总状蕨藻盾叶变种寡糖的酶解制备及其免疫调节活性的作用机制

将总状蕨藻盾叶变种多糖经乳糖酶酶解、膜过滤、透析和冻干制备获得总状蕨藻盾叶变种寡糖(Caulerpa racemosa var.peltate oligosaccharide,CRVO-G),测定CRVO-G的相对分子质量与单糖组成,评价CRVO-G的免疫调节活性,并对其作用机制展开研究。结果显示,CRVO-G主要是由相对分子质量分别为617.115、621.081和995.368的2种四糖和1种六糖组成的寡糖混合物,主要由半乳糖(51.0%)、甘露糖(21.3%)、葡萄糖(11.5%)、木糖(6.5%)、鼠李糖(2.3%)、葡萄糖醛酸(2.0%)、盐酸氨基葡萄糖(1.8%)、岩藻糖(0.8%)、盐酸氨基半乳糖(0.7%)、半乳糖醛酸(0.7%)、甘露糖醛酸(0.7%)和阿拉伯糖(0.6%)组成。此外,CRVO-G具有良好的免疫调节活性,能促进一氧化氮(NO)和免疫因子的释放,如白细胞介素-1β、白细胞介素-6和肿瘤坏死因子-α。代谢组学和蛋白免疫印迹结果表明,CRVO-G可以促进巨噬细胞花生四烯酸代谢通路中血栓素A2和15-酮前列腺素F2α的合成,提高环氧合酶-2(cyclooxygenase-2,COX-2)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和核因子κB(nuclear factorκB,NF-κB)的磷酸化蛋白的表达水平。研究表明,在巨噬细胞RAW264.7细胞中CRVO-G通过NF-κB/iNOS/COX-2途径发挥免疫调节作用。

总状蕨藻多糖提取工艺的研究

采用酶法提取与热水抽提相结合的方法研究了总状蕨藻多糖的提取工艺,结果显示,采用固液质量比1∶10,蛋白酶用量136.5 U/g,于60℃下酶法提取2.5 h后,再将固液质量比调至1∶20,在温度95℃条件下继续进行热水提取2.5 h,多糖得率为6.7%,比单用热水提取法提高了131.0%。

总状蕨藻水溶性成分的提取及其对羟自由基清除作用

以清除Fenton反应产生的羟自由基能力为考察指标,采用单因素实验和正交优化实验研究了总状蕨藻水溶性成分的提取工艺;以抗坏血酸(Vc)作阳性对照,测定了不同浓度水溶性成分对羟自由基(.OH)的清除作用。结果表明,在温度60℃、料液比1:25、时间1 h的条件下提取的水溶性成分清除羟自由基的能力最强,在此工艺条件下,水溶性成分的得率为3.09%。当浓度为0.187 g/L^0.75 g/L时,总状蕨藻水溶性成分对羟自由基清除能力较VC强,且存在剂量-效应关系,最大清除率为72.35%。总状蕨藻水溶性成分和VC的IC50分别为0.513 g/L和0.665 g/L。

总状蕨藻多糖体外抗肿瘤作用

以总状蕨藻为原料,采用热水与蛋白酶酶解相结合浸提得到总状蕨藻多糖提取液,经Sevag法和透析法等步骤纯化后得到总状蕨藻粗多糖(Caulerpa racemosa polysaccharide,CRP)。紫外扫描结果表明CRP中含有少量蛋白质。红外光谱分析CRP有一般糖类物质的特征吸收峰,含硫酸基团,是一种酸性多糖。采用噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)法研究了CRP对HeLa、K562、CNE-2z细胞的抑制作用,结果表明CRP对3种肿瘤细胞都有一定的抑制作用,在24~72 h内,随着时间的延长,抑制率逐渐增强,在0.062 5~4 mg/mL质量浓度范围内,随着质量浓度的增加,抑制率逐渐增强,对HeLa、K562和CNE-2z细胞的抑制率最大分别达到61.7%、85.9%和90.3%。

总状蕨藻盾叶变种多糖对小鼠T细胞亚群及NK细胞免疫功能的调节

目的:观察总状蕨藻盾叶变种多糖对正常小鼠T细胞亚群及NK细胞的免疫调节作用。方法:腹腔注射总状蕨藻盾叶变种多糖溶液,用流式细胞仪检测血中免疫参数(CD3+,CD4+,CD8+T细胞亚群百分比)和脾脏NK细胞百分比,并测定胸腺、脾脏指数。结果:总状蕨藻盾叶变种多糖能明显降低小鼠血中CD3+,CD4+,CD8+百分比(P<0.05),同时增强脾脏NK细胞百分比(P<0.05),并显著降低胸腺指数(P<0.05),增加脾脏指数(P<0.05)。结论:总状蕨藻盾叶变种多糖对小鼠T细胞亚群和NK细胞具有双向的免疫调节作用。

超声波法提取总状厥藻中硫酸酯多糖的工艺研究

研究了超声波法从总状厥藻(Caulerpa racemosa)中提取厥藻硫酸酯多糖的工艺。考察了溶剂体积与藻粉质量配比(液固比)、超声提取时间、超声波功率对多糖提取率的影响。在室温下,液固比为20∶1,800 W超声功率下超声处理30 min能得到最佳的多糖提取率。采用优化工艺,多糖得率可达4.4%,比传统水提法提高了63.6%,色素等杂质的析出也减少,是一种良好的提取硫酸酯多糖的方法。

UPLC-MS/MS测定地表水中总微囊藻毒素的快速前处理方法探究

为方便地表水中总微囊藻毒素(TMCs)的预警监测,探究提取MC-LR、MC-RR、MC-YR的不同快速前处理方法,建立同时测定3种TMCs的煮沸-过滤-UPLC-MS/MS法。该方法在0.006μg/L~50.0μg/L范围内线性良好,方法检出限为0.006μg/L~0.010μg/L,实际水样3个质量浓度水平的加标回收率为88.6%~108%,6次测定结果的RSD为2.6%~9.4%。

超声共振技术在水库藻类抑制中的应用初探

以深圳某水库为例,初步探索了超声共振技术对藻类的抑制效果。研究结果表明,该水库藻密度达到了107个/L以上,超过了饮用水源中藻类危险限值,且藻类浓度随温度升高而升高。在水库中设置的围栏(约1 600m2)并安装超声共振设备(功率5 W),总藻浓度降低了25.4%与30.4%,蓝藻降低了39.9%与23.5%。超声共振技术为控制水库藻类提供了潜在的技术选择。

总状蕨藻盾叶变种多糖及其硒化产物对小鼠T细胞和NK细胞的调节

一种分离自总状蕨藻盾叶变种的多糖CrvpPS与纳米硒反应之后形成稳定的多糖纳米硒(CrvpPS-nano-Se)系统.实验检测了CrvpPS和CrvpPS-nano-Se对小鼠T淋巴细胞亚群及NK细胞的调节功能.用氢化可的松制造免疫功能低下的小鼠动物模型,给模型小鼠分别连续10天灌胃CrvpPS、蔗糖-纳米硒、CrvpPS-nano-Se溶液和阳性对照药物左旋咪唑,检测脾脏和胸腺指数的变化,并用流式细胞仪检测血中T细胞亚群(CD3+,CD4+,CD8+)和脾脏NK细胞百分比.结果表明,CrvpPS和CrvpPS-nano-Se对胸腺有较大的刺激作用,能使模型小鼠的CD3+,CD3+CD4+,NK细胞百分比以及CD4+/CD8+明显升高(P<0.05),CrvpPS-nano-Se提升CD3+,CD3+CD4+细胞百分比的作用明显优于CrvpPS、蔗糖-纳米硒及左旋咪唑(P<0.05).表明CrvpPS对免疫功能低下小鼠的胸腺指数、T细胞亚群和NK细胞百分比具有正向调节作用;其硒化产物CrvpPS-nano-Se对CD3+,CD3+CD4+细胞的调节作用优于单纯多糖和单纯纳米硒.

中国南海总状蕨藻的脂肪酸类化学成分和生物活性研究

目的分离鉴定中国南海总状蕨藻（Cauler pa rucennosu）中的脂肪酸类化学成分。供药理活性筛选。方法综合利用各种柱层析方法分离纯化化合物并利用光谱学方法解析化合物结构；采用M3-F法对所有化合物进行神经细胞损伤保护作用评价；采用酶标法对所有化合物进行蛋白酪氨酸磷酸酯酶抑制活性评价。结果从中分离得到12个脂肪酸及衍生物，分别鉴定为棕榈酸（1）、硬脂酸、（2）、十八皖二酯二乙酯、（3）、正二十三烷酸甘油酯（4）、（8E）-heplade0c-8-em7-one（5）、（8Z，11Z）-heptadeca-8，11-dienoicacid（6）、ethyl linolenoate（7）、（4Z，7Z．10Z，13Z）-ethyl hexadeca-4，7，10，13-telraenoate（8）、β-dimorphecolic acid（9）、α-adimorphecolic acid（10）、（9R，10E，12Z）-9-hydroxyoctadeca-10。12-dienoicacid（11）、（9R，10E）-9-hvdroxyoctadec-10-enoicacid（12）。结论化合物5为新天然产物；化合物3～12为首次从总状蕨藻中分离得到，其中6和7首次发现于蕨藻属海藻中，3，4和8～12均为首次从海洋生物中分离得到；化合物2和5在10μmol／L浓度时对SH—SY5Y神经母细胞瘤细胞损伤显示了不同强度的保护作用；化合物1，2，5，11和12对蛋白酪氨酸磷酸酯酶1β均显示了明显的抑制活性。

EFFECT OF LIGHT INTENSITY ON THE TOTAL LIPID AND FATTY ACID COMPOSITION OF SIX STRAINS OF MARINE DIATOMS

The effect of light intensity (1500 lx and 5000 lx) on the total lipid and fatty acid composition of six strains of marine diatoms Cylindrotheca fusiformis (B211), Phaeodactylum tricornutum (B114, B118 and B221) Nitzschia closterium (B222) and Chaetoceros gracilis (B13) was investigated. The total lipids of B13, B114, and B211 grown at 5000 lx were lower than those grown at 1500 lx. No evident changes were observed in B118, B221 and B222. Fatty acid composition changed considerably at different light intensity although no consistent correlation between the relative proportion of a single FA and light intensity. The major fatty acids of the 6 strains were 14:0, 16:0, 16:1(n-7) and 20:5(n-3). Cylindrotheca fusiformis had high percentage of 20:4n-6 (9.2-10.9%). The total polyunsaturated fatty acid in all 6 strains decreased with increasing light intensity. The percentage of the highly unsaturated fatty acid eicosapentaenoic acid (EPA) decreased with increasing light intensity in all strains except Chaetoceros gracilis.

EFFECTS OF HARVEST STAGE ON THE TOTAL LIPID AND FATTY ACID COMPOSITION OF FOUR CYLINDROTHECA STRAINS

Four strains of Cylindrotheca (B156, B164, B196, and B200) were harvested at three phases in the growth curve and their total lipid and fatty acid composition were determined. Total lipid reached highest values in the late stationary phase in B156 (24.0%), B164 (26.6%) and B200 (17.3%), in the exponential phase in B196 (15.5%). Saturated fatty acids increased with the development of the culture in B156 and B196, and reached their highest values in the late stationary phase in B156 (41.7%) and B196 (45.1%), in the early stationary phase in B164 (45.4%) and B200 (37.6%). Monounsaturated fatty acids increased with the development of the culture in B196 and B200, and reached their highest values in the late stationary phase in B196 (32.4%) and B200 (32.8%), in the early stationary phase in B164 (31.0%) and in the exponential phase in B156 (29.3%). Polyunsaturated fatty acids decreased in the later phases of the culture in B164, B196 and B200, and peaked in the exponential phase in B164 (29.5%), B196 (42.9%) and B200 (37.3%), and in the early stationary phase in B156 (32.0%).

其他

从63种海洋藻类和16种红树植物中筛选抗烟草花叶病毒(TMV)的种类,有16种藻类具有抗病毒活性,其中包括4种绿藻(总状蕨藻、棒叶蕨藻、匍匐松藻、大叶仙掌藻)、4种褐藻(两种网地藻、软叶马尾藻、圈扇藻)以及8种红藻(纵胞藻、仙菜、凝花菜、皮江蓠、钩沙菜、两种凹顶藻、多管藻)。

Application of Box-Behnken Design in Optimization of Allelopathic Effects of Potamogeton Pectinatus Against Microcystis Aeruginosa

Allelopathic effects of submerged macrophytes against algae are affected by many environmental factors which can only be measured one by one by traditional methods. Box-Behnken design of response surface methodology was used to optimize three environmental factors （temperature, light intensity and total dissolved solids） of allelopathic effects of Potarnogeton pectinatus against Microcystis aeruginosa at the same time. By solving the regression equation and analyzing the response surface contour plots, the optimal conditions of the relatively inhibitory rate of Microcystis aeruginosa were that the temperature was 23℃, the light intensity was 2 700 lx and the total dissolved solids were 4 415 mg/L. Under these conditions, the optimal value of relatively inhibitory rate of Microcystis aeruginosa was 81.9%. According to validation experiments, the results of analysis indicated that the experimental values fitted well with the predicted ones.

The Mechanism of Lipids Extraction from Wet Microalgae Scenedesmus sp. by Ionic Liquid Assisted Subcritical Water

In this paper, the total sugar concentration, protein concentration, lipid yield and morphology characteristics of the algae residue were determined to explain the mechanism of lipids extraction from wet microalgae Scenedesmus sp. by ionic liquid assisted subcritical water. The results showed similar variation for the sugar, protein and lipid. However, the total sugar was more similar to lipids yield, so the results showed that the reaction between ionic liquid and cellulose and hemicellulose in cell wall was the most important step which determined the lipids extration directly. And the total sugar variation may be representing the lipids yield. For later lipids extraction, we can determine the total sugar concentration to predict the extraction end product.

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on the Cholesterol-Lowering in Vitro and in Vivo

The survey＇s results showed a significant percentage of women and especially men having an above normal cholesterol. In order to help reduce excessive rate of blood cholesterol, we used a local strain of Spirulina （Tamanrasset, in south of Algeria） associated with probiotic bacteria （Bf adolescentis）. Experiments in vitro showed a significant degradation of total cholesterol by the combination of Spirulina and Bf adolescentis （74.5%） after 72 hours incubation at 37 ~C. A cholesterol is added to the standard diet mice in order to increase the total cholesterol for three lots. However, the rates of total cholesterol in mice receiving lower fermented milk with Bf adolescentis enriched by dry Spirulina. This shows that the decrease of cholesterol rate is closely related to the presence of Bf adolescentis and Spirulina. In vitro and in vivo results show, however, that it is possible to obtain a natural product （Spirulina） and a pseudo-strain lactic （Bf adolescentis） and to participate in the prevention of cardiovascular disease risk factor whose hand is cholesterol.

Antioxidant system responses in two co-occurring green-tide algae under stress conditions

Green tides have occurred every year from 2007 to 2014 in the Yellow Sea. Ulva prolifera （Mtiller） J. Agardh has been identified as the bloom-forming alga, co-occurring with U. intestinalis. We observed distinct strategies for both algal species during green tides. U.prolifera exhibited a high abundance initially and then decreased dramatically, while U. intestinalis persisted throughout. The antioxidant system responses of these two macroalgae were compared in the late phase of a green tide （in-situ） and after laboratory acclimation. Lipid peroxidation and antioxidant system responses differed significantly between the two. Malondialdehyde and hydrogen peroxide contents increased significantly in-situ in U. prolifera, but not in U. intestinalis. In U. prolifera, we observed a significant decrease in total antioxidant ability （T-AOC）, antioxidant enzymes （SOD and Apx）, and non-enzyme antioxidants （GSH and AsA） in-situ. U. intestinalis showed the same pattern of T-AOC and SOD, but its Gpx, Apx, and GSH responses did not differ significantly. The results suggest that U. prolifera was more susceptible than U. intestinalis to the harsh environmental changes during the late phase of a Yellow Sea green tide. The boom and bust strategy exhibited by U. prolifera and the persistence of U. intestinalis can be explained by differences in enzyme activity and antioxidant systems.

Purification and Antioxidant Activities of Phycocyanin from Spirulina Platensis

This study aimed to purify and determine antioxidant activities of different fractions obtained during the purification process of phycocyanin from Spirulina platensis. The dried powder of Spirulina platensis, after ground with sands, was extracted with 50 mM sodium phosphate buffer at pH 6.8 before centrifuged to precipitate unwanted proteins. Then the supernatant was separated by celit column to obtain semi-pure phycocyanin and further purified by treated with ammonium sulfate. The purity of phycocyanin was monitored by measuring the absorbance spectrum from 200 to 700 nm. Its purity ratio A620A280 was determined. The antioxidant activities of the obtained phycocyanin were determined by 2,2＇-azino-bis（3-ethylbenzthiazoline-6-sulphonic acid） （ABTS） assay and lipid peroxidation （linoleie acid） assay. The purity ratio of phycocyanin in the Spirulina crude extract was 0.36 and increased to 2.68 after purification. The fraction with the highest purity ratio of phycocyanin demonstrated the hightest antioxidant activities. For ABTS assay, it presented the Vitamin C Equivalent Antioxidant Capacity （VCEAC） value of 0.0405 ±0.0002 mg of ascorbic acid/mg of sample and the Trolox Equivalent Antioxidant Capacity （TEAC） value of 0.0485 ±0.0002 mg oftrolox/mg of sample respectively. The result from lipid peroxidation assay exhibited IC50 value of 5.9336 ±0.2565 mg/mL. The purification of Spirulinaplatensis crude extract obtained from this study increased the purity ratio of phycocyanin and its antioxidant activities. This will be further investigated for the development into anti-aging cosmetic products.

勘误

对《动物学杂志》2022年57卷5期第775至781页发表的论文《海南省首次发现光合软体动物绒毛海天牛》进行勘误。该文中5处总状蕨藻的学名“Caulerpa racemose”应更改为“Caulerpa racemosa”,分别位于:(1)775页中文摘要部分第5行;(2)776页第1行;(3)778页图1a英文图注;(4)779页图3英文图注第2行;(5)779页正文左栏第6行。

Antioxidant activity of alcohol aqueous extracts of Crypthecodinium cohnii and Schizochytrium sp.

Crypthecodinium cohnii(dinoflagellate) and Schizochytrium sp.(thraustochytrid) are the main sources for docosahexaenoic acid(DHA). The present study aimed to evaluate the antioxidant activity of petroleum ether, ethyl acetate, n-butanol, and water fractions of alcohol aqueous extracts of these two microalgae and to provide a theoretical basis for comprehensive utilization. The antioxidant activity was determined by total antioxidant capacity(TAC) determination, 2,2-diphenyl-1-picrylhydrazyl(DPPH) radical scavenging assay, ferrous ion-chelating ability(FICA) assay, and reducing power(RP) assay. The total phenolic content(TPC) and total flavonoid content(TFC) were also measured by the Folin-Ciocalteu and spectrophotometry methods, respectively. The results indicated that the extracts from these two microalgae possessed good antioxidant capacity. Analysis showed that most antioxidant performance indicators(TAC, DPPH, and RP) were positively correlated with the TPC of the extracts, suggesting that the phenolics might be the major components in C. cohnii and Schizochytrium sp., contributing to their antioxidative function. Therefore, the polar fractions of C. cohnii and Schizochytrium sp. could be further examined and considered for application in health products or cosmetics.