1.Investigating the impact of silencing an RNA-binding protein gene SlRBP1 on tomato photosynthesis through RNA-sequencing analysis.
Xiwen ZHOU ; Liqun MA ; Hongliang ZHU
Chinese Journal of Biotechnology 2024;40(1):150-162
Photosynthesis in plants directly affects the synthesis and accumulation of organic matter, which directly influences crop yield. RNA-binding proteins (RBPs) are involved in the regulation of a variety of physiological functions in plants, while the functions of RBPs in photosynthesis have not been clearly elucidated. To investigate the effect of a glycine-rich RNA-binding protein (SlRBP1) in tomato on plant photosynthesis, a stably inherited SlRBP1 silenced plant in Alisa Craig was obtained by plant tissue culture using artificial small RNA interference. It turns out that the size of the tomato fruit was reduced and leaves significantly turned yellow. Chlorophyll(Chl) content measurement, Chl fluorescence imaging and chloroplast transmission electron microscopy revealed that the chloroplast morphology and structure of the leaves of tomato amiR-SlRBP1 silenced plants were disrupted, and the chlorophyll content was significantly reduced. Measurement of photosynthesis rate of wild-type and amiR-SlRBP1 silenced plants in the same period demonstrated that the photosynthetic rate of these plants was significantly reduced, and analysis of RNA-seq data indicated that silencing of SlRBP1 significantly reduced the expression of photosynthesis-related genes, such as PsaE, PsaL, and PsbY, and affected the yield of tomato fruits through photosynthesis.
RNA
;
Solanum lycopersicum/genetics*
;
Photosynthesis/genetics*
;
Chlorophyll
;
RNA-Binding Proteins/genetics*
2.Inhibition of Fibrotic Contraction by C-Phycocyanin through Modulation of Connective Tissue Growth Factor and α-Smooth Muscle Actin Expression.
Eunjin AN ; Hyunju PARK ; Ae Ri Cho LEE
Tissue Engineering and Regenerative Medicine 2016;13(4):388-395
The effects of C-phycocyanin (C-pc), a phycobiliprotein, on the expression of pro-fibrotic mediators in hyper-tropic scarring such as connective tissue growth factor (CTGF) and α-smooth muscle actins (α-SMA) were investigated in relation to trans-differentiation of fibroblast to myo-fibroblast, an icon of scar formation. C-pc was isolated from Spirulina Platensis extract using sonication method and C-pc concentration was determined by Bennet and Bogorad equation. α-SMA and CTGF levels in wounded primary human dermal fibroblasts were determined by western blot analysis and immuno-fluorescence confocal microscope was employed. Fibroblast contractility was examined by three-dimensional collagen lattice contraction assay. There was an elevation of α-SMA (121%) and CTGF (143%) levels in wound cells as compared with non-wound cells. The does-response profiles of down regulation demonstrated that the maximum inhibitions of α-SMA by 63% (p<0.05) and CTGF by 50% (p<0.1) were achieved by C-pc (6 nM) treated cells. In confocal assay, non-wound fibroblasts exhibited basal level of α-SMA staining, while wounded cells without C-pc treatment showed strong up-regulation of α-SMA by 147% (p<0.05). C-pc (6 nM) inhibited α-SMA expression by 70% (p<0.05) and reduced collagen contraction by 29% (p<0.05). C-pc seemed to lessen the over expression of CTGF, α-SMA, subsequently alleviating the fibrotic contracture. This study suggests the potential application of C-pc to regulation of the expression of pro-fibrotic mediators in scarring process and its potential usage as an efficient means for anti-fibrosis therapy.
Actins*
;
Blotting, Western
;
Cicatrix
;
Collagen
;
Connective Tissue Growth Factor*
;
Connective Tissue*
;
Contracture
;
Down-Regulation
;
Fibroblasts
;
Humans
;
Methods
;
Myofibroblasts
;
Phycocyanin*
;
Sonication
;
Spirulina
;
Up-Regulation
;
Wound Healing
;
Wounds and Injuries
3.Experimental study on the therapeutic effect of C-phycocyanin against pulmonary fibrosis induced by paraquat in rats.
Ying-xin SUN ; Juan ZHANG ; Gong-chang YU ; Yong-jian YAN ; Wen-wen CHEN ; Ming-feng CHI ; Song QIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(9):650-655
OBJECTIVETo investigate the therapeutic effect of C-phycocyanin (C-PC) from Spirulina platensis on paraquat (PQ)-induced pulmonary fibrosis in rats.
METHODSA total of 90 healthy Wistar rats were randomly and equally divided into normal control group, model group (PQ group), and C-PC treatment group (C-PC group). Each rat in the PQ group and C-PC group were orally administered with a single dose of PQ (50 mg/kg) to establish a rat model of PQ poisoning. Then, the rats in the normal control group and PQ group were orally given saline solution (1 ml/100 g) every day, and the rats in the C-PC group were orally given C-PC (50 mg/kg) every day. Six rats were randomly selected from each group on days 1, 3, 7, 14, and 28. The inferior lobe of each rat's right lung was homogenized for the measurement of hydroxyproline (HYP) and maleic dialdehyde (MDA) levels and superoxide dismutase (SOD) activity. Parts of each rat's left lung were subject to HE staining and Masson staining for pathological observation, and the expression of transforming growth factor-β(1) (TGF-β(1)), nuclear factor-kappa B p65 (NF-κB p65), and tumor necrosis factor-α (TNF-α) in lung tissue was measured by immunohistochemistry.
RESULTSThe HYP levels on days 1, 3, 7, 14, and 28 and MDA levels on days 14 and 28 were significantly lower in the C-PC group than in the PQ group (P < 0.05, P < 0.01). The SOD activity was significantly higher in the C-PC group than in the PQ group on days 1, 7, 14, and 28 (P < 0.05, P < 0.01). The protein content of TGF-β(1) and the activities of NF-κB p65 and TNF-α in the PQ group and C-PC group were significantly higher than those in the normal control group, while the indices in the C-PC group were significantly lower than those in the PQ group (P < 0.05, P < 0.01). The pathological observation showed that C-PC could alleviate pulmonary alveolitis and fibrosis in rats with PQ poisoning.
CONCLUSIONC-PC can significantly inhibit PQ-induced pulmonary alveolitis and fibrosis in rats.
Animals ; Lung ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Paraquat ; poisoning ; Phycocyanin ; pharmacology ; Pulmonary Fibrosis ; chemically induced ; metabolism ; prevention & control ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
4.Scale-up preparation of phycoerythrin from Porphyra haitanensis.
Chunxia LI ; Daiyuan YAN ; Jing NI ; Ziye GUO ; Chun'er CAI ; Peimin HE
Chinese Journal of Biotechnology 2011;27(4):614-619
We developed large-scale preparation of phycoerythrin from Porphyra haitanensis, a main economic red algae in China. Firstly, P. haitanensis thallus was broken by using "swelling and smash" method. Then times of grads ammonium sulfate precipitation applied to the crude extraction were compared. Desalted solution was further purified with one-step chromatography using hydroxyapatite and properties on spectrum and molecular weight were identified finally. The results indicated that after four times of ammonium sulfate precipitation (15%, 50%, 10% and 40%), the absorption spectrum purity of P. haitanensis achieved 0.9 (A564/A280), and 507.82 mg phycoerythrin (A564/A280 > 3.2) was obtained from 7 kg fresh algae after further hydroxyapatite chromatography. This research provides a potential way for preparation of phycoerythrin in large sclae.
Ammonium Sulfate
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chemistry
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Chromatography
;
methods
;
Phycoerythrin
;
isolation & purification
;
Porphyra
;
chemistry
5.Expression Pattern of the Thioredoxin System in Human Endothelial Progenitor Cells and Endothelial Cells Under Hypoxic Injury.
Keon Jae PARK ; Yeon Jeong KIM ; Eun Ju CHOI ; No Kwan PARK ; Gi Hyun KIM ; Sang Min KIM ; Sang Yeub LEE ; Jang Whan BAE ; Kyung Kuk HWANG ; Dong Woon KIM ; Myeong Chan CHO
Korean Circulation Journal 2010;40(12):651-658
BACKGROUND AND OBJECTIVES: The thioredoxin (TRx) system is a ubiquitous thiol oxidoreductase pathway that regulates cellular reduction/oxidation status. Although endothelial cell (EC) hypoxic damage is one of the important pathophysiologic mechanisms of ischemic heart disease, its relationship to the temporal expression pattern of the TRx system has not yet been elucidated well. The work presented here was performed to define the expression pattern of the TRx system and its correlation with cellular apoptosis in EC lines in hypoxic stress. These results should provide basic clues for applying aspects of the TRx system as a therapeutic molecule in cardiovascular diseases. SUBJECTS AND METHODS: Hypoxia was induced with 1% O2, generated in a BBL GasPak Pouch (Becton Dickinson, Franklin Lakes, NJ, USA) in human endothelial progenitor cells (hEPC) and human umbilical vein endothelial cells (HUVEC). Apoptosis of these cells was confirmed by Annexin-V: Phycoerythrin flow cytometry. Expression patterns of TRx; TRx reductase; TRx interacting protein; and survival signals, such as Bcl-2 and Bax, in ECs under hypoxia were checked. RESULTS: Apoptosis was evident after hypoxia in the two cell types. Higher TRx expression was observed at 12 hours after hypoxia in hEPCs and 12, 36, 72 hours of hypoxia in HUVECs. The expression patterns of the TRx system components showed correlation with EC apoptosis and cell survival markers. CONCLUSION: Hypoxia induced significant apoptosis and its related active changes of the TRx system were evident in human EC lines. If the cellular impact of TRx expression pattern in various cardiovascular tissues under hypoxia or oxidative stress was studied meticulously, the TRx system could be applied as a new therapeutic target in cardiovascular diseases, such as ischemic heart disease or atherosclerosis.
Anoxia
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Apoptosis
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Atherosclerosis
;
Cardiovascular Diseases
;
Cell Hypoxia
;
Cell Survival
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Endothelial Cells
;
Flow Cytometry
;
Human Umbilical Vein Endothelial Cells
;
Humans
;
Lakes
;
Myocardial Ischemia
;
Oxidative Stress
;
Phycoerythrin
;
Stem Cells
;
Thioredoxins
6.C-phycocyanin Attenuates Renal Inflammation and Fibrosis in UUO Mice.
Sarah CHUNG ; Jin Young JEONG ; Dae Eun CHOI ; Ki Ryang NA ; Kang Wook LEE ; Young Tai SHIN
Korean Journal of Nephrology 2010;29(6):687-694
PURPOSE: It has been reported that Spirulina, a blue-green algae with potent antioxidant properties, affords significant protection against inflammation and fibrosis in the liver in vivo. The aim of the present study was to establish the possible protective role of C-phycocyanin, one of the active ingredients of Spirulina, in an experimental model of fibrosis in the kidney. METHODS: The study was carried out using male C57BL6 mice. Mice were divided into the following four groups: sham-operated group; C-phycocyanin (PC)-treated sham group; unilateral ureteral obstruction (UUO) group; and PC with UUO group. We evaluated renal TGF-beta mRNA, MCP-1, and osteopontin using real-time RT PCR. We evaluated renal TGF-beta, alpha-SMA, and CD68 by immunohistochemistry. We recorded light microscopic findings of kidney specimens. RESULTS: PC significantly decreased the expression of MCP-1 and alpha-SMA mRNA. Renal gene levels of expression of TGF-beta, MCP-1, and osteopontin in the UUO group were significantly higher than the sham-operated group (p<0.01). The levels of expression of TGF-beta, MCP-1, and osteopontin mRNA of kidneys in the PC-treated UUO group were significantly lower than the untreated UUO group (p< 0.05). The magnitude of expression of TGF-beta and alpha-SMA protein in the kidneys of the PC-treated UUO group was significantly less than the untreated UUO control group (p<0.05). CONCLUSION: The results of the present study suggest that PC has anti-inflammatory and anti-fibrotic effects in an experimental UUO murine model.
Animals
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Cyanobacteria
;
Fibrosis
;
Humans
;
Immunohistochemistry
;
Inflammation
;
Kidney
;
Light
;
Liver
;
Male
;
Mice
;
Models, Theoretical
;
Osteopontin
;
Phycocyanin
;
Polymerase Chain Reaction
;
RNA, Messenger
;
Salicylamides
;
Spirulina
;
Transforming Growth Factor beta
;
Ureteral Obstruction
7.Protective Effect of C-phycocyanin against Inflammation and Interstitial Fibrosis in an Obstructive Nephropathy Model.
Korean Journal of Nephrology 2010;29(6):683-686
No abstract available.
Fibrosis
;
Inflammation
;
Phycocyanin
9.Photodynamic effect of two kinds of phycobiliproteins on human liver cancer cell line SMMC-7721 in vitro.
Yuan WANG ; Chuner CAI ; Bailin LI ; Chengchu LIU ; Peimin HE
Chinese Journal of Biotechnology 2009;25(9):1417-1423
We studied the effect of photodynamic therapy with phycobiliproteins on human liver cancer cells in vitro. With 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT assay), we used two phycobiliproteins, R-phycoerythrin (R-PE) and C-phycocyanin (C-PC) prepared from Porphyra yezoensis, to determine the killing rates and apoptosis rates of human liver cancer cells (SMMC-7721) mediated by laser. When the concentration of R-PE was 120 mg/L, the survival rate of human liver cancer cells was 27% after treated by Argon laser with 100 J/cm2 doses, while the survival rate in the control group (without adding R-PE) was 65%. When the C-PC concentration was 120 mg/L, the survival cell rate was 47% after treated by He-Ne laser with 35 J/cm2 dose, while the survival rate in the control group (without adding C-PC) was 70%. After handled only with these two kinds of phycobiliproteins for 72 h, the growth of cancer cells presented significant inhibition. The maximal inhibition rates reached up to 31% with R-PE (120 mg/L concentration) and 27% with C-PC (250 mg/L concentration) respectively. After irradiated by laser for 8 h, the maximal cell apoptosis rates were 31.54% with R-PE and 32.54% with C-PC, respectively. It indicated that R-PE and C-PC extracted from Porphyra yezoensis could develop to new photosensitizers for cancer photodynamic therapy.
Apoptosis
;
drug effects
;
radiation effects
;
Cell Line, Tumor
;
Humans
;
Lasers
;
Liver Neoplasms
;
pathology
;
Photochemical Processes
;
Photochemotherapy
;
methods
;
Phycobiliproteins
;
isolation & purification
;
pharmacology
;
Phycoerythrin
;
isolation & purification
;
pharmacology
;
Porphyra
;
chemistry
10.Preparation of phycocyanin subunits liposomes and the photodynamic experiment on cancer cells.
Rui-yong GUO ; Bei HUANG ; Man-man ZUO ; Yong-zhong WANG ; Ling HU
Acta Pharmaceutica Sinica 2008;43(10):1060-1065
Phycocyanin subunits liposomes (PCS-lip) were prepared and its cellular uptake and photodynamic therapy (PDT) effect on cancer cells were studied. In the experiment, film dispersion method was used to prepare phycocyanin subunits liposomes; particle size and distribution were detected by zetasizer and transmission electric microscope; the effects of liposome as carrier on cell uptake in vitro were evaluated in S180 by using fluorescence microscope; and photodynamic therapy effect was assessed with MTT method. As shown in the results, the particle size mainly ranged from 80 nm to 160 nm, and average encapsulation rate was 42.3%. In the concentration of 100 microg x mL(-1), transfection rate reached (18.5 +/- 0.8)% at 2 h, (23.1 +/- 0.9)% at 4 h, keeping a balance in 5-6 h, and its photodynamic therapy effect in vitro improved with the increasing of concentration of phycocyanin subunits liposomes. In the concentration of 200 micro x mL(-1) cell survival rate of BGC-823 and S180 reached (45 +/- 5.2)% and (36 +/- 5.5)%, respectively, and the cell survival rate differentiation between PCS-PDT group and PCS-lip-PDT group reached 7%-11% (P < 0.05). In this study film dispersion method could keep the biological activity of phycocyanin subunits very well. Phycocyanin subunits liposomes will transfect cells more quickly than phycocyanin subunits in the same concentration, and in the same conditions, phycocyanin subunits liposomes have the better PDT effect on cancer cells as they were incubated with cells for 4 h.
Animals
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Cell Line, Tumor
;
Cell Survival
;
drug effects
;
Drug Carriers
;
Drug Stability
;
Humans
;
Liposomes
;
chemistry
;
Mice
;
Particle Size
;
Photochemotherapy
;
methods
;
Phycocyanin
;
administration & dosage
;
pharmacology
;
Protein Subunits
;
administration & dosage
;
pharmacology
;
Sarcoma 180
;
pathology
;
Stomach Neoplasms
;
pathology

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