柄海鞘Styela clava化学成分的研究(Ⅱ)

从采自青岛海域的柄海鞘 Styela clava中分离得到两个化合物 (1)和 (2 )。经过 IR、MS、1HNMR和 13 CNMR等现代波谱学方法 ,确定二者的结构分别为 1- O-十六烷酰 -甘油酯和胸腺嘧啶。它们均为首次从该海域海鞘中得到。

柄海鞘(Styela clava)化学成分的研究

应用 GC- MS联用技术从柄海鞘 (Styela clava)中鉴定了 3个高级脂肪酸类化合物 :十六酸、十七酸、十八酸 ,以及六个甾醇类化合物 :(2 2 Z) - 2 6,2 7-二降 -麦角甾 - 5,2 2 -二烯 - 3β-醇、胆甾 - 5.2 2 -二烯 - 3β-醇、胆甾 - 8(1 4 ) -烯 - 3α-醇、豆甾 - 4 ,2 2 -二烯 - 3β-醇、(2 2 Z) - 5α-麦角甾 - 7,2 2 -二烯 - 3β-醇、豆甾 - 8(1 4 ) -烯 - 3β醇、豆甾 - 8(1 4 ) -烯 - 3β-醇。

柄海鞘Styela clava次生代谢产物的化学研究(Ⅰ)

从青岛产柄海鞘（Ｓｔｙｅｌａ ｃｌａｖａ）９５％乙醇提取物中分离得到６种次生代谢产物，经波谱（ＩＲ，ＭＳ，～１ＨＮＭＲ，～（１３）ＣＮＭＲ）分析，确定了它们的化学结构分别为异戊酰胺（ｉｓｏｖａｌｅｒａｍｉｄｅ）（１），胸腺嘧啶（ｔｈｙｍｉｎｅ）（２），胸腺嘧啶脱氧核苷（ｔｈｙｍｉｄｉｎｅ）（３），尿嘧啶（ｕｒａｃｉｌ）（４），尿嘧啶脱氧核苷（２′－ｄｅｏｘｙｕｒｉｄｉｎｅ）（５）和鲛肝醇（α－ｐａｌｍｉｔｙｌ ｇｌｙｃｅｒｉｎ ｅｔｈｅｒ）（６）。化合物１～６都系首次从柄海鞘Ｓｔｙｅｌａ ｃｌａｖａ中得到。

柄海鞘Styela clava中的甾醇化合物的分离和鉴定

应用GC-MS技术从青岛海域产柄海鞘Styelaclava中分离鉴定出5种甾醇化合物,分别是:(22Z)-26,27-二降-麦角甾-5,22-二烯-3β-醇(1),胆甾-5,22-二稀-3β-醇(2),胆甾醇(3),胆甾烷醇(4),(22E)-麦角甾-5,22-二烯-3β-醇(5)。

Observations on the morphology of embryonic and larval development in Styela canopus Savigny

The morphological characters on different developmental phases of embryo and larva and the schedule of the whole early developmental process in Styela canopus were observed and described. The types of reproduction and early development as well as the morphology, of egg and larva in different ascidian species were compared. Styela canopus is oviparous. Its egg, 230.4 similar to 336.0 mum in diameter, is equipped with extraembryonic cell layers measured 43.2 similar to 63.0 mum thick. The early development of Styela canopus is typical urodele development, including fertilization, cleavage, gastrulation, tadpole in membrane, tadpole, initiating metamorphosis and juvenile. The tadpole of Styela canopus, with a length of 0.6 similar to 0. 9 mm, consists of trunk and tail. There are obvious notochord, ocellus and adhesive papillae in the tadpole. Under the water temperature of (25 +/- 0-5) degreesC and the salinity of 27.0, the larva was hatched after 9.5 similar to 11.0 h since the fertilization.

Settlement and metamorphosis of Styela canopus Savigny larvae in response to some neurotransmitters and thyroxin

The larvae of ascidian Styela canopus Savigny were treated with epinephrine, norepinephrine, L-DOPA, GABA and thyroxin to test the ability of these compounds to induce or inhibit larval settlement and metamorphosis. The results showed that epinephrine, norepinephrine and L-DOPA at the concentration of 1 μmol/dm^3 induced larval settlement and metamorphosis in S. canopus, with short exposure （ 1 h） to 1 μmol/dm^3 of L-DOPA inducing rapid settlement. In contrast, GABA at the concentrations of 0.1 ～1130.0 μmol/dm^3 significantly inhibited the settlement and metamorphosis of S. canopus larvae. In addition, thyroxin at 1 -50 μg/dm^3 had no effect on larval settlement and metamorphosis in S. canopus. These results suggest the importance of neurotransmitters in the settlement and metamorphosis of S. canopus larvae.

The determination of larval metamorphic competence of Styela canopus Savigny

The larvae of Styela canopus show the highest rates of settlement and metamorphosis when treated with 40 mM KCl-elevated seawater at 25 degreesC in temperature and 27.0 in salinity. Induced by 40 mM KCl-elevated seawater, it took Styela canopus larvae approximately 3 h after hatching to develop competence to settle and metamorphose. The present experiment shows that Styela canopus larvae is an ideal model organism for examining the efficiency of antifouling substance.

Antiviral treatment of hepatitis B virus-transgenic mice by a marine organism, Styela plicata

瞄准：在肝炎 B 搬运人的一个鼠科的模型评估 Styela plicata 的有效成分的抗病毒的效果。方法：HBV 转基因的老鼠被划分成 3 个组(控制组， lamivudine 处理组和 Styela plicata 处理组的有效成分) 并且分配了收到正常饮食， lamivudine 或 Styela plicata 的有效成分连续星期。浆液肝炎 B 表面抗原被连接酶的免疫检测吸着剂试金(ELISA ) 方法。浆液 HBV DNA 被即时聚合酶链反应(RT-PCR ) 检测。浆液 T 助手(h) 1 cytokine interleukin (IL )-2 和 Th2 cytokine IL-6 被量的三明治酶免疫分析技术检测。另一组 HBV 转基因的老鼠被分配收到 Styela plicata 的有效成分连续星期。肝组织的组织学在治疗前后被评估。结果：12 个星期在开始治疗以后，浆液肝炎 B 表面抗原显著地在 Styela plicata 被降低与收到正常饮食的老鼠相比对待老鼠和对待 lamivudine 的老鼠(F (12wk )= 88.81， P (12wk )= 0.000 【0.01 ) 。浆液 HBV DNA 显著地在 Styela plicata 被降低与收到正常饮食的老鼠相比对待老鼠和对待 lamivudine 的老鼠(F (12wk )= 20.71， P (12wk )= 0.000 【0.01 ) 。然而， Styela plicata 的有效成分象 lamivudine 一样，不能完全禁止 HBV 的复制。在重量的单位的肝炎 B 表面抗原和 HBV DNA 的回缩现象一 could 在药的退却以后被发现 4 wk。八个星期在开始治疗以后，在 IL-2 的 Styela plicata 处理前后的浆液层次是 2.41 +/- 0.38 和 10.56 +/- 0.78 ng/L，分别地(t (8wk )=-16.51, P (8wk )= 0.000 【0.01 ) 。与在正常对待食谱的老鼠的 IL-2 的浆液层次相比(2.48+/-0.17 ng/L；t (8wk )= 13.23， P (8wk )= 0.000 【0.01 ) 。在 IL-6 的 Styela plicata 处理前后的浆液层次是 63.62 +/- 分别地， 6.22 ng/L 在正常对待食谱的老鼠 IL-6 与浆液相比铺平的 6.31 和 54.52 +/-(60.84 +/- 4.21 ng/L ) 。从 Styela 对待 plicata 的 HBV 转基因的老鼠的肝的组织学的分析也在发炎和肝炎 B 表面抗原显示出下降地位。结论：Styela plicata 可以是在对待长期的肝炎 B 的有效的抗病毒的药。

Therapeutic effect of Styela plicata on duck hepatitis B virus in vivo

Objective:To evaluate the antiviral activity of the alcohol extract of Styela plicata on DHBV (duck hepatitis B virus) in vivo. Methods: Guangzhou-Sheldrake ducklings congenitally infected with DHBV were assigned to receive the alcohol extract of Styela plicata or lamivudine for 30 consecutive days. The DHBV DNA of sera was detected by RT-PCR. and the histological analysis of duckling liver was evaluated. Results:Thirty days after therapy,histological analysis of duckling liver showed that the ducklings receiving the alcohol extract of Styela plicata or lamivudine exhibited catabatic status in the degree of liver cell degeneration and inflammation compared with the ducklings receiving normal diet. DHBV DNA of sera from alcohol extract of Styela plicata-treated ducklings and lamivudine-treated ducklings all produced significantly lower levels compared with ducklings receiving normal diet (P<0. 01 ). Although these treatment groups all exhibited a rebound phenomenon 10 d after withdrawal of medication, they still exhibited a significant lower level of serum DHBV DNA compared with the control group responded to normal diet (P<0. 05, P<0. 01). Conclusion:Styela plicata may be an effective antiviral medicine in treating chronic hepatitis B. The data of this experiment will be valuable in studying the therapeutic role and the potential therapeutic mechanism of Styela plicata.

Establishment of a developmental atlas and transgenetic tools in the ascidian Styela clava

The ascidian Styela clava is an ecologically important species that is distributed along coastal regions worldwide.It has a long history as a model animal for evolutionary and developmental biology research owing to its phylogenetic position between vertebrates and invertebrates,and its classical mosaic expression patterns.However,the standard developmental atlas and protocols and tools for molecular manipulation of this organism are inadequate.In this study,we established a standard developmental table and provided a web-based digital image resource for S.clava embryogenesis at each developmental stage from fertilized eggs to hatching larvae by utilizing confocal laser microscopy and 3D reconstruction images.It takes around 10 h for fertilized eggs to develop into swimming larvae and 20–30 min to complete the tail regression processes at the metamorphic stage.We observed that the notochord cells in S.clava embryos did not produce an extracellular lumen like Ciona robusta,but showed polarized elongation behaviors,providing us an ideal comparative model to study tissue morphogenesis.In addition,we established a chemical-washing procedure to remove the chorion easily from the fertilized eggs.Based on the dechorionation technique,we further realized transgenic manipulation by electroporation and successfully applied tissue-specific fluorescent labeling in S.clava embryos.Our work provides a standard imaging atlas and powerful genetic tools for investigating embryogenesis and evolution using S.clava as a model organism.

Correlativity between neural complex and gonadal development in Styela plicata

IT has been demonstrated that two reproductive hormones,gonadotropin-releasing hormone(GnRH)and gonadotropin(GTH),exist in the nervous system and Hatschek’s pit oflancelet,a species of Cephalochordata,and that these hormones are involved in the regulationof gonadal development and reproductive activity of lancelet.However,no report could

中国黄海柄海鞘的化学成分

目的:对采自中国黄海的柄海鞘(Styela clava)的化学成分进行研究,从中寻找有生物活性的次生代谢产物。方法:用硅胶柱层析和凝胶柱层析对柄海鞘的丙酮提取物进行分离纯化,根据其化学性质,结合现代波谱技术(MS,NMR等),对得到的化合物进行结构鉴定。结果:分离得到3个类胡萝卜素和1个芳香胺类化合物,其结构分别鉴定为mytiloxanthinone(1)、fucoxanthin(2)、all-trans-astaxanthin(3)和naphthalen-2-yl-phenyl-amine(4)。结论:这些化合物均系首次从柄海鞘中分离得到,并首次对化合物mytiloxanthinon(1)和fucoxanthin(2)的1H和13C NMR数据进行了全归属。

柄海鞘脂肪酸提取工艺的研究

用乙酸乙酯从柄海鞘(Styela clava)干粉中提取多不饱和脂肪酸,通过正交实验对提取工艺进行研究,得其最佳提取条件为:温度50℃,提取时间240 min,料液比1:10,提取次数2次。按此工艺,得脂肪酸产率为5.04%,利用气相色谱进行分析,得其中多不饱和脂肪酸质量分数为67.79%,EPA和DHA的质量分数之和为19.47%。

超临界CO_2提取海鞘脂肪酸的研究

用超临界CO2从柄海鞘(Styela clava)干粉中提取多不饱和脂肪酸。通过正交实验对提取工艺进行研究,得其最佳提取条件为:萃取温度50℃,萃取压力20 MPa,萃取时间2 h,夹带剂乙酸乙酯体积(mL):柄海鞘干粉质量(g)为1∶2。按此工艺,脂肪酸得率为19.5%。利用气相色谱进行分析,其多不饱和脂肪酸含量为75.77%,二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)含量之和为19.47%。

海绵和海鞘中可培养放线菌的分离与多样性比较

在相同的分离培养条件下,为比较地域差别较大的福建海域海绵动物(山海绵Mycale sp.和网架海绵Stylissa sp.)和海南海域海鞘动物(皱瘤海鞘Styela plicata和乳突皮海鞘Molgula manhattensis)之间可培养放线菌多样性的差异,作者采用5种放线菌分离培养基和1种细菌通用培养基,对海绵和海鞘中的放线菌进行分离培养。采用16S rRNA基因限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)分析和序列分析,揭示其多样性。共获得可培养放线菌198株,其中从海绵中分离到87株放线菌,从海鞘中分离到111株放线菌。RFLP分析表现为38种不同的图谱类型。16S rRNA基因序列分析表明,从海绵中分离到的放线菌包括6个放线菌属,其中有2株菌的16S rRNA基因序列与最相近的菌株相似性低于97%,可能是潜在的新菌株;从海鞘中分离到的放线菌包括7个放线菌属,有8株可能是潜在的新菌株。比较海绵和海鞘中可培养放线菌的多样性发现,从海绵中分离到的放线菌,除节细菌(Arthrobacter)以外,均包括在海鞘分离的放线菌属中。海鞘相关放线菌多样性水平不容忽视,是除海绵之外另一获得新型放线菌资源以及药用天然活性产物的重要来源。

皱瘤海鞘乙醇提取物抗乙肝HBsAg和HBeAg的初步研究

目的:研究皱瘤海鞘乙醇提取物的体外抗乙肝病毒作用。方法:用 2.2.15细胞株进行抗 HBsAg、HBeAg的试验研究。结果:海鞘乙醇提取物对 HBsAg、HBeAg的分泌均有较明显的抑制作用,对细胞的毒性低,其治疗指数均在10以上。结论:海鞘乙醇提取物体外具有明显的抗乙肝病毒作用。

柄海鞘共附生细菌的分离培养与系统发育多样性研究

通过纯培养和16SrRNA基因序列的相似性比对,并构建系统发育树对威海海域柄海鞘共附生细菌的多样性进行了研究。用2216E培养基从柄海鞘样品中分离到78株细菌,通过形态学特征和TCBS培养基选择性筛选后,选取30株具有代表性的菌株进行了16SrRNA基因测序和基于16SrRNA基因序列的系统发育多样性分析。结果表明,30株菌株分别属于细菌域的4个系统发育类群(Actinobacteria,Bacteroidetes,Firmicutes,Gro-teobacteria)的11个科,11个属。根据16SrRNA基因序列,大多数菌株与其系统发育关系最密切的模式菌株之间存在一定的遗传差异(16SrRNA基因序列相似性为91.82%～98.93%)。其中菌株HQW7,HQYD1,HQWD4,HQE1和HQE2与相关模式菌株的16SrRNA基因最高相似度仅分别为91.82%,92.51%,92.99%,93.52%,93.87%,这5株菌可能代表新的分类单元。威海海域柄海鞘共附生细菌存在着较为丰富的物种多样性和系统发育多样性,并可能存在新的物种。

海鞘脂肪含量及其脂肪酸组成

对烟台渤海湾的柄海鞘和玻璃海鞘的脂肪含量及其脂肪酸组成进行了分析,并对其提取工艺进行了研究。结果表明,海鞘中的脂肪以乙醚为溶剂,提取3h较为理想。脂肪总含量,玻璃海鞘为干重的6.16％;栖海鞘的含量为干重的5.02％,其中内囊的含量占10.68％、海鞘皮中的含量为1.14％。气相色谱及色—质谱分析的结果表明,柄海鞘内囊中的不饱和脂肪酸(UFA)占脂肪酸总量的 56.44％,其中多不饱和脂肪酸(PUFA)为37.57％,EPA+DHA占18.98％;玻璃海鞘中UFA占脂肪酸总量的36.68％,其中PUFA为23.34％,EPA+DHA占 9.8％。

柄海鞘化学成分研究

从采自青岛的柄海鞘 Styela clava中分离得到4个化合物,利用波谱学方法鉴定其结构分别为:△8,9-十八碳烯酸甘油酯(Ⅰ),胆甾-5,22-二稀-3β-醇(Ⅱ),胆甾醇(Ⅲ),胆甾-8,(14)-稀-3α-醇(Ⅳ)。其中化合物Ⅰ为首次从柄海鞘中分离得到。

冠瘤海鞘幼体附着和变态的离子控制

研究了人工海水中三种金属离子K+,Mg2+和Ca2+分别在不同浓度下对冠瘤海鞘(Styelaca nopusSavigny)幼体附着和变态的影响,同时采用K+通道阻滞剂TEA检验其可兴奋细胞膜上的K+通道特性,对冠瘤海鞘幼体附着和变态的离子控制机制进行初步探讨,结果表明:K+的浓度为20~40mmol/dm3 时诱导幼体附着和变态的效果最佳,低于4 5mmol/dm3 和高于80mmol/dm3 的K+浓度都表现出抑制效果;Mg2+浓度在0 ~ 32 mmol/dm3 时显著促进幼体附着和变态,达88mmol/dm3时起抑制作用;在人工海水中无论添加或减少Ca2+,均有效抑制幼体变态.TEA不能阻抑增加K+的诱导作用,这表明冠瘤海鞘幼体可兴奋细胞膜上的K+通道对外部TEA不敏感.另外,人工海水中缺乏K+或Ca2+或K+浓度达80mmol/dm3 都会对冠瘤海鞘幼体产生毒害作用.