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@# Objective: To investigate the effect of C-phycocyanin (C-PC) on the epithelial-mesenchymal transition (EMT) of cervical cancer Caski cells induced by transforming growth factor beta1 (TGF-β1). Methods: According to different treatment methods, Caski cells were divided into three groups: 10 ng/ml TGF-β1 treatment group, 10 ng/ml TGF-β1+300 μg/ml C-PC co-treatment group and control group (untreated). After 24 h of treatment, the morphological changes of Caski cells were observed, and the effects of TGF-β1 and C-PC on the migration and invasion of Caski cells were detected by Scratch test and Transwell test, respectively. Western blotting was used to detect the effect of C-PC on the expression of epithelial phenotypic marker protein E-cadherin and stromal phenotypic marker protein N-cadherin in TGF-β1-induced Caski cells, and qPCR was used to detect the mRNA expressions of EMT related factors Snail, Zeb1 and Twist. Results: Caski cells in the TGF-β1 treatment group lost the characteristics of the original epithelial phenotype, while the cells in the TGF-β1+C-PC co-treatment group maintained the characteristics of normal epithelial phenotype; the migration rate ([60.0±1.4]% vs [33.5±2.2]%, [40.0±2.8]%, both P<0.05) and the number of invasive transmembrane cells ([108.2±6.2] vs [25.2±3.1], [39.8±5.4], both P<0.01]) of Caski cells in the TGF- β1 treatment group were significantly higher than those in the co-treatment group and the control group. Compared with the control group, the expression of E-cadherin in Caski cells treated with TGF-β1 decreased significantly (P<0.05), while the mRNA expressions of Twist, Snail and Zeb1 increased significantly (all P<0.05); However, co-treatment with C-PC reversed above changes (P<0.05 or P<0.01), and significantly decreased the protein expression level of N-cadherin (P< 0.05). Conclusion: C-PC treatment can inhibit the invasion and metastasis ability of Caski cells induced by TGF-β1 and further affects the EMT process. The mechanism may be related to the decrease of mRNAexpressions of Twist, Snail and Zeb1 by C-PC treatment. ·
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Las ficobiliproteínas son proteínas solubles en agua, que funcionan como pigmentos fotosintéticos accesorios en diferentes organismos tales como las cianobacterias, las algas rojas y las criptomonadas. En el alga verdeazul Spirulina platensis, una de las ficobiliproteínas más abundantes es la C-ficocianina, la cual tiene unido tres cromóforos ficocianobilina mediante un enlace tioéter a cisteínas específicas. La ficocianobilina es un tetrapirrol lineal asociado a la captación de energía solar en estos organismos. La C-ficocianina ha sido empleada en diferentes investigaciones biomédicas como biomarcador, por sus propiedades fluorescentes, y como posible agente terapéutico para el tratamiento de enfermedades asociadas al estrés oxidativo, por sus propiedades antioxidantes, inmunomoduladoras y antinflamatorias. Se ha demostrado que esta proteína aumenta la liberación de interferón gamma en células mononucleares de sangre periférica y modula la producción de citocinas inflamatorias como el factor de necrosis tumoral alfa, entre otras. Además, se ha encontrado que la C-ficocianina tiene efecto inmunomodulador de citocinas que potencian la activación de las células del sistema inmune, como la IL-6 y la IL-1ß, así como la regulación de aproximadamente 190 genes implicados en la inmunidad(AU)
Phycobiliproteins are water-soluble proteins that function as accessory photosynthetic pigments in different organisms such as cyanobacteria, red algae and cryptomonads. In the blue-green algae Spirulina platensis one of the most abundant phycobiliproteins is the C-phycocyanin, which has three phycocyanobilin chromophores linked through a thioether bond to specific cysteine. The phycocyanobilin is a linear tetrapyrrole associated with solar energy absorption in these organisms. The C-phycocyanin has been used in several biomedical researches as a biomarker, for their fluorescence properties, and as a possible therapeutic agent for the treatment of diseases associated with oxidative stress for its antioxidant, anti-inflammatory and immunomodulatory properties. It has been shown that this protein increases the release of interferon gamma in peripheral blood mononuclear cells, and modulates the production of inflammatory cytokines such as tumor necrosis factor among others. Furthermore it has been found that the C-phycocyanin has immunomodulatory effect on cytokines that enhance the activation of immune cells, such as IL-6 and IL-1ß, and the regulation of about 190 genes involved in immunity(AU)
Subject(s)
Phycobiliproteins/therapeutic use , Immunologic Factors/therapeutic use , Phycocyanin/therapeutic useABSTRACT
C-phycocyanin from Spirulina platensis was purified in aqueous two-phase systems (ATPS) of polyethylene glycol (PEG)/potassium phosphate, varying the molar mass of the PEG. Results using a full factorial design showed that an increase in the concentration of salt and decrease in the concentration of PEG caused an increment in the purification factor for all the ATPS studied. Optimization of the conditions of the purification was studied using a central composite rotatable design for each molar mass of PEG. The ATPS composed of 7% (w/w) PEG 1500 or 4% (w/w) PEG 8000 (g/gmol) and 23 or 22.5% (w/w) of phosphate resulted a purification factor of 1.6-fold for C-phycocyanin, with total and 57% recovery, respectively. Process conditions were optimized for the purification factor for the system with PEG 1500. The ATPS with 4% (w/w) PEG 4000 or 4% (w/w) PEG 6000 and 21% (w/w) phosphate resulted purification factors of 2.1 and 2.2-fold, recovering 100% and 73.5%, respectively of C-phycocyanin in the top phase.
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Objective To observe the protective effect and molecular mechanism of C-phycocyanin (CPC) on acute lung injury (ALI) in septic rats. Method 75 SD rats were randomly divided into control group, model group and CPC group. Cecal ligation and puncture was used to establish a septic acute lung injury rats (model group). For the CPC groups, septic acute lung injury rats were administrated by 20, 40 and 60 mg/kg CPC by intraperitoneal injection. 72 h after the operation, serum and lung tissue were obtained, the wet to dry weight ratio, the content of TNF-α、IL-6 and IL-10 in bronchoalveolar lavage fluid, the activity of myeloperoxidase (MPO) was analyzed. Expression of heme oxygenase (HO)-1,activation of nuclear factor erythroid 2-related factor 2 (Nrf 2) and nuclear factor-kappa B (NF-B) were detected by Western blot. Superoxide and Nitrite/Nitrate Level production in Lungs and bronchoalveolar lavage fluid were measured by chemiluminescence and reduction method, respectively. Results Treatment with CPC significantly inhibited septic-induced inflammatory responses including elevation of superoxide formation, myeloperoxidase activity, leucocytes and protein infiltration in lung tissues, and production of proinflammatory cytokine, and nitrite/nitrate in bronchoalveolar lavage fluid (P<0.05). In addition, CPC could activate Nrf 2 and induce HO-1 expression, and inhibit NF-B activation in ALI rats. However, blocking HO-1 activity by tin protoporphyrin IX (SnPP), an HO-1 inhibitor, markedly abolished these beneficial effects of CPC in septic-induced ALI. Conclusion The protection mechanism of CPC may be through HO-1 induction and suppressing of NF-kB-mediated inflammatory responses.
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Objective To observe the role of heme oxygenase (HO)-1 on the protective effect of C-phycocyanin (CPC) on doxorubicin (DOX)-induced myocardial cells injury by Nvf2/HO-1 pathway. Methods 60 SD rats were randomly divided into control group, DOX group, CPC group and tin protoporphyrin IX (SnPP, an inhibitor of HO-1) group. The control group was injected with normal saline injection,while the DOX group was administrated with doxorubicin by intraperitoneal injection in a cumulative dose of 15 mg/kg for two weeks. For the CPC rats, 20, 40 and 60 mg/kg of CPC was administrated. The level of creatine kinase (CK) and lactate dehydrogenase (LDH) were detected, and the activity of HO-1 and caspase-3 were also examined. Expression of HO-1 and activation of Nrf 2 were detected by Western blot. Results Compared with control group, serum levels of CK, LDH and Caspase-3 activity in DOX group were significantly increased(P<0.05), but HO-1 in cardiac muscle was only increased slightly. upregulation. Treatment with CPC could significantly ameliorated the CK, LDH and Caspase-3 activity, and markedly induce HO-1 expression and its activity. The reduction of CK, LDH and Caspase-3 activity by CPC could be reversed by treatment of the HO-1 inhibitor, SnPP. Furthermore, CPC sould also induce Nrf 2 activation. Conclusion The protective effect of CPC on doxorubicin-induced myocardial cells in jury via Nrf 2 induced HO-1 HO-1 expression.
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C-Phycocyanin (C-PC) is a phycobiliprotein found in blue green algae, such as Spirulina platensis, is often used as a dietary nutritional supplement and exhibit a variety of pharmacological properties. In this regard, extraction, partial purification and antioxidant, anticoagulation and prevention of DNA damage activity of C-PC was investigated. In the present study, a simple and efficient method to extract C-PC from Spirulina platensis dry powder is reported. The extractions were carried out using two different methods: cold maceration and sonication method. The extraction using cold maceration method proved to be the most efficient method. Obtained crud CPC was purified by ammonium sulphate precipitation, dialysis and gel filtration and presented a final extraction yield of 3.27±0.09 mg/ml with a purity ratio of 2.317±0.08. When it was evaluated as an antioxidant in vitro, it was able to scavenge nitric oxide. C-PC showed significant anticoagulation and prevention of DNA damage activity.
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Isolation of three different active peptides from C-phycocyanin (C-pc) β chain of S. fussiformis and their biological properties are reported. Phycocyanin peptide β fraction 2 (cyanopeptide β 2) facilitated both antioxidant and plasmid DNA strand scission prevention activity due to higher cysteine moieties in the isolated peptide. The peptide significantly scavenged the free radicals like 1-1,-diphenyl-2-picryl hydrazyl and ferric reducing ability of plasma, increased the absorbance values in reducing power and also showed the higher trolox equivalent antioxidant capacity values in total reactive antioxidant potentials assay. Cyanopeptide β 2 also inhibited reactive oxygen species induced DNA pBR322 damage in dose dependent manner along with free radical scavenging properties suggesting the role in the DNA integrity which is also evident by DNA binding activity of peptide. In addition, the generation of reactive oxygen species (ROS) was dose dependent (10 and 20 ng/ml) and significantly quenched by cyanopeptide β2 in human fibroblast cell line TIG 3-20. In vitro cell scratch injury assay demonstrated the capacity of cyanopeptide β2 in cell migration in to wounded area suggesting fibroblast proliferation and migration. The results suggest that cyanopeptide β2 can be a free radical scavenger and effective peptide for future biomedical applications like wound healing, atherosclerosis, cell redox potential and hypoxia.
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Aim To investigate the influence and molecular mechanism of C-phycocyanin(CPC) from Spirulina platensis on apoptosis of HeLa cells in vitro.Methods Firstly,the effect of purified CPC on proliferation of HeLa cells in vitro was determined by MTT assay,and then electron microscope was exploited to observe the characteristic apoptotic features of cells treated with CPC.Subsequently,genomic DNA changes of HeLa cells were observed by agarose electrophoresis.Flow cytometric analysis revealed the influence of different concentrations of CPC on cell cycle of HeLa cells.The expressions of apoptosis related genes of CPC treated HeLa cells were determined by immunohistochemistry analysis.In addition,the activities of caspases and the release of cytochrome c from mitochondria into the cytosol were detected.Results Compared with control cells untreated with CPC,a significant decreased in the numbers of HeLa cells in survival treated with CPC and concentration dose effects existed.CPC could induce characteristic apoptotic features including cell shrinkage,membrane blebbing,microvilli loss,chromatin margination and condensation into dense granules or blocks.DNA of HeLa cells treated with CPC showed fragmentation pattern(DNA ladder of oligomers of 180~200 bp) typical for apoptotic cells.HeLa cells treated with different concentrations of CPC demonstrated an increasing percentage of cells in sub-G0/G1 phase.In addition,CPC could promote the expression of pro-apoptotic gene(Fas and ICAM-1);meanwhile,held back the anti-apoptotic gene(Bcl-2) expression,and then facilitated the transduction of tumoral apoptosis signals that resulted in the apoptosis of HeLa cells in vitro.In CPC treated HeLa cells,CPC treatment of HeLa cells also resulted in activation of caspases and release of cytochrome C from mitochondria into the cytosol.Conclusion C-phycocyanin from Spirulina platensis can induce the apoptosis of HeLa cells in vitro.By virtue of the promotion of the apoptosis signals transduction in HeLa,CPC realizes its antitumor activities.
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To investigate the effects of C-phycocyanin(C-PC) from Spirulina platensis on erythropoiesis in mice.The colony forming uniterythroid(CFU-E) and burst forming unit-erythroid(BFU-E) were determined using micro-methycellulose culture method in vitro.The erythropoietin(EPO)-like activity of C-PC was examined using the technique of CFU-E culture of fetal liver cells in mice in vitro.Anemic mice models were established by ~(60)Co ?-ray irradiation(5Gy) and intra-peritoneal injected with benzohyarazine hydrochloride.After normal mice were intra-peritoneal injected with C-PC (50 mg?kg~(-1)) for 5d,C-PC provided the only increase in the numbers of CFU-E and BFU-E-derived colonies.C-PC exhibited higher EPO-like activity.12.5 ng of C-PC could match with EPO 1.06U.The specific activity of C-PC was 84,800 U?mg~(-1) C-PC.When anemic mice were intra-peritoneal injected with C-PC for 5d,on d10 the leukocyte,erythrocyte and hemoglobin were significant increased.