C-phycocyanin shows neuroprotective effect against rotenone-induced Parkinson’s disease in mice

Objective:To investigate the neuroprotective effect of C-phycocyanin in a mouse model of rotenone-induced Parkinson’s disease.Methods:C-phycocyanin(50 mg/kg,i.p.,daily)was administered to rotenone(30 mg/kg,p.o.,daily)treated mice for 28 days.Behavioral studies(Y-maze,rotarod,round beam walk,and wire-hang tests)were carried out to assess neurobehavioral deficits.Glutathione and malondialdehyde were determined in both serum and striatal tissue.Molecular proteins(AKT,AMPK,NF-κB,BDNF,and alpha-synuclein)in the striatum were estimated using ELISA.Histopathological analyses(hematoxylin and eosin stainning as well as Nissl staining)were carried out to assess structural abnormalities in the striatum.Results:C-phycocyanin significantly increased BDNF levels and decreased alpha-synuclein levels.It also slightly upregulated AMPK and AKT levels without significant difference compared with the rotenone group.Additionally,rotenone-induced elevated oxidative stress and structural abnormalities in the striatum were markedly mitigated by C-phycocyanin.Conclusions:C-phycocyanin might have potential neuroprotective effects against Parkinson’s disease.Further studies are warranted to verify its efficacy and to understand the molecular mechanisms behind the neuroprotective effects of C-phycocyanin in Parkinson’s disease.

The Stability of C-phycocyanin Doped Silica Biomaterials in UV Irradiation

The synthesized C-phycocyanins （C-PCs） doped silica biomaterials were characterized by the SEM and BET surface area analysis measurement. The morphology of C-PCs doped silica biomaterials indicates that the surface of the silica cluster is formed by a great number of silica particles with an average size of between 30 and 40 nm. Silica itself is a porous structure with the average pore diameter of 2.95 nm. Pores with their diameter less than 5 nm account for 84.07%. In addition, the C-PCs can be utilized as a fluorescent protein probe to monitor influence of the protein encapsulation and to study matrix and protein interaction and stability of protein in silica matrix. Application of protein encapsulation silica materials requires biomolecules to keep bioactivity and stability on potentially unfavorable industrial conditions. The C-PCs in solution or in silicate matrix irradiated by ultraviolet ray can result in photobleaching, whereas the protein in the silica is less affected. The measured photodamage rate constant of C-PCs in buffer solution is 25 times faster than that of C-PCs in silica matrix. However, the lifetime of C-PCs in silica matrix or phosphate buffer is unaffected. These studies suggest that entrapment of C-PCs into silica matrixes not only can maintain their biological activity but also noticeably improve their photostability.

Protective Role of C-Phycocyanin Against Secondary Changes During Sodium Selenite Mediated Cataractogenesis

Age related cataract is the leading cause of blindness associated with accumulation of oxidative stress in the eye lens.The present investigation reveals the rational of the beneficial effects of the natural compound C-phycocyanin(CPC)is beneficial when administered to rat pups to protect against the secondary effects of sodium selenite induced cataractogenesis.A single subcutaneous dose of sodium selenite(19 lmol/kg body weight)on the 10th day of postpartum is adequate to induce cataract in rat pups.Serum biochemical parameters,such as the level of electrolytes,mean activities of anti-oxidant enzymes i.e.superoxide dismutase,catalase and reduced glutathione were observed to be significantly altered during selenite induced cataractogenic process.Histopathological examination revealed signs of degradation of normal cell architecture in the liver,kidney and eye lens.Interestingly,the deleterious effects of sodium selenite toxicity were restored with the simultaneous treatment with C-PC.The results suggest that an administration of 200 mg/kg body weight of C-PC has the ability to prevent/alter the secondary changes reflected in the serum biochemical and histological modifications in rats exposed to sodium selenite.These results complement the beneficial role of C-PC of cyanobacterial origin as a efficacious anti-cataractogenic agent against sodium selenite toxicity.

Protective effects of C-phycocyanin on alcohol-induced acute liver injury in mice

Excessive alcohol consumption leads to liver disease. Extensive evidence suggests that C-phycocyanin(C-PC), a chromophore phycocyanobilin derived from Spirulina platensis, exerts protective eff ects against chemical-induced organ damage. In this study, we investigated whether C-PC could protect against ethanol-induced acute liver injury. Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), triglyceride(TG), total cholesterol(CHOL), low-density lipoprotein(LDL), liver homogenate malondialdehyde(MDA), superoxide dismutase(SOD) content were measured, and pathological examination of liver sections were examined. C-PC showed obvious inhibitory eff ects on serum ALT, AST, TG, CHOL, LDL and MDA, and SOD content significantly increased in the liver. The structure of hepatic lobules was clear, liver sinus returned to normal, and liver cell cords were arranged in neat rows. Cloudiness, swelling, inflammatory cell infiltration and spotty necrosis of liver cells were significantly reduced. Therefore, C-PC can significantly protect against ethanol-induced acute liver injury.

Effect of Hg(II) and Pb(II) Ions on C-Phycocyanin (<i>Spirulina</i><i>platensis</i>)

Influence of Hg(II) and Pb(II) ions on C-Phycocyanin (C-PC) from cyanobacteria Spirulina platensis was investigated using Fluorescence spectroscopy. Fluorescence measurements demonstrate quenching of C-PC emission by Hg(II) and Pb(II), and blue shifts in the fluorescence spectra. The effect of DNA on the fluorescence of Hg(II)-and Pb(II)-C-PC (from Spirulina platensis) complexes was also studied. It was shown that the fluorescence intensity of Hg-C-PC after addition of DNA gave rise to the fluorescence buildup. At the same time, addition of DNA to the Pb(II)-C-PC complexes showed no such effect. In the case of Hg(II)-C-PC, fluorescence intensity significantly decreases in time, while for Pb(II)-C-PC, decrease of the fluorescence intensity is not significant, but blue shift of the peak takes place.

Isolation and Characterization of C-phycocyanin from Spirulina Platensis

The isolation of biliproteins from the Spirulina platensis cultured in southern China was accomplished with gel filtration on Sephacryl S 200 and chromatography on hydroxylapatite. The spectrophotometry, isoelectric point, and amino acid composition of C phycocyanin were determined, respectively.

Construction of energy transfer model of C-phycocyanin and allophycocyanin from Spirulina platensis

PHYCOBILIPROTEINS. which include phycoerythrin, phycocyanin, phycoerythroeyanin and allophycocyanin, are intensively coloured, highly fluorescent, water soluble chromoproteins. They are composed of at least two dissimilar polypeptides, α and β subunits, in a molar ratio of 1:1. The subunits possess linear tetrapyrrole prosthetic groups covalently linked to the primary structure via cysteine thioether linkages. In blue-green and red algae, phycobiliproteins occur in a supramolecular structure, the phycobilisome, located on the surface of thylakoid to

Kinetics of energy transfer processes in C-phycocyanin complexes

The antenna system of algae for photosynthesis is a functional entity composed of various phycobiliproteins and the linker polypeptides. Up to now, high-resolution crystal structure data have been available only for the isolated phycohiliproteins. To have an understanding of the functional connection between different phycobiliproteins, it is necessary to study the complexes composed of different phycobiliproteins. The energy transfer processes in C-phycocyanin complexes were studied through computer simulation because it is dificult to be studied by conventional experimental methods. The main pathways of energy flow and the dynamic property of the energy transfer were obtained. A fast transfer process between two neighboring disks was observed through analyzing the distribution curves of excitation energy over time. According to the definition of the time constants for energy transfer in time-resolved spectrum techniques, for a complex with three C-phycocyanin hexamer disks, a fluorescence-rising component with several picoseconds was obtained in disk 2 and 20 ps in disk 3 when the chromophores in disk 1 were excited by an ideal Δ pulsed laser beam. The results showed clearly that the energy transfer between the disks in an antenna rod was very fast and that energy transfer was fast and efficient in an intact antenna system of algae.

Energy transfer from C-phycocyanin to phthalocyanine metal complex in reverse micelles

A new mimic system of photosymhetic apparatus was constructed from C-phycocyanin and phthalocyanine zinc. C-FC was solubilized in the reverse micelles of non ionic surfactant Tween-80, cosurfactant pentanol, and solvent cyclohexane, in which the overall concentration of surfactant was 20% (w/v) and the mass ratio of Tween-80 to pentanol was 4:1. When the molar ratio of water to Tween-80 (R w)?S9.0, the characteristic properties of C-PC were maintained. When it was excited, the energy transfer from C-PC to phthalocyanine zinc took place. The energy transfer efficiency was only related with the concentration of phthalocyanine, but not that of C-PC. Furthermore, the energy transfer was roughly in keeping with Perrin formulation, which indicated that the energy transfer took place approximately through dipole-dipole interaction in rigid system. The radii of the quenching sphere were calculated from the experimental results. For example, when the concentration of phthalocyanine zinc was 2.10×10-4 mol/L, the radius of the quenching sphere of the system was 10.9 nm.

SPECTROSCOPIC CHARACTERISTICS AND ENERGY TRANSFER PROCESSES IN C-PHYCOCYANIN FROM CYANOBACTERIUM Westiellopsis prolifica

The spectroscopic characteristics and pathways of energy transfer in the C--phycocyaninmonomer (αβ) and trimer(αβ)_3 isolated from cyanobacterium Westiellopsis prolifica havebeen investigated by absorption, excitation, fluorescence, fluorescence excitation polarizationand fluorescence polarization spectra and the deconvolution of those spectra. The phenome-non of an energy--back--transfer within a C--phycocyanin trimer has been found, and a tentativemodel of the bidirectional energy transfer is proposed by us, which describes well our exper-imental results.

C-藻蓝蛋白纳米微球的制备及体外抗脂多糖联合海水诱导急性肺损伤的作用机制研究

目的制备C-藻蓝蛋白纳米微球(CPC-NPs)并评价其对脂多糖(LPS)联合海水诱导急性肺损伤的体外作用机制。方法采用离子交联法,以CPC为药物、羧甲基壳聚糖(CMCS)为载体、CaCl2为交联剂制备CPC-NPs,对CPC-NPs进行基础表征。小鼠肺泡Ⅱ型上皮细胞MLE-12和巨噬细胞RAW264.7均分为7组:正常组(Con组),模型组(Mod组),空白NPs组,CPC-NPs 30、60、120、240μg/mL组。除Con组外其余各组均采用10μg/mL LPS和25%海水联合处理6 h,造模后各给药组加相应剂量药物处理24 h。检测MLE-12细胞中丙二醛(MDA)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)水平,B细胞淋巴瘤2(Bcl-2)、Bcl-2关联X蛋白(Bax)、胱天蛋白酶-3(caspase-3)蛋白和mRNA,以及CAT、谷胱甘肽S-转移酶(GST)mRNA表达水平;检测RAW264.7细胞中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-6水平,NOD样受体热蛋白结构域相关蛋白3(NLRP3)、剪切的caspase 1(cleaved-caspase-1)蛋白及IL-1β、TNF-α、IL-6、诱导型一氧化氮合酶(iNOS)mRNA表达水平。结果制备的CPC-NPs粒径为(675.69±64.58)nm,Zeta电位为(-20.11±0.98)mV,多分散系数为0.455±0.010(n=3);包封率为35.60%,载药量为16.13%;形态为规则的球形;其中药物可持续释放30h以上。与Mod组比较,CPC-NPs各浓度组MLE-12细胞中T-AOC、SOD、CAT(除CPC-NPs 30μg/mL组外)、GSH-Px水平和CAT、GST mRNA表达水平以及Bcl-2/Bax蛋白比值和mRNA比值均显著升高,MDA水平和caspase-3蛋白及mRNA表达水平均显著降低(P<0.01或P<0.05)。与Mod组比较,CPC-NPs各浓度组RAW264.7细胞中IL-1β、TNF-α、IL-6水平和NLRP3、cleaved-caspase-1蛋白表达水平以及IL-1β、TNF-α、IL-6、iNOS mRNA表达水平均显著降低(P<0.01或P<0.05)。结论成功制备了具有肺靶向性和缓释性的CPC-NPs。CPC-NPs可以通过抗氧化应激及抑制细胞凋亡和炎症反应来缓解LPS联合海水诱导的急性肺损伤。

活性炭对极大节旋藻来源藻蓝蛋白和蛋白质的吸附性能和机制研究

通过等温吸附、吸附动力学和吸附热力学试验,揭示活性炭对C-PC和总蛋白质的吸附特性和作用机制。结果显示:同样条件下,活性炭去除杂蛋白质的效率更高,经过活性炭纯化后,C-PC占总蛋白质的比例从31.2%增加到51.5%;活性炭对C-PC和总蛋白质的最大吸附量分别为10.5和88.5 mg/g;活性炭对C-PC的吸附为单分子层化学吸附,符合Langmuir等温吸附模型,而对总蛋白质的吸附为多层级的吸附,符合Freundlich等温线模型。

藻蓝蛋白对小鼠皮肤损伤的治疗效果

为探讨藻蓝蛋白(CPC)对小鼠皮肤损伤的治疗效果,将ICR小鼠分为正常对照组(Con)、模型组(Mod)、空白敷料组(BD)、藻蓝蛋白敷料组(CPC TDD)和藻蓝蛋白敷料+藻蓝蛋白腹腔注射组(CPC+ip.)5个处理组进行小鼠烫伤试验。结果表明,CPC TDD组和CPC+ip.组伤口愈合时间最短,可以降低血清中丙二醛(MDA)含量,提高超氧化物歧化酶(SOD)含量,降低血清中炎症因子TNF-α和IL-6的水平。藻蓝蛋白通过增强机体抗氧化能力,降低炎症因子水平,从而对皮肤损伤的愈合起到促进作用。

钝顶螺旋藻藻蓝蛋白的富集分离及其稳定性研究

实验研究了盐析沉淀法和等电点沉淀法富集分离钝顶螺旋藻藻蓝蛋白,考察了螺旋藻细胞破碎液的藻液比、盐离子浓度、pH和环境温度对藻蓝蛋白收率和分离因数的影响,并对藻蓝蛋白的热稳定性及酸碱稳定性进行了研究。结果表明,藻液比为0.5g/100ml,50%(W/V)(NH4)2SO4进行盐析沉淀,藻蓝蛋白收率可达60.0mg/g藻粉,分离因数为1.38。螺旋藻藻蓝蛋白在20℃和30℃时保持稳定,在40℃以上稳定性下降。在pH为中性时,藻蓝蛋白稳定性良好,超出此范围螺旋藻藻蓝蛋白稳定性下降。

螺旋藻中藻胆蛋白和多糖的分离纯化及产品特性

提出一种从螺旋藻中提取纯化藻蓝蛋白和糖蛋白的新型分离工艺。等电聚焦分析结果表明所得到的藻蓝蛋白为电泳纯，等电点为４．３，测定了藻蓝蛋白的氨基酸组成及其红外吸收光谱。ＳＤＳ－ＰＡＧＥ分析结果表明所得到的糖蛋白组成均一，其分子量为１３０ｋｄ，测定了螺旋藻糖蛋白的氨基酸组成，红外光谱分析表明螺旋藻糖蛋白含有α－型糖苷连接键。

钝顶螺旋藻藻蓝蛋白诱导HeLa细胞凋亡的分子机制研究

目的探讨钝顶螺旋藻（Spirulina platensis）藻蓝蛋白（C-phycocyanin,CPC）对体外人宫颈癌细胞（HeLa细胞）凋亡的影响及分子机制。方法首先通过MTT法检测藻蓝蛋白对HeLa细胞增殖的影响,然后通过电镜观察藻蓝蛋白处理后的细胞的凋亡形态,琼脂糖凝胶电泳观察凋亡细胞的DNA结构变化,流式细胞仪显示不同浓度藻蓝蛋白对HeLa细胞的细胞周期变化的影响,免疫组化法检测藻蓝蛋白对细胞表面凋亡相关基因表达的影响,检测凋亡相关的caspase的活性及细胞色素C的释放,以揭示藻蓝蛋白诱导HeLa细胞凋亡的作用机制。结果与未用藻蓝蛋白作用的对照组相比,藻蓝蛋白组的HeLa细胞存活率明显降低,并存在浓度剂量效应。藻蓝蛋白处理后的细胞呈现典型的凋亡形态,如细胞皱缩、细胞膜微绒毛消失、起泡,染色体密集,形成凋亡小体。凋亡细胞的DNA出现断裂,呈梯状的电泳带（DNAladder）,分子量为180~200 bp及其倍体。并且发现藻蓝蛋白处理后的处于亚二倍体峰HeLa细胞数蛋白浓度随藻蓝蛋白浓度的增高而明显增多。另外藻蓝蛋白能促进HeLa细胞内促凋亡基因（Fas、ICAM-1）的表达并抑制抗凋亡基因（Bcl-2）的表达,从而促进细胞内凋亡信号的传导。藻蓝蛋白可以激活caspase酶并可促进凋亡蛋白——细胞色素C从线粒体到细胞质释放。结论钝顶螺旋藻藻蓝蛋白具有诱导体外HeLa细胞凋亡的作用,通过促进凋亡信号在HeLa细胞内的传导从而达到抗肿瘤的效果。

藻蓝蛋白对小鼠粒单系祖细胞生成的影响

藻蓝蛋白(C—phycocyanin,C—PC)是从钝顶螺旋藻(Spirulina platensis)中分离、纯化所得。采用造血祖细胞体外培养技术,研究了C—PC对小鼠粒单系祖细胞生成的影响。实验结果表明,C—PC能够提高粒单系祖细胞(CFU—GM)的生成,亦能明显提高正常小鼠血清的集落刺激活性。C—PC体内或体外刺激制备的小鼠脾细胞条件培养液能明显增加CFU—GM的集落数。在有集落刺激因子(GM—CSF)存在时,体外加入C—PC培养粒单系祖细胞,C—PC能明显提高CFU—GM的集落生成率。

双水相萃取法富集分离螺旋藻藻蓝蛋白的研究

以PEG/硫酸钠双水相体系,经一次萃取从钝顶螺旋藻(Spirulina platensis)细胞破碎液中富集分离藻蓝蛋白。结果表明,萃取最适宜的条件为12%PEG4000,15%Na2SO4,1%KCl,藻蓝蛋白收率为91.2%,分配系数达到8.01,分离因数达到6.33。对于螺旋藻藻蓝蛋白的富集分离,双水相萃取法与传统的盐析沉淀法相比,具有节省操作时间、简化操作过程、降低能耗和成本以及易于工艺放大等优点。

从一种富含藻胆蛋白的螺旋藻中大量提取和纯化藻蓝蛋白的研究

采用新鲜藻丝为原料和分段梯度盐析分离纯化钝顶螺旋藻 Sp(NS) - 90 0 2 0的藻蓝蛋白 (PC) ,经羟基磷灰石一次层析 ,能使提取的 PC的纯度大于普遍认可的标准。由于该工艺流程较为简单 ,适合藻蓝蛋白的大量生产。藻胆蛋白经 Sephadex凝胶过滤后的可达电泳纯度标准。经 SDS- PAGE测得 PC,APC的分子量分别约为 38,33k

藻蓝蛋白介导光动力疗法在乳腺癌治疗中的机制研究

目的探讨藻蓝蛋白介导的光动力学疗法在乳腺癌治疗中的机制。方法将MCF-7细胞接种于小鼠肋缘皮下脾区构建乳腺癌小鼠模型。小鼠分成4组:对照组、He-Ne激光照射组、藻蓝蛋白处理组、光动力学治疗组(PDT组)。10d后检测瘤块重量,NK细胞活性和T细胞增殖活性。取瘤块制成石蜡包埋切片,采用原位核酸杂交技术、免疫组织化学技术检测组织细胞内凋亡相关蛋白的表达。体外培养的MCF-7细胞也相应分成4组,通过MTT法、电镜和免疫荧光技术检测细胞增殖活性、细胞形态、细胞色素C表达量的变化。结果与对照组相比,激光照射组各检测指标均无明显差异,而藻蓝蛋白处理组中NK细胞和T细胞的增殖活性有所增强,肿瘤组织细胞内抗凋亡蛋白(Fas)表达量明显增多,而瘤块的重量、肿瘤形成率和抗凋亡蛋白(p53、NF-κB、CD44)明显减少,如果藻蓝蛋白结合激光治疗发现各检测指标与对照组比较差异更为明显。体外试验证实藻蓝蛋白能抑制MCF-7细胞的增殖,促进细胞色素C的释放,电镜下细胞呈现典型的凋亡形态,用光动力学方法处理后效果更为明显。结论藻蓝蛋白可以作为一种光敏剂,其介导的光动力学疗法通过增强机体的免疫力同时启动乳腺癌细胞内的凋亡信号转导通路诱导细胞凋亡,从而达到杀死肿瘤的目的。