多,厂家C产二十五味珊瑚丸其含量相对较少;测定出的二十五味珊瑚丸中羟基红花黄色素A含量范围在1051.19～464.62μg/g之间。结论:五个厂家生产的二十五味珊瑚丸中羟基红花黄色素A含量存在较大差异。

应用RAPD技术研究阿尔卑斯山黄花茅居群内的遗传分化(英文)

应用RAPD技术 ,沿一个海拔梯度研究了阿尔卑斯山黄花茅自然居群的遗传变异和分化。实验表明 ,虽然在亚居群间有很少的亚居群独有遗传标记的存在 ,但通过RAPD资料的聚类分析、主因子分析以及相关分析观察到遗传分化沿海拔梯度发生 ,而且亚居群间的遗传分化和它们的海拔高度 (地理距离 )呈有意义的正相关。研究结果暗示 ,在阿尔卑斯山的高山 亚高山过渡区 ,亚居群间的海拔高度差别可能导致黄花茅开花和生长物候期的变化 ,而后者限制了亚居群间的基因流 ,从而引起居群内的遗传分化。

红花总黄素抑制血小板激活因子所致毛细血管通透性增加的研究

本文研究了红花总黄素（ＳＹ） 拮抗血小板激活因子（ＰＡＦ） 诱导的小鼠毛细血管通透性增加的作用。小鼠随机分为正常动物空白对照组（Ａ），模型动物对照组（Ｂ） ，模型动物大剂量（Ｃ） ，中剂量（Ｄ） ，低剂量（Ｅ） 药物组共五组（ｎ ＝ １２） 。Ａ，Ｂ组腹注ＮＳ，Ｃ，Ｄ，Ｅ组腹腔注射ＳＹ；３０ｍｉｎ 后，各组尾静脉注射伊文斯蓝；尾注后３０ｍｉｎ，Ａ组腿部肌肉注射ＮＳ，Ｂ，Ｃ，Ｄ，Ｅ组肌注ＰＡＦ造模；肌注３０ｍｉｎ 后处死小鼠。提取腿部肌肉中的染料，测定提取液６２０ｎｍ 的吸收度反映毛细血管通透性。比色结果：Ａ 组０－０８０ ±０－０４５ ，Ｂ组０－４５６ ±０－１５６（ 与Ａ组比较，Ｐ＜ ０－０１） ，Ｃ 组（１００ｍｇ／ｋｇ ＳＹ）０－２２２ ±０－０７１ ，Ｄ 组（７０ｍｇ／ｋｇ ＳＹ）０－３２２ ±０－１３５ ，Ｅ 组（５０ｍｇ／ｋｇ ＳＹ）０－３４２ ±０－１２９ ，（Ｃ，Ｄ，Ｅ，组与Ｂ 组比较依次为，Ｐ＜ ０－０１ ，０－０５ ，０－０５） ，提示ＳＹ 具有明显抑制ＰＡＦ诱发的小鼠毛细血管通透性增加的作用，并且这种作用随着ＳＹ

Hydroxysafflor Yellow A Promotes Vascular Endothelial Cell Proliferation via VEGF/VEGF Receptor

Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.

红花中羟基红花黄色素A的纯化工艺研究

对羟基红花黄色素A纯化进行了工艺研究。采用恒温水提法对红花进行浸提,X-5型吸附树脂分离,采用Sephadex LH-20柱层析对纯化工艺中的条件进行筛选,运用紫外分光光度计测定羟基红花黄色素A的含量。结果显示:纯化的最佳工艺条件为上样量为2%的柱体积,流速为1.0 mL/min,洗脱液浓度为50%的乙醇,径高比为1∶8。采用紫外可见分光光度法测得羟基红花黄色素A的含量为86.4%。此方法具有操作方便,纯度较高的优点。

HPLC法测定红花地上部分羟基红花黄色素A和木犀草素的含量

目的:建立红花地上部分中羟基红花黄色素A和木犀草素的含量测定方法。方法:采用HPLC法,色谱柱为Eclipse plusC18柱(5μm,4.6mm×150mm),流动相为甲醇-0.7%磷酸梯度洗脱,流速:1.0 ml/min,检测波长390nm,柱温30℃。结果:羟基红花黄色素A在0.03μg/ml^0.19μg/ml,(r=0.999 9);木犀草素在0.13μg/ml^0.80μg/ml,(r=0.999 5)范围内有良好的线性关系,平均回收率为99.99%,98.95%(n=6)。结论:该方法简便、快捷、准确、重复性好,可用于羟基红花黄色素A和木犀草素的含量测定。

Elicitation on Artemisinin Biosynthesis in Artemisia annua Hairy Roots by the Oligosaccharide Extract from the Endophytic Colletotrichum sp. B501

The oligosaccharide elicitor from the mycelial wall of an endophytic Colletotrichum sp. B501 promoted the production of artemisinin in Artemisia annua L. hairy root culture. When hairy roots of 22-day-old cultures (later growth phase) were exposed to the elicitor (20 mg/L) for 4 d, the maximum content of artemisinin reached 1.15 mg/g, a 64.29% increment over the control. The electron X-ray microanalysis disclosed the rapid accumulation of Ca 2+ in the elicited cortical cells of hairy root. The electronic microscope observation revealed the high electron density area in vacuole of elicited cells. During the first day of elicitation the peroxidase activity of hairy roots was improved sharply. Some cellular morphological changes including cell shrinkage, condensation of cytoplasm and nuclear fragmentation, coincident with the appearance of DNA ladders, were observed after the third day of elicitation. It was suggested that the oligosaccharide elicitor triggered the programmed cell death, which may provide the substance or chemical signal for artemisinin biosynthesis.

PhenylpropanoidGlycosidesandFlavonoidGlycosidesIsolatedfromBudsofBuddlejaoficinalisMaxim

从醉鱼草属植物密蒙花的花蕾中分离得到６个苯丙素酚甙类成分，分别鉴定为ｖｅｒｂａｓ－ｃｏｓｉｄｅ（１），ｃｉｓｔａｎｏｓｉｄｅＦ（２），β－ｈｙｄｒｏｘｙａｃｔｅｏｓｉｄｅ（３），ｐｏｌｉｕｍｏｓｉｄｅ（４），ｅｃｈｉｎａｃｏｓｉｄｅ（５），ｍａｒ－ｔｙｎｏｓｉｄｅ（６）．它们在体外实验中显示了一定的抗肿瘤活性。同时还分离得到两个黄酮甙：蒙花甙（７）和芹菜素－７－芦丁糖甙（８）。其中化合物（２），（３），（４），（８）为首次从醉鱼草属中分离得到。

红花黄素预处理保存离体心脏的实验研究

目的观察红花黄素预处理对兔离体心脏保存的影响。方法家兔分别用Krebs-Henseleit液灌注20min、红花黄素灌注20min、缺血5min再灌10min,反复3次。各组心脏均经历停灌30min(保温、保湿)、缺血自动停跳及再灌注60min复制全心缺血-再灌注模型。应用离体心脏Langendorff灌注模型观察各组的血液动力学、冠脉流出液心肌酶及分子生物学改变。结果红花黄素预处理组左心功能、冠脉流量恢复明显优于对照组,心肌酶明显低于对照组,心肌诱导型一氧化氮合酶(iNOS)高于对照组。结论红花黄素预处理对兔心肌有保护作用。

Increase of β -1, 3-Glucanase and Chitinase Activities in Cotton Callus Cells Treated by Salicylic Acid and Toxin of Verticillium dahliae

The different resistance of cotton (Gossypium hirsutum L.) cultivars to crude toxin of Verticillium dah/iae(VD) was correlated with the activities of chitinase and β-1, 3-glucanase in callus cells. The activities of chitinase and β-1, 3-glucanase in the callus cells treated with the VD-toxin were increased to the higher level at earlier time point in resistant cultivars than these in the susceptible cultivars. Exogenous salicylic acid (SA) induced the accumulation of chitinase and β -1,3-glucanase, which resulted in the resistance of callus cells to the VD. toxin. Western blot using a polyclonal antibody against β -1,3-glucanase identified 28 kD protein that was induced by VD-toxin, SA, or VD-toxin plus SA.

Prediction Model of Secondary Substances in Anthocyanins Synthesis of Purple Corn

The aim of this study was to assay the polyphenols,flavonoid,polyphenol oxidase and phenylalnine ammonialyase which were relative to the anthocyanins synthesis of purple corn. The optimization of multiple linear regression model of anthocyanins synthesis was y=4.383 86-0.205 45x1＋5.479 638x2＋0.195 575x4. According to standard partial regression coefficient testing,the result indicated that polyphenols content was negatively correlated with anthocyanins and the relative influence to anthocyanins synthesis was-42.7%; flavonoid content and activity of polyphenol oxidase were positively correlated with anthocyanins of purple corn and the relative influence to anthocyanins synthesis were 71.45% and 73.32% respectively. There was no positive correlation between the activity of phenylalnine ammonialyase and anthocyanins of purple corn. The establishment of multiple linear regression model of anthocyanins synthesis was to provide theory foundation of producing anthocyanins in laboratory.

Lutein and lutein esters in marigold flowers by high performance chromatography

Lutein and lutein esters in marigold flowers was quantitatively determined by high performance chromatography （HPLC） with ODS-C 18 column.A mixture of CH3CNCD*2CH3OHCD*2CH3COOCH2CH3 with volume ratio of 55∶1∶44 was used as mobile phase at a flow rate of 1.0 mL/min and detection was carried out at 460nm. The column temperature was about 20℃. The contents of lutein and lutein esters were determined by analytical curve of lutein since lutein and lutein esters have the same spectral characteristics. Determination results of hexane extracts and saponified samples of lutein show that the saponification transforms the esters into free lutein. The increase of the content of dipalmitate and palmitate stearate reveals that the reaction includes transesterifications.

Stability and degradation of hydroxysafflor yellow A and anhydrosafflor yellow B in the Safflower injection studied by HPLC-DAD-ESI-MS^n

Safflower is a popular Chinese medicinal plant and Safflower injection is extensively used for the clinical treatment of cerebrovascular and cardiovascular diseases. In this study, HPLC-DAD-ESI-MSn was utilized to study the stability and degradation of the two major but chemically unstable bioactive compounds hydroxysaffior yellow A and anhydrosaffior yellow B, in Safflower injection. The impact of light irradiation, temperature, and pH on the stability of these two compounds were studied. The results showed that hydroxysafflor yellow A and anhydrosafflor yellow B could degrade at high temperature （〉60 ℃） or extreme pHs （pH ≤ 3.0 or 〉7.0）, but not under light irradiation. The common degradation product was p-coumaric acid. Chemical structures of the other degradation products were characterized by LC-MS. Hypothetical degradation pathways were proposed. In addition, ADP-induced platelet aggregation tests showed that the degradation of anhydrosaffior yellow B could reduce the anticoagulation activities of Safflower injection. Our results suggest that temperature and pH are critically important for the preparation and storage of Safflower injection.

红花黄色素抗大鼠急性心肌缺血研究

目的:观察红花黄色素对大鼠实验性心肌缺血的影响。方法:大鼠舌下静脉注射垂体后叶素(Pit)1u/kg制备急性心肌缺血模型,观察红花黄色素对大鼠心肌缺血百分率的影响;大鼠皮下注射盐酸肾上腺素0.8mg/kg加冰水刺激复制大鼠急性血瘀证模型,观察红花黄色素对急性血瘀证大鼠血液流变性的影响;小鼠常压耐缺氧实验,观察红花黄色素对小鼠耐缺氧能力的影响。结果:红花黄色素18、9、4.5 mg/kg舌下静脉给药能显著对抗Pit致大鼠心肌缺血百分率;降低"血瘀症"模型大鼠的全血比黏度、血浆比黏度、红细胞压积及纤维蛋白原含量;明显延长小鼠常压耐缺氧时间。结论:红花黄色素对大鼠急性心肌缺血具有保护作用,其作用与降低缺血百分率,改善血液流变学和减少心肌耗氧等有关。

羟基红花黄色素A对猪冠状动脉环的舒张作用及机制

目的：研究羟基红花黄色素A（HSYA）对离体猪冠状动脉血管的舒张作用及其可能作用机制，为红花的生物学活性研究和应用提供参考。方法：以猪冠状动脉血管环为材料，以离体血管功能实验方法检测羟基红花黄色素A对猪冠状动脉血管活性作用。结果：羟基红花黄色素A对静息状态猪冠状动脉血管环无明显舒张作用；HSYA在10^-4.5～10^-2mol／L的浓度范围内对PGFh（10^-6mol／L）预收缩的冠状动脉血管环具有浓度依赖性舒张作用，最大舒张效应为124．22％±6．25％，相应的pD2值为2．91±0．23，HSYA的有效剂量为10^-3mol／L，该剂量能使PGF2α（10^-6mol／L）预收缩的冠状动脉血管环达到65．55％±4．15％的舒张效应；HSYA可使KCl的量效曲线下移，呈浓度依赖性。用一氧化氮合成酶抑制剂L—NNA、鸟苷酸环化酶抑制剂亚甲蓝、β受体阻断剂普萘洛尔、β1受体抑制剂atenolol和B2受体抑制剂ICI 118551预处理后，均可明显减弱HSYA诱导的舒张血管作用；前列腺素合成酶抑制剂吲哚美辛，K＋通道抑制剂TEA预处理后，血管舒张作用不能被阻断。结论：羟基红花黄色素A的舒血管活性作用通过内皮．NO．cGMP途径和β-肾上腺素受体途径，并能够阻断血管平滑肌上的电压依赖性钙通道，但与血管平滑肌舒张因子前列腺素的释放及钾离子（KCa）通道无关。

红花黄色素对Aβ_(1-42)诱导的痴呆大鼠脑组织炎症因子释放的影响

目的:观察红花黄色素(SY)对Aβ_(1-42)海马注射致痴呆大鼠学习及记忆的影响,探讨红花黄色素对痴呆大鼠脑组织中炎症因子释放的调节作用。方法:Wistar大鼠60只随机分为6组,假手术组、模型组、红花黄色素10mg/kg、30mg/kg、100mg/kg组、多奈哌齐0.5mg/kg组。建立Aβ_(1-42)双侧海马注射致痴呆大鼠模型,造模后连续给药4周,至行为学测试结束。Morris水迷宫实验和跳台实验检测痴呆大鼠的空间学习记忆能力的变化;HE染色观察大脑皮层形态学改变;试剂盒法测定大鼠海马、皮层组织中i NOS、IL-1β、IL-6和TNF-α的含量。结果:与假手术组比较,模型组大鼠学习记忆能力明显下降;白介素-1β、白介素-6、肿瘤坏死因子α和诱导型一氧化氮合酶水平明显增加。经红花黄色素治疗后,红花黄色素10、30、100mg/kg学习记忆能力显著改善;HE染色结果显示,红花黄色素10、30、100mg/kg可减少神经元丢失数量,改善神经元形态异常;其对皮层IL-1β、IL-6、TNF-α和海马IL-6、i NOS的表达下调有显著性差异,同时其对海马TNF-α的表达下调也有明显的差异。红花黄色素10、30mg/kg对皮层中i NOS的表达下调有显著差异,红花黄色素30,100mg/kg对海马中IL-1β的表达下调有显著差异。结论:红花黄色素对Aβ_(1-42)海马注射致痴呆大鼠学习记忆障碍有显著改善作用,可能与其减少炎症因子释放有关。

红花黄色素对Aβ1-42诱导的痴呆大鼠学习记忆能力的影响

目的:大鼠海马区注射寡聚态的Aβ1-42,观察红花黄色素对大鼠空间学习、记忆能力的影响。方法:采用Wistar大鼠双侧海马注射Aβ1-42各2.5μl制备痴呆大鼠模型,实验设置假手术组、模型组、加兰他敏3 mg/kg组、红花黄色素10 mg/kg、30 mg/kg、100 mg/kg组,造模5天后灌胃至行为学结束。水迷宫实验,检测大鼠空间学习、记忆能力的改变状况。酶标仪测定大鼠血清TCHE和皮层、海马中SOD、MDA、GSH-Px和T-CHE含量。结果:模型组逃避潜伏期较假手术组明显延长,穿越平台次数显著降低,出现明显的空间学习障碍;与模型组比较,红花黄色素10 mg/kg、30 mg/kg、100 mg/kg均可使大鼠皮层组织中SOD,GSH-Px含量增加,MDA含量显著降低。红花黄色素10 mg/kg使大鼠海马组织中SOD含量明显增加,红花黄色素10 mg/kg、30 mg/kg、100 mg/kg使MDA含量显著降低;红花黄色素10 mg/kg、30 mg/kg、100 mg/kg均可显著增加大鼠海马组织中T-CHE含量,血清中T-CHE含量变化不明显。结论:红花黄色素可能通过提高脑组织抗氧化能力、改善胆碱能神经系统损伤以缓解Aβ1-42致大鼠痴呆症状。

羟基红花黄色素A对大鼠心肌细胞缺血再灌注损伤的保护作用

目的:探讨羟基红花黄色素A对心肌缺血再灌注损伤(I/R)的影响。方法:采用大鼠冠状动脉左前降支结扎40min,再灌注160min建立实验性心肌I/R损伤模型,于结扎冠状动脉10min后股静脉分别给于4、8、16mg/kg羟基红花黄色素A,测定心肌梗死面积、血清中乳酸脱氢酶(LDH)、血清肌钙蛋白(c Tn T)、肌酸激酶同工酶(CK-MB)活性的变化,血浆中血浆血栓素(TXB2)、6-酮-前列腺1a(6-keto-PGF1a)含量,心肌组织中IL-6、IL-1β、TNF-α水平和MPO活性。结果:羟基红花黄色素A 4、8、16mg/kg各剂量均可不同程度降低大鼠急性心肌梗死面积,减少心肌酶的漏出,升高6-keto-PGF1a含量,并可显著降低TXB2含量;羟基红花黄色素A 4、8、16mg/kg各剂量组可不同程度降低心肌组织炎性因子IL-6、IL-1β、TNF-α水平和MPO活性,其中对TNF-α水平和MPO活性的影响尤为显著。结论:羟基红花黄色素A对心肌I/R损伤具有较好的保护作用,可能与其抑制血小板聚集、抑制炎症反应有关。

羟基红花黄色素A注射剂对家兔血小板聚集、超微结构及血浆GMP-140含量的影响

目的:观察羟基红花黄色素A(HSYA)注射剂对家兔血小板聚集功能及超微结构的影响。方法:采用体内实验法,观察三种剂量的羟基红花黄色素A注射剂(10mg/kg、5mg/kg和2.5mg/kg)对由花生四烯酸(AA,0.35mmol/L)、二磷酸腺苷(ADP,300μmol/L)和血小板活化因子(PAF,3.6nmol/L)诱导的家兔血小板聚集作用的影响,以及血小板超微结构的变化;并单独研究了以PAF为诱导剂的情况下,血浆GMP-140的含量。结果:各剂量HSYA注射剂(10mg/kg、5mg/kg和2.5mg/kg)能抑制AA、PAF诱导的家兔血小板聚集;扫描电镜显示,各剂量HSYA注射剂能减少AA和PAF诱导后的聚集型血小板数量并使树突型血小板突起变少变短;另外,各剂量HSYA注射剂还能降低GMP-140的含量。结论:羟基红花黄色素A注射剂具有显著的抗PAF诱导的血小板聚集作用,抑制血小板的活化,从而为临床抗血小板聚集药物的使用提供更多选择。

羟基红花黄色素A可抑制大鼠血管平滑肌细胞增殖

目的 探讨羟基红花黄色素A（HSYA）对血管平滑肌细胞（VSMC）增殖的影响及其机制.方法 取清洁Ⅱ级雄性（Sprague Dawley,SD）大鼠腹主动脉,采用贴块法培养VSMC,DMEM培养基正常培养的VSMC为空白对照组、含有10 ng/ml PDGF培养基培养的VSMC为PDGF刺激组（PDGF组）、含有10 ng/ml PDGF培养基加入不同浓度的HSYA干预的VSMC为HSYA不同浓度组（HYSA 浓度分别为1、5、10、20、40及60 μmol/L）.采用MTT法分析不同浓度的HSYA对VSMC增殖的影响.采用蛋白印迹法检测各组增殖细胞核抗原（PCNA）蛋白表达水平及跨膜信号转导蛋白分子的磷酸化水平.采用免疫细胞化学法检测各组PCNA的表达.结果 HSYA不同浓度（1、5、10、20、40及60 μmol/L）组VSMC增殖水平均低于PDGF组（P均＜0.05）,且呈浓度依赖性.HYSA不同浓度（5、10、20及40 μmol/L）组PNCA蛋白表达水平均低于PDGF组（P均＜0.05）.PDGF组PDGFR磷酸化水平明显高于空白对照组（P＜0.05）,而HSYA不同浓度（5、10、20及40 μmol/L）组则均低于PDGF组,且呈浓度依赖性（P均＜ 0.05）.PDGF组MEK和ERK1/2磷酸化水平均明显高于空白对照组（P均＜0.05）,而HSYA不同浓度（5、10、20及40 μmol/L）组则均低于PDGF组,且呈浓度依赖性（P均＜0.05）.结论 HSYA可抑制VSMC增殖、降低PCNA表达,其可能是通过阻断MEK/ERK1/2信号通路抑制大鼠VSMC增殖的.