枸杞多糖的分离鉴定与体外肠道代谢初步分析

对枸杞多糖的分离纯化工艺和结构鉴定、质量分析方法进行研究,并对其体外肠道代谢过程进行模拟分析,为其开发利用提供参考。枸杞子经乙醇提取除杂、水提、超滤浓缩、冷冻干燥获得枸杞多糖(LBP);LBP采用中压制备色谱系统在线检测,先经DEAE-650M阴离子交换色谱柱分离获得4个主要组分,再分别经超滤浓缩、Sephadex G25色谱纯化、冷冻干燥,获得纯化的4种主要枸杞多糖LBP-1～4。经HPSEC色谱分析,组分LBP-1～4相对分子量依次为65.58、63.83、62.67、65.58 kD;采用HPAEC-PAD检测,表明LBP-1～4主要由岩藻糖、鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖等6种单糖组成,其中阿拉伯糖与半乳糖含量较高;采用紫外-可见分光光度法分析,测定LBP-1～4的中性糖、糖醛酸和总蛋白质含量分别为64.08%～73.05%、8.5%～18.0%、7.27%～18.67%;体外模拟肠道代谢分析结果表明,LBP在人工胃液中稳定,在人工肠液、大鼠肠道菌群液中会发生降解代谢。上述研究结果可为枸杞多糖的制备与质量控制、产品应用开发提供参考。     

小球藻中类胡萝卜素类化合物的高效液相色谱分离条件的优化

采用超声破碎法和甲醇:二氯乙烷=2:1(v/v)溶剂来萃取小球藻中类胡萝卜素类化合物.分别采用了3种不同的色谱分离方法,最后确定小球藻中类胡萝卜素类化合物的最优色谱分离条件为甲醇(A):乙氰(B)=90:10(v/v),柱温为室温,流速1ml/min.用ZOBAX SB-C18色谱柱和ZOBAX Eclipse plus C18色谱柱进行分离试验时发现,ZOBAX Eclipse plus C18色谱柱的分离效果良好.最后确定小球藻中叶黄素含量为2.312mg/g.     

肉色链孢囊菌和桔橙指孢囊菌发酵液中化学成分的研究

采用硅胶柱色谱、凝胶柱色谱等分离手段,利用波谱分析方法,分别对放线菌肉色链孢囊菌(Streptospor-angium carneum)与桔橙指孢囊菌(Dactylosporangium aurantiacum)发酵液中的化合物进行分离鉴定。从肉色链孢囊菌发酵液中分离得到3个化合物,分别鉴定为腺嘌呤核苷(1),2′-脱氧腺苷(2),5,7,4′-三羟基异黄酮(3)。从桔橙指孢囊菌发酵液中也分离得到3个化合物,分别鉴定为腺嘌呤核苷(1),5,7,4′-三羟基异黄酮(3),2′-脱氧尿嘧啶核苷(4)。     

金针菇固体发酵菌丝体次级代谢产物分离鉴定

采用硅胶柱色谱、ODS柱色谱、Sephadex LH-20柱色谱、HPLC等分离方法对金针菇大米发酵乙酸乙酯提取物进行分离,根据理化性质和波谱数据鉴定化合物结构：鉴定了4个倍半萜,包括1个新的桉叶烷型倍半萜,3个侧柏烷型倍半萜,分别是flamvelutpenol A(1),aquaticol(2),enokipodin C(3),limacellone(4)。并通过与Rh2(OCOCF3)4络合的方法确定了新化合物flamvelutpenol A(1)和limacellone(4)的绝对构型。其中化合物3具有较好的抗菌活性,对耐甲氧西林金黄色葡萄球菌和枯草芽孢杆菌的MIC值分别为12.5mg/L,25mg/L。且化合物1－4均是首次从该种真菌中分离得到。     

黄芪多糖与枸杞多糖联用对小鼠肝组织损伤的保护作用

探究黄芪多糖与枸杞多糖对四氯化碳(CCl_4)所致小鼠急性肝损伤的协同保护作用。采用CCl_4诱导小鼠急性肝损伤,动物处死后取血液测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)活性,取肝脏计算肝指数并制备肝匀浆测定其中谷胱甘肽(GSH)活力、丙二醛(MDA)含量,并对小鼠肝脏进行组织切片观察,评价黄芪多糖与枸杞多糖联用对小鼠肝组织的保护效果。实验显示,黄芪多糖与枸杞多糖各配比(LBP 50 mg/kg+APS 100,LBP 70 mg/kg+APS 400,LBP 350 mg/kg+APS 800)均能显著抑制CCl_4急性肝损伤所引起的MDA含量、肝脏指数、ALT和AST活性的升高(p0.05),有效地提高肝脏中GSH的含量,减轻小鼠的肝组织损伤程度。本研究表明黄芪多糖与枸杞多糖联用对CCl_4诱导的小鼠急性肝损伤有明显的保护作用。     

玄参化学成分研究（英文）

采用硅胶柱色谱、反相柱色谱、凝胶LH-20柱色谱、制备液相色谱等方法从中药玄参中分离得到15个化合物,并结合波谱数据及理化性质对其进行结构鉴定,分别鉴定为:哈巴俄苷(1)、哈巴苷(2)、益母草苷A(3)、6-O-甲基梓醇(4)、京尼平苷(5)、6-O-α-L-鼠李糖基桃叶珊瑚苷(6)、ningpogenin(7),甘草素(8),邻苯二甲酸二(2-乙基己)酯(9),麦芽糖(10),熊果酸(11),丙三醇(12),5-羟甲基糠醛(13),胡萝卜苷(14)及β-谷甾醇(15)。其中化合物3、6、8、9、10、12均为首次从该植物中分离得到。     

深海放线菌Actinomadura sp.01119的代谢产物研究

采用硅胶柱色谱、制备薄层色谱和凝胶色谱等分离方法,对分离自深海沉积物中的海洋放线菌Actinomadura sp.01119的代谢产物进行研究,从中分离得到9个化合物,经1H NMR、13C NMR和MS等波谱分析并结合相关文献鉴定为环(L-苯丙-L-脯)二肽(1),环(L-脯-L-脯)二肽(2),环(L-酪-L-脯)二肽(3),环(D-酪-L-脯)二肽(4),环(L-异亮-L-脯)二肽(5),环(L-缬-L-脯)二肽(6),棕榈酸(7),对羟基苯甲酸(8),尿嘧啶(9)。其中化合物1~6为环二肽类化合物。     

高速逆流色谱制备玫瑰红景天中的三种酚性化合物

采用高速逆流色谱方法(HSCCC,High-speed Counter-current Chromatography)同时分离三种玫瑰红景天酚性化合物。玫瑰红景天提取物经聚酰胺吸附多酚后经硅胶柱分级得预分离样品,采用正己烷∶乙酸乙酯∶甲醇∶水(4∶5∶4∶5,v/v/v/v)组成的两相溶剂系统对预分离样品进行分离纯化,一次进样150 mg,一次色谱分离得到化合物1:68.5 mg、化合物2:8.5 mg、化合物3:45.5 mg,纯度都超过98%。通过ESI-MS、1H NMR对其结构进行鉴定化合物1为没食子酸(Gallic acid),化合物2为没食子酸甲酯(Methyl gallate),化合物3为山奈酚(Kaempferol)。结果表明利用HSCCC可以成功分离三种酚性化合物,分离效果好,产品纯度高。     

小蓬草精油的分离组分比较分析

通过薄层色谱分析和柱色谱分离结合气相色谱-质谱分析小蓬草精油的分离组分,结果表明其不同分离组分组成有差异,可以分离得到以香芹酮(18.33%)为特征成分,含有cis-香芹醇(3.79%),cis-香芹酮氧化物(1.74%),trans-香芹醇(1.62%),橙花叔醇(1.13%),石竹烯氧化物(0.38%),对甲基苯乙酮(0.64%)或者是以cis-香芹醇为特征成分,含有cis-马鞭草烯酮的分离组分。香芹酮、cis-香芹醇、橙花叔醇、对甲基苯乙酮、cis-马鞭草烯酮等是具有香气的化合物。     

海芒果叶中三萜类成分的研究

采用多种柱色谱等分离纯化手段对海芒果叶中的三萜类成分进行分离,得到7个化合物,通过理化性质和波谱方法鉴定化合物为熊果醇(1),(23Z)-9,19-cycloart-25-ene-3β,24-diol(2),大戟醇(3),乌苏酸(4),2α-羟基乌苏酸(5),乙酰乌苏酸(6),α-香树脂醇(7)。化合物1～6均为首次从该属植物中分离得到。     

Hypoglycemic and hypolipidemic effects and antioxidant activity of fruit extracts from Lycium barbarum

The hypoglycemic and hypolipidemic effects of Lycium barbarum fruit water decoction, crude polysaccharide extracts (crude LBP), and purified polysaccharide fractions (LBP-X) in alloxan-induced diabetic or hyperlipidemic rabbits were investigated through designed sequential trials and by measuring blood glucose and serum lipid parameters. Total antioxidant capacity was also assessed using trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assay. It was found that the three Lycium barbarum fruit extracts/fractions could significantly reduce blood glucose levels and serum total cholesterol (TC) and triglyceride (TG) concentrations and at same time markedly increase high density lipoprotein cholesterol (HDL-c) levels after 10 days treatment in tested rabbits, indicating that there were substantial hypoglycemic and hypolipidemic effects. Hypoglycemic effect of LBP-X was more significant than those of water decoction and crude LBP, but its hypolipidemic effect seemed to be weaker. Total antioxidant capacity assay showed that all three Lycium barbarum extracts/fractions possessed antioxidant activity. However, water and methanolc fruit extracts and crude polysaccharide extracts exhibited stronger antioxidant activity than purified polysaccharide fractions because crude extracts were identified to be rich in antioxidants (e.g., carotenoids, riboflavin, ascorbic acid, thiamine, nicotinic acid). Lycium barbarum polysaccharides (glycocojugates), containing several monosaccharides and 17 amino acids, were major bioactive constituents of hypoglycemic effect. Both polysaccharides and vitamin antioxidants from Lycium barbarum fruits were possible active principles of hypolipidemic effect.     

枸杞多糖对H5N1流感全病毒灭活疫苗的体液免疫增强作用

探讨枸杞多糖(Lycium barbarum polysaccharide,LBP)作为佐剂对H5亚型流感病毒全病毒灭活疫苗的体液免疫增强效果。将流感病毒A/Vietnam/1194/2004(H5N1)灭活疫苗与不同剂量的枸杞多糖混合后以腹腔注射的方式共同免疫小鼠,免疫后三周收集血清用于特异性抗体检测。实验中设立氢氧化铝佐剂组作对照共同评价LBP作为佐剂的免疫增强效果。结果显示,小鼠血清中针对H5灭活疫苗的特异性抗体水平在一定范围内随着LBP剂量的增加而提高。LBP在800μg剂量时血清特异性抗体水平较无佐剂组显著增强,并与氢氧化铝佐剂组大致相当。因而,LBP有可能成为一种有效的流感灭活疫苗免疫佐剂。     

Effect of lycium barbarum polysaccharide on human hepatoma QGY7703 cells: inhibition of proliferation and induction of apoptosis

Lycium barbarum polysaccharide (LBP), extracted from Lycium barbarum that is a kind of traditional Chinese herb, is found to have anticancer activity. In this study, the effect of LBP on the proliferation rate, cell cycle distribution and apoptosis in the human hepatoma QGY7703 cell line were investigated. The effects of this compound were also tested on the concentration of calcium in cells. LBP treatment caused inhibition of QGY7703 cell growth with cycle arrest in S phase and apoptosis induction. The amount of RNA in cells and the concentration of intracellular Ca2+ were increased. Moreover, the distribution of calcium in cells was changed. Taken together, the study suggests that the induction of cell cycle arrest and the increase of intracellular calcium in apoptotic system may participate in the antiproliferative activity of LBP in QGY7703 cells.     

Polysaccharides from wolfberry prevents corticosterone-induced inhibition of sexual behavior and increases neurogenesis

Lycium barbarum, commonly known as wolfberry, has been used as a traditional Chinese medicine for the treatment of infertility and sexual dysfunction. However, there is still a scarcity of experimental evidence to support the pro-sexual effect of wolfberry. The aim of this study is to determine the effect of Lycium barbarum polysaccharides (LBP) on male sexual behavior of rats. Here we report that oral feeding of LBP for 21 days significantly improved the male copulatory performance including increase of copulatory efficiency, increase of ejaculation frequency and shortening of ejaculation latency. Furthermore, sexual inhibition caused by chronic corticosterone was prevented by LBP. Simultaneously, corticosterone suppressed neurogenesis in subventricular zone and hippocampus in adult rats, which could be reversed by LBP. The neurogenic effect of LBP was also shown in vitro. Significant correlation was found between neurogenesis and sexual performance, suggesting that the newborn neurons are associated with reproductive successfulness. Blocking neurogenesis in male rats abolished the pro-sexual effect of LBP. Taken together, these results demonstrate the pro-sexual effect of LBP on normal and sexually-inhibited rats, and LBP may modulate sexual behavior by regulating neurogenesis.     

Protective effect of Lycium barbarum polysaccharides on oxidative damage in skeletal muscle of exhaustive exercise rats

The aim of this study was to determine the modulatory effect of Lycium barbarum polysaccharides (LBP) on the oxidative stress induced by an exhaustive exercise. 32 male Wistar rats were taken in the study. The experiment was a 30-day exhaustive exercise program. We determined the lipid peroxidation, glycogen levels, and anti-oxidant enzyme activities in skeletal muscle. The results demonstrated that L. barbarum polysaccharides administration significantly increases glycogen level and anti-oxidant enzyme activities, and decreased malondialdehyde (MDA) level and creatine kinase activities. In conclusion, L. barbarum polysaccharides administration can significantly decrease the oxidative stress induced by the exhaustive exercise.     

Lycium barbarum polysaccharides: Protective effects against heat-induced damage of rat testes and H2O2-induced DNA damage in mouse testicular cells and beneficial effect on sexual behavior and reproductive function of hemicastrated rats

Lycium barbarum, a famous Chinese medicinal herb, has a long history of use as a traditional remedy for male infertility. Polysaccharides are the most important functional constituent in L. barbarum fruits. We systematically investigated the effect of L. barbarum polysaccharides (LBP) on rat testis damage induced by a physical factor (43 degrees C heat exposure), on DNA damage of mouse testicular cells induced by a chemical factor (H2O2), and on sexual behavior and reproductive function of hemicastrated male rats. The results showed that LBP provided a protective effect against the testicular tissue damage induced by heat exposure. When compared with negative control, a suitable concentration of LBP significantly increased testis and epididymis weights, improved superoxide dismutase (SOD) activity, and raised sexual hormone levels in the damaged rat testes. LBP had a dose-dependent protective effect against DNA oxidative damage of mouse testicular cells induced by H2O2. LBP improved the copulatory performance and reproductive function of hemicastrated male rats, such as shortened penis erection latency and mount latency, regulated secretion of sexual hormones and increased hormone levels, raised accessory sexual organ weights, and improved sperm quantity and quality. The present findings support the folk reputation of L. barbarum fruits as an aphrodisiac and fertility-facilitating agent, and provide scientific evidence for a basis for the extensive use of L. barbarum fruits as a traditional remedy for male infertility in China.     

枸杞色素及多糖的不同提取次序对其得率及抗氧化活性的影响

为探究先后提取枸杞多糖及枸杞色素时对各自得率的影响。本研究通过分别考察这两种成分在提取过程中的提取溶剂、提取温度、料液比、提取次数及提取时间等因素对枸杞色素和枸杞多糖的得率影响,确定枸杞色素和枸杞多糖在提取次序不同时,两者的最佳提取工艺以及对DPPH·和·OH自由基清除率。结果表明,首先提取枸杞多糖后,枸杞色素的最佳提取工艺为采用正己烷,80℃时,料液比1∶10,提取2次,每次1 h,枸杞色素得率为2.48%,此时枸杞多糖得率为7.45%;而首先提取枸杞色素后,采用了超声辅助提取的方式提取枸杞多糖,发现超声效率为25%,料液比1∶10,提取20 min,枸杞多糖得率为5.23%,此时枸杞色素得率为3.93%。因此,首先提取枸杞多糖,使其平均得率为7.45%,而后提取枸杞色素,其平均得率为2.48%;总体上,枸杞色素1和枸杞多糖1对DPPH·自由基清除率都较高,枸杞多糖1对·OH自由基清除率较高,其抗氧化活性都接近Vc。     

转基因枸杞中蛋白酶体糜蛋白酶样活性的研究

利用提取的转基因枸杞和正常枸杞蛋白与蛋白酶体的特异性荧光底物Suc-LLVY-AMC室温孵育,于360nm（激发光）/460nm（发射光）波长下测定荧光值的方法,研究了转基因枸杞和正常枸杞中蛋白酶体糜蛋白酶活性的差异及其特异性抑制剂MG115对枸杞中蛋白酶体活性的影响。结果表明表达H IV壳体蛋白的转基因枸杞蛋白酶体活性是空载体对照转基因枸杞蛋白酶体活性的3.6倍,是正常枸杞的4.2倍。10μM的MG115对表达H IV壳体蛋白的转基因枸杞蛋白酶体活性抑制率为87%,携带空载体对照的转基因枸杞抑制率为74%,而对正常枸杞抑制率仅为8.6%;50μM的MG115抑制作用与10μM的相比没有明显变化。这一结果将为利用蛋白酶体抑制剂提高转基因枸杞悬浮细胞H IV-1CA含量的研究提供理论依据。     

枸杞多糖与黄芪多糖抑菌活性的研究*

目的:研究黄芪多糖和枸杞多糖的抑菌活性并探讨不同pH值对其抑菌活性的影响。方法:采用滤纸片扩散法,分析不同浓度黄芪多糖和枸杞多糖在不同pH值下对几种常见细茵和霉菌(大肠杆菌、沙门氏菌、金黄色葡萄球菌、黑曲霉、产黄青霉)的抑制效果。结果:对于细菌,枸杞多糖8mg/mL时出现抑菌圈,而黄芪多糖0.02 mg/mL时效果最佳;对于霉菌,随着枸杞多糖浓度的增大,抑菌圈的直径增大,而黄芪多糖0.02 mg/mL时效果最佳;当枸杞多糖和黄芪多糖在pH6的条件下,二者抑菌活性均最强。结论:枸杞多糖和黄芪多糖对细菌、霉菌都有一定的抑制效果,pH值可影响枸杞多糖和黄芪多糖的抑菌效果。     

不同地域宁夏枸杞活性成分的研究

对不同地域宁夏枸杞中活性成分的含量进行了分析。研究结果显示产地不同的宁夏枸杞中活性成分含量有明显差别,宁夏银川产的宁夏枸杞中所含的活性成分高于辽宁朝阳产的宁夏枸杞;在积温高、土壤呈弱碱性条件下,宁夏枸杞中的多糖、游离氨基酸、果红素含量较高。