苦参总碱抗柯萨奇B病毒作用机理的研究

在证实苦参具有抗柯萨奇Ｂ病毒（ＣＶＢ）作用的实验基础上，作者以纯化ＣＶＢ３作为病毒蛋白质对照，通过空斑测定及病毒蛋白质Ｌ－３５Ｓ－甲硫氨酸标记等方法，了解到纯化苦参总碱抗ＣＶＢ的作用机理是不影响病毒吸附、穿入，但它能进入细胞内影响病毒的生物合成，主要表现为抑制病毒蛋白质的合成。     

海南地不容总碱的镇痛抗炎作用及毒性观察

目的研究海南地不容总生物碱的镇痛抗炎作用及急性毒性。方法采用小鼠热板法、扭体法及甲醛测痛试验观察镇痛作用;采用二甲苯致小鼠耳肿胀、醋酸致小鼠腹腔毛细血管通透性、棉球诱导小鼠肉芽组织增生试验模型观察抗炎作用,通过胸腔白细胞游走试验研究其对炎症渗出液中白细胞数量的影响,应用交叉菜胶致足肿胀试验测定炎性组织中前列腺素E2（PGE2）含量并初步探讨其抗炎机理,急性毒性试验采用改良寇氏法测定LD50。结果海南地不容总碱能明显延长小鼠舔后足时间,减少扭体次数,降低甲醛测痛试验第一、二时相的疼痛强度;减轻小鼠耳肿胀,抑制毛细血管通透性增高及肉芽组织增生,有效抑制白细胞游走,并能明显减少大鼠炎症组织中PC-E2含量。其LD50为0．569g·kg-1,95％可信限为0．519～0．623g·kg-1。结论海南地不容总碱镇痛、抗炎作用显著,其机理可能与抑制PGE2的产生与释放有关。     

葛根芩连方药中黄连总生物碱的含量测定

目的 建立葛根芩连汤及其制剂中黄连总生物碱的含量测定方法。方法 氧化铝柱纯化,紫外分光光度法350 nm处测定含量。结果 葛根芩连汤水提物中总生物碱含量为32.4 mg/g,加样回收率为98.1% (RSD =2.9%)。葛根芩连微丸中总生物碱含量为19.6 mg/g。结论 该法测定黄连总生物碱方法简单、结果准确,可为葛根芩连微丸质量标准、葛根芩连方及其他含黄连的方剂中生物碱的含量测定提供参考。     

正交试验设计优选九节龙皂苷I聚乳酸微球的制备工艺

目的 筛选溶剂蒸发法制备九节龙皂苷I聚乳酸微球（ADS-I-PLA-MS）最佳工艺。方法 采用HPLC-ELSD测定方法,以包封率和载药量为评价指标,W/O/W溶剂蒸发法制备微球;通过单因素和正交试验设计,考察内水相九节龙皂苷I（ADS-I）甲醇溶液的质量浓度、ADS-I甲醇溶液与聚乳酸（PLA）二氯甲烷溶液体积比、PLA二氯甲烷溶液质量浓度和聚乙烯醇（PVA）体积等因素对ADS-I-PLA-MS包封率及载药量的影响。结果 溶剂蒸发法制备ADS-I-PLA-MS的最佳工艺条件为ADS-I甲醇溶液质量浓度为8 mg/mL、ADS-I甲醇溶液与PLA二氯甲烷溶液体积比为1∶13、PLA二氯甲烷溶液质量浓度为90 mg/mL、PVA体积为500 mL。结论 优选出的ADS-I-PLA-MS制备工艺合理可行。     

蝙蝠葛酚性碱与褪黑素对缺血再灌注损伤神经元的作用比较

BACKGROUND： Recent studies have demonstrated that phenolic alkaloids from Menispermum dauricum （PAMD） can protect the heart and brain from ischemia/reperfusion injury, and promote neuron survival by inhibiting neuronal Bax and upregulating Bcl-2 expression following ischemia/reperfusion.
OBJECTIVE： To investigate the neuroprotective effects of PAMD versus exogenous melatonin against ischemia/reperfusion injury.
DESIGN, TIME AND SETTING： Observation and comparison experiments at a cellular level were performed at the Department of Biochemistry and Molecular Biology, Tongji Medical College of Huazhong University of Science and Technology between February 2007 and February 2008.
MATERIALS： PAMD （95% purity） was provided by Kunming Institute of Botany, Chinese Academy of Sciences; melatonin was provided by Sigma, USA.
METHODS： N2a mouse neuroblastoma cells were cultured in vitro deprived of glucose, serum and oxygen for 90 minutes, then cultured in normal medium containing different concentrations of PAMD （0.1, 1.0, 10 mg/L） or melatonin （1, 10, and 100 μmol/L）. Cells cultured in normal conditions served as a control.
MAIN OUTCOME MEASURES： The culture solution was collected to determine the content of ex- citatory neurotransmitters such as glutamic acid and aspartic acid; cell viability was detected by MTT methods; reactive oxygen species production was determined by fluorescence spectroscopy; mito- chondrial transmembrane potential （？Ψm） was detected by laser confocal scanning; cytochrome C was measured by western blotting; and caspase-3 activity was determined by visible spectropho- tometry.
RESULTS： Melatonin and PAMD both promoted oxygen-glucose-serum deprivation-mediated N2a cell survival （P 〈 0.01） and inhibited glutamic acid release （P 〈 0.01）, but melatonin did not inhibit aspartic acid production. The protective effects were the strongest using melatonin 100 μmol/L and PAMD 10 mg/L, so subsequent experiments were the performed at those doses. Although PAMD could no longer maintain mitochondrial transmembrane potential 6 hours after reperfusion, its in- hibitory effects on cytochrome C release from mitochondria and scavengers of reactive oxygen species were stronger than those of melatonin （P 〈 0.01）. However, its inhibitory effect on caspase-3 activity was weaker than that of melatonin： PAMD could inhibit caspase-3 activity 12 hours after reperfusion （P 〈 0.01）, but melatonin inhibited caspase-3 activity 28 hours after reperfusion （P 〈 0.01）.
CONCLUSION： The results show that melatonin and PAMD have neuroprotective effects, but that the mechanisms are varied. Melatonin can maintain mitochondrial transmembrane potential, but its inhibitory effects on cytochrome C release, caspase-3 activity, and reactive oxygen species scav-enging are different from those of PAMD.     

阳离子交换树脂酸水洗脱法纯化苦参总碱的工艺优选

目的优选阳离子交换树脂不同浓度盐酸溶液洗脱纯化苦参总碱的最佳工艺条件。方法以苦参碱、氧化苦参碱和总生物碱的洗脱率为考察指标,以盐酸溶液作为洗脱剂,优选阳离子交换树脂纯化苦参总碱的最佳工艺和参数条件。结果苦参总碱阳离子交换树脂纯化洗脱条件为:以5%盐酸溶液作为洗脱剂,以6 BV.h-1的流速洗脱,苦参碱和氧化苦参碱、苦参总碱洗脱率均可达85%。结论采用阳离子交换树脂酸水洗脱法纯化苦参总碱效率高,工艺简便,可用于苦参总碱的提取纯化。     

毛苞瓜叶乌头生物碱成分的研究

目的 研究毛苞瓜叶乌头根中的生物碱化学成分.方法 酸提碱沉提取总碱,硅胶层析法分离,波谱法鉴定结构.结果 分离到6个已知的去用二萜生物碱,分别为滇乌碱(yunaconitine)、塔拉萨敏(talasamine)、瓜乌定(hemsleyadine)、拳乌宁乙(circinanine B)、黄草乌碱丁(schaconitine)和senbusine A.结论 应用光谱法和与已知物TLC对照的方法鉴定了所有化合物的结构.     

HPLC法测定苦参总碱中苦参碱与氧化苦参碱的含量

目的:采用高效液相色谱法测定苦参总碱中苦参碱和氧化苦参碱的含量。方法:采用 Elite Hypersil NH_2柱(5μm,250mm×4.6 mm),乙腈-无水乙醇-3%磷酸溶液(81:10:9)为流动相,流速:1.0 mL·min~(-1),检测波长220 nm,柱温:25℃,进样量:10 μL。结果:苦参碱、氧化苦参碱的线性范围分别为5.32～53.2μg·mL~(-1)(r=0.9997),65.7～525.6μg·mL~(-1)(r=0.9995)。苦参碱平均加样回收率为99.5%,RSD=1.0%(n=6);氧化苦参碱的平均加样回收率为99.7%,RSD=0.6%(n=6)。结论:本方法操作简便、快速、可靠,可用于控制苦参总碱质量。     

Study on inhibitory effect by total alkaloids in Capparis spinosa on SGC-7901 in vitro

Objective To investigate the apoptosis effect of Total alkaloids on human gastric cancer cells SGC-7901 from Capparis spinosa(C.S)and possible mechanism of it.Methods SGC-7901 cells were treated with different concentrations of the Total alkaloids in CS.MTT assay and SRB assay were used to observe the inhibitory rate of the Total alkaloids,and fluorescence microscope,flow cytometry and used to observe the influence of the Total alkaloids on cell apoptosis and cell cycle changes of SGC-7901.Results The results showed total alkaloids can inhibit the growth of human gastric adenoma cells SGC-7901.Measurements using mononuclear cell direct cytotoxicity assay(the MTT method)shows that its cytotoxic effect on SGC-7901 is strong,with IC50 being 142.895 μg·mL-1,respectively.Results from SRB assay show that the anticancer effect of Total alkaloids is cytostatic at low concentration,with LC50 for this cells being 41.271 μg·mL-1,respectively,but it becomes mainly cytotoxic at high concentration,with GI50 for SGC-7901 being 244·932 μg·mL-1,respectively.Total alkaloids can induce apoptosis in tumor cells.Forty-eight hours after they are treated with total alkaloids of different concentrations,SGC-7901 cells are stained with Hoechst33258 fluorochromes.Observation using a fluorescence microscope reveals that total alkaloids can cause the chromatin in tumor cell nuclei to condense and fragment.The nuclei condense into a uniform,dense mass and then break up.Sprouts keep on forming on the cell membrane and then dropping off,so that the cell breaks up into several apoptotic bodies of different sizes.As total alkaloids concentration is increased,these morphological changes under the microscope become more and more clear,indicating that the proportion of cells undergoing apoptosis is gradually increasing.After treating with 75,150 and 300 μg·mL-1 of the total alkaloids in C.S for 72 h,the apoptotic rates of SGC-7901 cells were 8.7%,14.309%,0.819%.Conclusions Inducing apoptosis is one of the anti-cancer mechanism of total alkaloids of C.S.