Photoreceptor cell degeneration leads to blindness, for which there is currently no effective treatment. Our previous studies have shown that Lycium barbarum(L. barbarum) polysaccharide(LBP) protects degenerated photo...Photoreceptor cell degeneration leads to blindness, for which there is currently no effective treatment. Our previous studies have shown that Lycium barbarum(L. barbarum) polysaccharide(LBP) protects degenerated photoreceptors in rd1, a transgenic mouse model of retinitis pigmentosa. L. barbarum glycopeptide(Lb GP) is an immunoreactive glycoprotein extracted from LBP. In this study, we investigated the potential protective effect of Lb GP on a chemically induced photoreceptor-degenerative mouse model. Wild-type mice received the following: oral administration of Lb GP as a protective pre-treatment on days 1–7;intraperitoneal administration of 40 mg/kg N-methylN-nitrosourea to induce photoreceptor injury on day 7;and continuation of orally administered Lb GP on days 8–14. Treatment with Lb GP increased photoreceptor survival and improved the structure of photoreceptors, retinal photoresponse, and visual behaviors of mice with photoreceptor degeneration. Lb GP was also found to partially inhibit the activation of microglia in N-methyl-N-nitrosourea-injured retinas and significantly decreased the expression of two pro-inflammatory cytokines. In conclusion, Lb GP effectively slowed the rate of photoreceptor degeneration in N-methyl-N-nitrosourea-injured mice, possibly through an anti-inflammatory mechanism, and has potential as a candidate drug for the clinical treatment of photoreceptor degeneration.展开更多
AIM:To investigate the antioxidant protective effect of Lycium barbarum glycopeptide(LbGP)pretreatment on retinal ischemia-reperfusion(I/R)injury(RIRI)in rats.METHODS:RIRI was induced in Sprague Dawley rats through an...AIM:To investigate the antioxidant protective effect of Lycium barbarum glycopeptide(LbGP)pretreatment on retinal ischemia-reperfusion(I/R)injury(RIRI)in rats.METHODS:RIRI was induced in Sprague Dawley rats through anterior chamber perfusion,and pretreatment involved administering LbGP via gavage for 7d.After 24h of reperfusion,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and creatinine(CREA)levels,retinal structure,expression of Caspase-3 and Caspase-8,superoxide dismutase(SOD)activity,and malondialdehyde(MDA)in the retina were measured.RESULTS:The pretreatment with LbGP effectively protected the retina and retinal tissue from edema and inflammation in the ganglion cell layer(GCL)and nerve fiber layer(NFL)of rats subjected to RIRI,as shown by light microscopy and optical coherence tomography(OCT).Serum AST was higher in the model group than in the blank group(P=0.042),but no difference was found in ALT,AST,and CREA across the LbGP groups and model group.Caspase-3 expression was higher in the model group than in the blank group(P=0.006),but no difference was found among LbGP groups and the model group.Caspase-8 expression was higher in the model group than in the blank group(P=0.000),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).SOD activity was lower in the model group than in the blank group(P=0.001),and the decrease was slower in the 400 mg/kg LbGP group than in the model group(P=0.003).MDA content was higher in the model group than in the blank group(P=0.001),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).The pretreatment with LbGP did not result in any observed liver or renal toxicity in the model.CONCLUSION:LbGP pretreatment exhibits dosedependent anti-inflammatory,and antioxidative effects by reducing Caspase-8 expression,preventing declines of SOD activity,and decreasing MDA content in the RIRI rat model.展开更多
Objective To study whether Lycium barbarurn glycopeplide 3 （LBGP3） affects T cell apoptosis in aged mice. Methods LBGP3 was purified with DEAE cellulose and Sephadex columns. Apoptotic ＂sub-G1 peak＂ was detected b...Objective To study whether Lycium barbarurn glycopeplide 3 （LBGP3） affects T cell apoptosis in aged mice. Methods LBGP3 was purified with DEAE cellulose and Sephadex columns. Apoptotic ＂sub-G1 peak＂ was detected by flow cytometry and DNA ladder was resolved by agarose gel electrophoresis. Levels of IFN-7 and IL-10 were measured with specific kits and mRNA expression was detected by RT-PCR. Apoptosis-related proteins of FLIP, FasL, and Bcl-2 were determined by Western blotting. Results LBGP3 was purified from Fructus Lycii water extracts and identified as a 41 kD glycopeptide. Treatment with 200 p.g/mL LBGP3 increased the apoptotic rate of T cells from aged mice and showed a similar DNA ladder pattern to that in young T cells. The reversal of apoptotic resistance was involved in down-regulating the expression of Bcl-2 and FLIP, and up-regulating the expression of FasL. Conclusion Lycium barbarum glycopeptide 3 reverses apoptotic resistance of aged T cells by modulating the expression of apoptosis-related molecules.展开更多
In order to investigate the immunoactivity of Lycium Barbarum glycopeptide (LBG), the routinely prepared murine splenic lymphocyte suspension was separately added into the samples with different concentrations (500, 1...In order to investigate the immunoactivity of Lycium Barbarum glycopeptide (LBG), the routinely prepared murine splenic lymphocyte suspension was separately added into the samples with different concentrations (500, 100, 10, 1 μg/ml) of LBG as LBG groups. Blank control group in the absence of Lycium Barbarum glycopeptide or ConA and positive control group in the presence of 0.5 ml ConA but in the absence of LBG were created. 0.5 ml LBG samples with different concentrations in combination with 0.5 ml ConA (10 μg/ml) into each well to observe the synergic effects of LBG and ConA as LBG+ConA groups. After incubation for 72 h at 37 ℃, the samples were analyzed by CFSE-labeled cells combined with flow cytometry, and MTT. Flow cytometry revealed that both LBG could enhance the murine splenic lymphocyte proliferative reaction. Combined use of LBG and ConA had synergic effects. MTT demonstrated that sample A could obviously promote the murine splenic lymphocyte proliferative reaction as compared with control group (P<0.01), while sample B could also enhance the lymphocyte proliferation at a high dose. In combination with ConA, sample A had synergic effects at high dose, while sample B showed obviously synergic effects (P<0.05). It was concluded that both samples (A and B) had strong immunocompetence.展开更多
Investigations of glycosylated proteins or peptides and their related biological pathways provide new possibilities for illuminating the physiological and pathological mechanisms of glycosylation modification.However,...Investigations of glycosylated proteins or peptides and their related biological pathways provide new possibilities for illuminating the physiological and pathological mechanisms of glycosylation modification.However,open-ended and in-depth analysis of glycoproteomics is usually subjected to the low-abundance of glycopeptides,heterogeneous glycans,and a variety of interference molecules.In order to alleviate the influence of these obstacles,effective preconcentration of glycopeptides are indispensable.Here,we employed a hydrophilic interaction liquid chromatography(HILIC)-based method to universally capture glycopeptides.Glutathione modified magnetic nanoparticles(Fe304@AuGSH) were synthesized through a simple process and exploited to enrich glycopeptides from complex samples.The prepared materials showed excellent ability to trap glycopeptides from standard glycoproteins digests,low detection limit(10 fmol/p L),and good selectivity(HRP:BSA=1:100).These results indicated that glutathione-based magnetic nanoparticles synthesized in this work had great potential for glycopeptides enrichment.展开更多
This study was conducted to recover edible bird’s nest(EBN)hydrolysates from different grades of EBN,including the industrial by-products,using enzymatic treatment.The nutrient,physicochemical properties and antioxid...This study was conducted to recover edible bird’s nest(EBN)hydrolysates from different grades of EBN,including the industrial by-products,using enzymatic treatment.The nutrient,physicochemical properties and antioxidant activities of the recovered hydrolysates at different hydrolysis times were evaluated.Results showed that the recovery yield of enzymatic hydrolysis was above 89%for all grades of EBN and the degree of hydrolysis increased over time.Nitrite content(0.321-0.433 mg/L)was below the permissible tolerance level for all samples.Interestingly,the antioxidant activities(DPPH and ABTS scavenging activities and ferric reducing antioxidant powder(FRAP)activity)were significantly higher(P≤0.05)in hydrolysates recovered from EBN by-products(EBNhC and EBNhD)as compared to the high grade EBN hydrolysates(EBNhA and EBNhB).The in-vitro probiotic activity of EBN and its hydrolysates were examined using the probiotic bacterium Lactobacillus plantarum.Evidently,EBN by-products hydrolysate(EBNhD)recorded the highest number of L.plantarum(1.1×1011 CFU/mL),indicating that low grade EBN has the potential as prebiotic material that promotes probiotic activity.This study demonstrated the concept of using EBN by-products hydrolysates for various applications,such as functional ingredients with enhanced bioactivities,to improve its economic value.展开更多
Our previous reports have shown that lamininglycopeptides (LN-GPs), the total glycopeptides prepared from laminin (LN), can prevent the experimental lung metastasis and liver metastasis of mouse cancer cells. In order...Our previous reports have shown that lamininglycopeptides (LN-GPs), the total glycopeptides prepared from laminin (LN), can prevent the experimental lung metastasis and liver metastasis of mouse cancer cells. In order to explore the anti-metastatic mechanism of LNGPs, we studied the effects of LN-GPs on metastasisrelated behaviors of cancer cells in vitro. LN-GPs did not affect cell survival. However, LN-GPs inhibited cell attachment and spreading Of 5180 cells on LN- and Matrigelsubstrate in dose-dependent and time-dependent manners. Moreover, inhibition of cen attachment and spreading on Matrigel substrates were much greater on Matrigel substrate than on LN substrate. In the presence of LN-GPs, 5180 cells on LN substrate changed from a flattened polygonal shape to a round one, the migration of 5180 cells on LN substrate decreased, and the number of a highly invasive human pulmonary giant caxcinoma PG cells invading Matrigel filter in a Boyden chamber was reduced. LN-GPs thus have multiple inhibitory effects on cancer motastasisrelated behaviors.展开更多
The analysis of endogenous glycoproteins and glycopeptides in human body fluids is of great importance for screening and discovering disease biomarkers with clinical significance.However,the presence of interfering su...The analysis of endogenous glycoproteins and glycopeptides in human body fluids is of great importance for screening and discovering disease biomarkers with clinical significance.However,the presence of interfering substances makes the direct quantitative detection of low-abundance glycoproteins and glycopeptides in human body fluids one of the great challenges in analytical chemistry.Magnetic solid phase extraction(MSPE)has the advantages of easy preparation,low cost and good magnetic responsiveness.Magnetic adsorbents are the core of MSPE technology,and magnetic adsorbents based on different functional materials are widely used in the quantitative analysis of glycoproteins and glycopeptides in human body fluids,making it possible to analyze glycoproteins and glycopeptides with low abundance as well as multiple types,which provides a technical platform for screening and evaluating glycoproteins and glycopeptides in body fluids as disease biomarkers.In this paper,we focus on the recent advances in the application of MSPE technology and magnetic adsorbents for the separation and enrichment of glycoproteins and glycopeptides in human body fluids,and the future trends and application prospects in this field are also presented.展开更多
Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor ...Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor selectivity. The present study is the first attempt to combine chitosan(CS) and L-cysteine(L-Cys) to design a novel hydrophilic material focusing on glycopeptide enrichment. CS containing a large number of hydrophilic amino and hydroxyl groups has unique chemical properties, which makes it a very attractive biomaterial for glycopeptide enrichment. The excellent hydrophilicity of zwitterionic molecule L-Cys inspires the idea of anchoring L-Cys onto CS to design a novel hydrophilic material(named as Fe_(3)O_(4)@CS@Au-L-Cys) for the capture of low abundance glycopeptides. To be specific, Au nanoparticles(Au NPs) was introduced into CS-coated Fe_(3)O_(4)via electrostatic interaction and served as bridges to anchor L-Cys onto the surface of CS through strong Au-S bond interaction. The prepared Fe_(3)O_(4)@CS@AuL-Cys exhibited strong affinity, low detection limit(0.5 fmol/μL HRP), high selectivity(HRP/BSA with a molar ratio of 1:1000) for glycopeptides. Moreover, successful application of glycopeptide enrichment in human serum and saliva by Fe_(3)O_(4)@CS@Au-L-Cys was achieved. A satisfactory data set indicates that Fe_(3)O_(4)@CS@Au-L-Cys has promising potential in the application of glycopeptide enrichment in real complex bio-samples and for related glycoproteome research.展开更多
A mixture of oligosaccharide and glycopeptide was isolated from the aqueous extract of Panax ginseng roots. The mixture inhibits protein tyrosinc phosphatase（SHP-1） function, implying it enhances immune activity. Th...A mixture of oligosaccharide and glycopeptide was isolated from the aqueous extract of Panax ginseng roots. The mixture inhibits protein tyrosinc phosphatase（SHP-1） function, implying it enhances immune activity. The peak molecular mass of the oligosaccharide portion is 1800 calculated via GPC software after separation by HPLC. And the structure of the oligosaccharide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→4）- and （1→6）-linked glucopyranoside, with non-reducing terminals of arabinopyranoside and glucopyranoside. The peak molecular mass of glycopeptide portion is 1900 calculated via GPC software after separation by HPLC. The structure of glycopeptide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→3,6）-linked glucopyranoside, with non-reducing terminals of galactopyranose and glucopyranoside. The peptide composition is Glu. Asp, Hyp, Set, Arg, Gly , Thr, Pro, Ala, Val, lie, Leu and Lys. The oligosaccharide-peptide linkage is formed by Ara and Hyp.展开更多
Exosome and inclusive cargoes have manifested significant function in different biological events. In particular, glycopeptides in exosome are closely associated with occurrence and development of various diseases. De...Exosome and inclusive cargoes have manifested significant function in different biological events. In particular, glycopeptides in exosome are closely associated with occurrence and development of various diseases. Developing advanced tools is highly desired to enrich glycopeptides from exosomes, and enrich exosomes from complex biological samples as well. In this work, integration of L-cysteine and titania onto the surface of magnetic nanoparticles is designed to realize the coefficient affinity towards exosomes and inclusive glycopeptides. Benefiting from the synergistic affinity, we separate exosomes from human urine concentrate directly, which was proved by the detection of three typical antigen markers of exosomes. Furthermore, hardly any exosomes remained on materials after ultrasonication, which confirmed the good capture performance of Fe_(3)O_(4)@TiO_(2)@L-Cys and high release effect of direct lysis.Moreover, 146 glycopeptides corresponding to 77 glycoproteins were successfully identified from captured exosomes. These satisfactory results will inspire more efforts to be devoted to this field and will be extremely helpful to in-depth information excavation of biological markers, especially disease-related ones, through exosomes and inclusive glycopeptides.展开更多
β-Cyclodextrin(β-CD) based materials have attracted great attention in the separation of hydrophilic glycopeptides due to the abundant hydroxyl groups in its exterior. However, the current materials based on β-CD g...β-Cyclodextrin(β-CD) based materials have attracted great attention in the separation of hydrophilic glycopeptides due to the abundant hydroxyl groups in its exterior. However, the current materials based on β-CD generally has complex synthesis process and harsh experimental conditions, on the other hand,the interior cavity of β-CD is hydrophobic and is harmful to capture glycopeptides. Herein, a novel hydrophilic material based on β-CD was engineered via a self-assembly process utilizing L-cysteine(L-Cys)or glutathione(GSH) derived adamantane for highly efficient glycopeptide enrichment. It is the first attempt to make use of the hydrophobic interior cavity of β-CD for hydrophilic glycopeptide capture. Taking advantages of strong hydrophilicity and superparamagnetism, the as-prepared materials possess low detection limit, high selectively, and excellent reusability when employed to glycopeptide enrichment. In addition, the feasibility of the hydrophilic material based on β-CD was verified by enriching glycopeptides from human serum and saliva samples. This study provides a heuristic strategy for the application of β-CD-based self-assembly materials in the enrichment of glycopeptides. Importantly, this strategy certified a possible that the change of glycopeptide enrichment sites through host-vip interaction between β-CD and adamantane derivatives with different functional groups.展开更多
Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction m...Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction methods,namely,water extraction,ultrasonic extraction,enzymatic hydrolysis-water extraction,and enzymatic hydrolysis-ultrasonic extraction were considered to determine the best extraction method.Single factor and orthogonal experiments were performed to determine the optimum extracting conditions.Sevage,enzymatic hydrolysis-Sevage,trichloroacetic acid(TCA),TCA-Sevage and enzymatic hydrolysis-TCA methods were tested to determine the best deproteinization method.The glycopeptide content and protein removal ratio were analyzed by the phenol-sulfuric acid and Coomassie Brilliant Blue methods.Results:Gekko sulfated glycopeptide obtained by water extraction could inhibit the proliferation of HepG2 and SKBR3,as well as promote that of lymphocytes.The glycopeptide content was 4.049%in the optimal extracting condition of a triple decoction extraction for 1 hour each time with a material to solvent ratio of 1:15.The enzymatic hydrolysis-TCA method was found to be the optimal deproteinization method,with a protein removal ratio of 50.46%,glycopeptide content of 14.27%,and inhibitory ratio on human HepG2 cells of 49.06%.Conclusion:This extraction and purification technique for Gekko sulfated glycopeptide is reasonable,feasible,and provides a scientific basis for industrial production.展开更多
Both glycosylation and phosphorylation exert crucial rule in multitudinous biological processes.For in-depth profiling of glycosylation and phosphorylation,a magnetic metal oxide is effectively coupled with inherently...Both glycosylation and phosphorylation exert crucial rule in multitudinous biological processes.For in-depth profiling of glycosylation and phosphorylation,a magnetic metal oxide is effectively coupled with inherently hydrophilic mesoporous channels(denoted as Fe_(3)O_(4)@TiO_(2)@mSiO_(2)-TSG).Based on the mechanism of hydrophilic interaction liquid chromatography(HILIC)and metal oxide affinity chromatography(MOAC),the Fe_(3)O_(4)@TiO_(2)@mSiO_(2)-TSG nanomaterial shows high capacity for simultaneously enriching glycopeptides and phosphopeptides.With human saliva collected in successive four days as practical biological sample,endogenous glycopeptides and phosphopeptides are efficiently enriched.Further gene ontology analysis reveals that the identified endogenous glycopeptides and phosphopeptides participate in diverse molecular functions and biological processes.This strategy is anticipated to promote variation analysis of salivary post-translational modifications.展开更多
The detailed glycan structural analysis of glycoprotein is amenable to glycopeptide enrichment. Here, we develop a simple, effective and economical approach to enrich glycopeptides from proteolytically digested peptid...The detailed glycan structural analysis of glycoprotein is amenable to glycopeptide enrichment. Here, we develop a simple, effective and economical approach to enrich glycopeptides from proteolytically digested peptide mixtures by chromatographic column packed with graphite carbon and activated charcoal (G/A-column). Glycopeptide from ovalbumin was efficiently enriched by homemade G/A-column using liquid chromatography and the structure of glycopeptide was obtained by tandem mass spectrometry using Fourier transform ion cyclotron resonance mass spectrometry. The results in this study demonstrate that G/A-column can be used to enrich N-glycolpeptides and be benefit for online identification of glycopeptide using LC-MS.展开更多
There are also a variety of cytokines in the tumor microenvironment,which are involved in the change in the hardness of the tumor,thereby affecting the invasion and metastasis of tumor cells.As traditional Chinese med...There are also a variety of cytokines in the tumor microenvironment,which are involved in the change in the hardness of the tumor,thereby affecting the invasion and metastasis of tumor cells.As traditional Chinese medicines,drugs of softening and dissipating firm knot contains different kinds of sulfated polysaccharide and sulfated glycopeptide.By inhibiting the function of cancer-associated fibroblasts,they reduce interstitial fibrosis,thereby reducing the hardness of the tumor and exerting an anti-tumor effect.展开更多
Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting b...Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting basic fibroblast growth factor(bFGF).bFGF promoted the proliferation of fibroblasts.Both fibrosarcoma and fibroblasts derived from fibrous connective tissue.This study investigated whether GSPP has the inhibitory effects on human fibrosarcoma HT1080 cells.Materials and methods:The trypan blue exclusion assay was used to determine cell viability and cell numbers.Cells migration was observed by wound-healing and transwell.Results:From the first day to seventh day,HT1080 cells number of GSPP,bFGF,GSPP combined bFGF groups had not change compared with control.HT1080 cells migration distance and the number of migrating cells of GSPP,bFGF,GSPP combined bFGF groups were not significantly reduced.Conclusions:GSPP did not have inhibitory effects on the proliferation and migration of human fibrosarcoma HT1080 cells.Thus further research should be carried out in order to study the mechanism of GSPP and bFGF acting on the tumor stroma.展开更多
Background:Our previous work demonstrated that Gekko sulfated glycopeptide extracted from the Chinese gecko Shou gong(Gekko swinhonis Guenther)could inhibit tumor growth by regulating basic fibroblast growth factor.Ho...Background:Our previous work demonstrated that Gekko sulfated glycopeptide extracted from the Chinese gecko Shou gong(Gekko swinhonis Guenther)could inhibit tumor growth by regulating basic fibroblast growth factor.However,basic fibroblast growth factor has opposing effects on growth in breast and liver cancers.Direct effects and mechanisms of Gekko sulfated glycopeptide on tumor growth remain unclear and are ripe for further exploration.Methods:Differential regulation by Gekko sulfated glycopeptide and bFGF were studied in established human breast cancer MCF-7 cells and hepatocarcinoma HepG2 cells.Cell proliferation was evaluated with a Trypan blue exclusion assay.Cell cycle phases were measured by flow cytometry.Basic fibroblast growth factor,transforming growth factor-β1,and p21WAF1/CIP1 mRNA and protein expression levels were detected by real-time PCR(mRNA)and ELISA(protein).Changes in phosphorylated extracellular signal-regulated kinase levels were analyzed by Western blot.Results:Data indicated that Gekko sulfated glycopeptide inhibited the proliferation of HepG2 cells(P<0.001)and also blocked basic fibroblast growth factor-stimulated proliferation of these cells(P=0.001).Gekko sulfated glycopeptide was shown to increase transforming growth factor-β1 and p21WAF1/CiP1 expression(P<0.01)and partially compensate for reductions therein induced by basic fibroblast growth factor.Conversely,in MCF-7 cells,Gekko sulfated glycopeptide alone had no observable effect on transforming growth factor-β1 and p21WAF1/CiP1 expression.Gekko sulfated glycopeptide did,however,enhance basic fibroblast growth factor-induced inhibition of cell proliferation and transforming growth factor-β1 and p21WAF1/CiP1 expression in the MCF-7 cells(P=0.001,P<0.01,P<0.01,respectively).Gekko sulfated glycopeptide was shown to suppress basic fibroblast growth factor secretion in both HepG2 and MCF-7 cell lines(P<0.05 and P<0.01,respectively)and inhibited extracellular signal-regulated kinase 1/2 phosphorylation facilitated by basic fibroblast growth factor.Gekko sulfated glycopeptide alone decreased phosphorylated extracellular signal-regulated kinase in HepG2 cells but did not visibly affect phosphorylated extracellular signal-regulated kinase levels in MCF-7 cells.Conclusions:Gekko sulfated glycopeptide,a basic fibroblast growth factor inhibitor,differentially regulates growth in different cancer cell lines,and these differences may be determined by the opposing effects of basic fibroblast growth factor on transforming growth factor-β1 and p21WAF1/CiP1 levels in breast and liver cancer cells.展开更多
Cartilage injury represents a frequent dilemma in clinical practice owing to its inherently limited self-renewal capacity.Biomimetic strategy-based engineered biomaterial,capable of coordinated regulation for cellular...Cartilage injury represents a frequent dilemma in clinical practice owing to its inherently limited self-renewal capacity.Biomimetic strategy-based engineered biomaterial,capable of coordinated regulation for cellular and microenvironmental crosstalk,provides an adequate avenue to boost cartilage regeneration.The level of oxidative stress in microenvironments is verified to be vital for tissue regeneration,yet it is often overlooked in engineered biomaterials for cartilage regeneration.Herein,inspired by natural cartilage architecture,a fibril-network glycopeptide hydrogel(Nap-FFGRGD@FU),composed of marine-derived polysaccharide fucoidan(FU)and naphthalenephenylalanine-phenylalanine-glycine-arginine-glycine-aspartic peptide(Nap-FFGRGD),was presented through a simple supramolecular self-assembly approach.The Nap-FFGRGD@FU hydrogels exhibit a native cartilage-like architecture,characterized by interwoven collagen fibers and attached proteoglycans.Beyond structural simulation,fucoidan-exerted robust biological effects and Arg-Gly-Asp(RGD)sequence-provided cell attachment sites realized functional reinforcement,synergistically promoted extracellular matrix(ECM)production and reactive oxygen species(ROS)elimination,thus contributing to chondrocytes-ECM harmony.In vitro co-culture with glycopeptide hydrogels not only facilitated cartilage ECM anabolic metabolism but also scavenged ROS accumulation in chondrocytes.Mechanistically,the chondro-protective effects induced by glycopeptide hydrogels rely on the activation of endogenous antioxidant pathways associated with nuclear factor erythroid 2-related factor 2(NRF2).In vivo implantation of glycopeptide hydrogels successfully improved the de novo cartilage generation by 1.65-fold,concomitant with coordinately restructured subchondral bone structure.Collectively,our ingeniously crafted bionic glycopeptide hydrogels simultaneously rewired chondrocytes’function by augmenting anabolic metabolism and rebuilt ECM microenvironment via preserving redox equilibrium,holding great potential for cartilage tissue engineering.展开更多
基金supported by Guangzhou Key Projects of Brain Science and Brain-Like Intelligence Technology,No.20200730009 (to YX)the National Natural Science Foundation of China,No.82074169 (to XM)+2 种基金the Guangdong Basic and Applied Basic Research Foundation,No.2021A1515012473 (to XM)Project of Administration of Traditional Chinese Medicine of Guangdong Province,No.20202045 (to XM)Aier Eye Hospital Group,No.AF2019001 (to ST,KFS,YX,XM)。
文摘Photoreceptor cell degeneration leads to blindness, for which there is currently no effective treatment. Our previous studies have shown that Lycium barbarum(L. barbarum) polysaccharide(LBP) protects degenerated photoreceptors in rd1, a transgenic mouse model of retinitis pigmentosa. L. barbarum glycopeptide(Lb GP) is an immunoreactive glycoprotein extracted from LBP. In this study, we investigated the potential protective effect of Lb GP on a chemically induced photoreceptor-degenerative mouse model. Wild-type mice received the following: oral administration of Lb GP as a protective pre-treatment on days 1–7;intraperitoneal administration of 40 mg/kg N-methylN-nitrosourea to induce photoreceptor injury on day 7;and continuation of orally administered Lb GP on days 8–14. Treatment with Lb GP increased photoreceptor survival and improved the structure of photoreceptors, retinal photoresponse, and visual behaviors of mice with photoreceptor degeneration. Lb GP was also found to partially inhibit the activation of microglia in N-methyl-N-nitrosourea-injured retinas and significantly decreased the expression of two pro-inflammatory cytokines. In conclusion, Lb GP effectively slowed the rate of photoreceptor degeneration in N-methyl-N-nitrosourea-injured mice, possibly through an anti-inflammatory mechanism, and has potential as a candidate drug for the clinical treatment of photoreceptor degeneration.
基金Supported by the National Natural Science Foundation of China(No.82174444)the Chengdu University of Traditional Chinese Medicine Xinglin Scholar Discipline Talent Research Promotion Program Project(No.XKTD2022009)the Inheritance and Communication Department of Science and Technology Innovation Engineering Department of Chinese Academy of Chinese Medical Sciences(No.XJ2023001701).
文摘AIM:To investigate the antioxidant protective effect of Lycium barbarum glycopeptide(LbGP)pretreatment on retinal ischemia-reperfusion(I/R)injury(RIRI)in rats.METHODS:RIRI was induced in Sprague Dawley rats through anterior chamber perfusion,and pretreatment involved administering LbGP via gavage for 7d.After 24h of reperfusion,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and creatinine(CREA)levels,retinal structure,expression of Caspase-3 and Caspase-8,superoxide dismutase(SOD)activity,and malondialdehyde(MDA)in the retina were measured.RESULTS:The pretreatment with LbGP effectively protected the retina and retinal tissue from edema and inflammation in the ganglion cell layer(GCL)and nerve fiber layer(NFL)of rats subjected to RIRI,as shown by light microscopy and optical coherence tomography(OCT).Serum AST was higher in the model group than in the blank group(P=0.042),but no difference was found in ALT,AST,and CREA across the LbGP groups and model group.Caspase-3 expression was higher in the model group than in the blank group(P=0.006),but no difference was found among LbGP groups and the model group.Caspase-8 expression was higher in the model group than in the blank group(P=0.000),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).SOD activity was lower in the model group than in the blank group(P=0.001),and the decrease was slower in the 400 mg/kg LbGP group than in the model group(P=0.003).MDA content was higher in the model group than in the blank group(P=0.001),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).The pretreatment with LbGP did not result in any observed liver or renal toxicity in the model.CONCLUSION:LbGP pretreatment exhibits dosedependent anti-inflammatory,and antioxidative effects by reducing Caspase-8 expression,preventing declines of SOD activity,and decreasing MDA content in the RIRI rat model.
基金This research was supported by the National Basic Research Program of China (No. 2007CB507406) National Natural Science Foundation of China (No. 30600659)
文摘Objective To study whether Lycium barbarurn glycopeplide 3 （LBGP3） affects T cell apoptosis in aged mice. Methods LBGP3 was purified with DEAE cellulose and Sephadex columns. Apoptotic ＂sub-G1 peak＂ was detected by flow cytometry and DNA ladder was resolved by agarose gel electrophoresis. Levels of IFN-7 and IL-10 were measured with specific kits and mRNA expression was detected by RT-PCR. Apoptosis-related proteins of FLIP, FasL, and Bcl-2 were determined by Western blotting. Results LBGP3 was purified from Fructus Lycii water extracts and identified as a 41 kD glycopeptide. Treatment with 200 p.g/mL LBGP3 increased the apoptotic rate of T cells from aged mice and showed a similar DNA ladder pattern to that in young T cells. The reversal of apoptotic resistance was involved in down-regulating the expression of Bcl-2 and FLIP, and up-regulating the expression of FasL. Conclusion Lycium barbarum glycopeptide 3 reverses apoptotic resistance of aged T cells by modulating the expression of apoptosis-related molecules.
文摘In order to investigate the immunoactivity of Lycium Barbarum glycopeptide (LBG), the routinely prepared murine splenic lymphocyte suspension was separately added into the samples with different concentrations (500, 100, 10, 1 μg/ml) of LBG as LBG groups. Blank control group in the absence of Lycium Barbarum glycopeptide or ConA and positive control group in the presence of 0.5 ml ConA but in the absence of LBG were created. 0.5 ml LBG samples with different concentrations in combination with 0.5 ml ConA (10 μg/ml) into each well to observe the synergic effects of LBG and ConA as LBG+ConA groups. After incubation for 72 h at 37 ℃, the samples were analyzed by CFSE-labeled cells combined with flow cytometry, and MTT. Flow cytometry revealed that both LBG could enhance the murine splenic lymphocyte proliferative reaction. Combined use of LBG and ConA had synergic effects. MTT demonstrated that sample A could obviously promote the murine splenic lymphocyte proliferative reaction as compared with control group (P<0.01), while sample B could also enhance the lymphocyte proliferation at a high dose. In combination with ConA, sample A had synergic effects at high dose, while sample B showed obviously synergic effects (P<0.05). It was concluded that both samples (A and B) had strong immunocompetence.
基金supported by Jilin Provincial Science & Technology Department(No.20190201079JC)the Fundamental Research Funds for the Central Universities,JLUOpen Project of State Key Laboratory of Supramolecular Structure and Materials, Jilin University,China(No.sklssm2019020)
文摘Investigations of glycosylated proteins or peptides and their related biological pathways provide new possibilities for illuminating the physiological and pathological mechanisms of glycosylation modification.However,open-ended and in-depth analysis of glycoproteomics is usually subjected to the low-abundance of glycopeptides,heterogeneous glycans,and a variety of interference molecules.In order to alleviate the influence of these obstacles,effective preconcentration of glycopeptides are indispensable.Here,we employed a hydrophilic interaction liquid chromatography(HILIC)-based method to universally capture glycopeptides.Glutathione modified magnetic nanoparticles(Fe304@AuGSH) were synthesized through a simple process and exploited to enrich glycopeptides from complex samples.The prepared materials showed excellent ability to trap glycopeptides from standard glycoproteins digests,low detection limit(10 fmol/p L),and good selectivity(HRP:BSA=1:100).These results indicated that glutathione-based magnetic nanoparticles synthesized in this work had great potential for glycopeptides enrichment.
基金funded by the Research Excellence Consortium(Konsortium Kecemerlangan Penyelidikan)(KKP/2020/UKMUKM/5/1)(JPT(BKPI)1000/016/018/25(21))the Fundamental Research Grant Scheme(FRGS/1/2019/WAB01/UKM/02/1)。
文摘This study was conducted to recover edible bird’s nest(EBN)hydrolysates from different grades of EBN,including the industrial by-products,using enzymatic treatment.The nutrient,physicochemical properties and antioxidant activities of the recovered hydrolysates at different hydrolysis times were evaluated.Results showed that the recovery yield of enzymatic hydrolysis was above 89%for all grades of EBN and the degree of hydrolysis increased over time.Nitrite content(0.321-0.433 mg/L)was below the permissible tolerance level for all samples.Interestingly,the antioxidant activities(DPPH and ABTS scavenging activities and ferric reducing antioxidant powder(FRAP)activity)were significantly higher(P≤0.05)in hydrolysates recovered from EBN by-products(EBNhC and EBNhD)as compared to the high grade EBN hydrolysates(EBNhA and EBNhB).The in-vitro probiotic activity of EBN and its hydrolysates were examined using the probiotic bacterium Lactobacillus plantarum.Evidently,EBN by-products hydrolysate(EBNhD)recorded the highest number of L.plantarum(1.1×1011 CFU/mL),indicating that low grade EBN has the potential as prebiotic material that promotes probiotic activity.This study demonstrated the concept of using EBN by-products hydrolysates for various applications,such as functional ingredients with enhanced bioactivities,to improve its economic value.
文摘Our previous reports have shown that lamininglycopeptides (LN-GPs), the total glycopeptides prepared from laminin (LN), can prevent the experimental lung metastasis and liver metastasis of mouse cancer cells. In order to explore the anti-metastatic mechanism of LNGPs, we studied the effects of LN-GPs on metastasisrelated behaviors of cancer cells in vitro. LN-GPs did not affect cell survival. However, LN-GPs inhibited cell attachment and spreading Of 5180 cells on LN- and Matrigelsubstrate in dose-dependent and time-dependent manners. Moreover, inhibition of cen attachment and spreading on Matrigel substrates were much greater on Matrigel substrate than on LN substrate. In the presence of LN-GPs, 5180 cells on LN substrate changed from a flattened polygonal shape to a round one, the migration of 5180 cells on LN substrate decreased, and the number of a highly invasive human pulmonary giant caxcinoma PG cells invading Matrigel filter in a Boyden chamber was reduced. LN-GPs thus have multiple inhibitory effects on cancer motastasisrelated behaviors.
基金supported by the Natural Science Foundation of Jilin Provincial Science&Technology Department(No.20190201079JC)。
文摘The analysis of endogenous glycoproteins and glycopeptides in human body fluids is of great importance for screening and discovering disease biomarkers with clinical significance.However,the presence of interfering substances makes the direct quantitative detection of low-abundance glycoproteins and glycopeptides in human body fluids one of the great challenges in analytical chemistry.Magnetic solid phase extraction(MSPE)has the advantages of easy preparation,low cost and good magnetic responsiveness.Magnetic adsorbents are the core of MSPE technology,and magnetic adsorbents based on different functional materials are widely used in the quantitative analysis of glycoproteins and glycopeptides in human body fluids,making it possible to analyze glycoproteins and glycopeptides with low abundance as well as multiple types,which provides a technical platform for screening and evaluating glycoproteins and glycopeptides in body fluids as disease biomarkers.In this paper,we focus on the recent advances in the application of MSPE technology and magnetic adsorbents for the separation and enrichment of glycoproteins and glycopeptides in human body fluids,and the future trends and application prospects in this field are also presented.
基金financially supported by Open Project of State Key Laboratory of Supramolecular Structure and Materials,Jilin University,China(No.sklssm2022012)the Fundamental Research Funds for the Central Universities,JLU,China。
文摘Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor selectivity. The present study is the first attempt to combine chitosan(CS) and L-cysteine(L-Cys) to design a novel hydrophilic material focusing on glycopeptide enrichment. CS containing a large number of hydrophilic amino and hydroxyl groups has unique chemical properties, which makes it a very attractive biomaterial for glycopeptide enrichment. The excellent hydrophilicity of zwitterionic molecule L-Cys inspires the idea of anchoring L-Cys onto CS to design a novel hydrophilic material(named as Fe_(3)O_(4)@CS@Au-L-Cys) for the capture of low abundance glycopeptides. To be specific, Au nanoparticles(Au NPs) was introduced into CS-coated Fe_(3)O_(4)via electrostatic interaction and served as bridges to anchor L-Cys onto the surface of CS through strong Au-S bond interaction. The prepared Fe_(3)O_(4)@CS@AuL-Cys exhibited strong affinity, low detection limit(0.5 fmol/μL HRP), high selectivity(HRP/BSA with a molar ratio of 1:1000) for glycopeptides. Moreover, successful application of glycopeptide enrichment in human serum and saliva by Fe_(3)O_(4)@CS@Au-L-Cys was achieved. A satisfactory data set indicates that Fe_(3)O_(4)@CS@Au-L-Cys has promising potential in the application of glycopeptide enrichment in real complex bio-samples and for related glycoproteome research.
基金Supported by the National Science & Technology Pillar Program of China(No2007BA138B00)
文摘A mixture of oligosaccharide and glycopeptide was isolated from the aqueous extract of Panax ginseng roots. The mixture inhibits protein tyrosinc phosphatase（SHP-1） function, implying it enhances immune activity. The peak molecular mass of the oligosaccharide portion is 1800 calculated via GPC software after separation by HPLC. And the structure of the oligosaccharide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→4）- and （1→6）-linked glucopyranoside, with non-reducing terminals of arabinopyranoside and glucopyranoside. The peak molecular mass of glycopeptide portion is 1900 calculated via GPC software after separation by HPLC. The structure of glycopeptide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→3,6）-linked glucopyranoside, with non-reducing terminals of galactopyranose and glucopyranoside. The peptide composition is Glu. Asp, Hyp, Set, Arg, Gly , Thr, Pro, Ala, Val, lie, Leu and Lys. The oligosaccharide-peptide linkage is formed by Ara and Hyp.
基金supported by National Key R&D Program of China (No. 2018YFA0507501)the National Science Foundation for Distinguished Young Scholars of China (No. 21425518)+1 种基金the National Natural Science Foundation of China (Nos. 22074019, 22004017)Shanghai Sailing Program (No. 20YF1405300)。
文摘Exosome and inclusive cargoes have manifested significant function in different biological events. In particular, glycopeptides in exosome are closely associated with occurrence and development of various diseases. Developing advanced tools is highly desired to enrich glycopeptides from exosomes, and enrich exosomes from complex biological samples as well. In this work, integration of L-cysteine and titania onto the surface of magnetic nanoparticles is designed to realize the coefficient affinity towards exosomes and inclusive glycopeptides. Benefiting from the synergistic affinity, we separate exosomes from human urine concentrate directly, which was proved by the detection of three typical antigen markers of exosomes. Furthermore, hardly any exosomes remained on materials after ultrasonication, which confirmed the good capture performance of Fe_(3)O_(4)@TiO_(2)@L-Cys and high release effect of direct lysis.Moreover, 146 glycopeptides corresponding to 77 glycoproteins were successfully identified from captured exosomes. These satisfactory results will inspire more efforts to be devoted to this field and will be extremely helpful to in-depth information excavation of biological markers, especially disease-related ones, through exosomes and inclusive glycopeptides.
基金financially supported by the Fundamental Research Funds for the Central Universities, Jilin University, China。
文摘β-Cyclodextrin(β-CD) based materials have attracted great attention in the separation of hydrophilic glycopeptides due to the abundant hydroxyl groups in its exterior. However, the current materials based on β-CD generally has complex synthesis process and harsh experimental conditions, on the other hand,the interior cavity of β-CD is hydrophobic and is harmful to capture glycopeptides. Herein, a novel hydrophilic material based on β-CD was engineered via a self-assembly process utilizing L-cysteine(L-Cys)or glutathione(GSH) derived adamantane for highly efficient glycopeptide enrichment. It is the first attempt to make use of the hydrophobic interior cavity of β-CD for hydrophilic glycopeptide capture. Taking advantages of strong hydrophilicity and superparamagnetism, the as-prepared materials possess low detection limit, high selectively, and excellent reusability when employed to glycopeptide enrichment. In addition, the feasibility of the hydrophilic material based on β-CD was verified by enriching glycopeptides from human serum and saliva samples. This study provides a heuristic strategy for the application of β-CD-based self-assembly materials in the enrichment of glycopeptides. Importantly, this strategy certified a possible that the change of glycopeptide enrichment sites through host-vip interaction between β-CD and adamantane derivatives with different functional groups.
基金grants from the National Science Foundation of China(81173376).
文摘Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction methods,namely,water extraction,ultrasonic extraction,enzymatic hydrolysis-water extraction,and enzymatic hydrolysis-ultrasonic extraction were considered to determine the best extraction method.Single factor and orthogonal experiments were performed to determine the optimum extracting conditions.Sevage,enzymatic hydrolysis-Sevage,trichloroacetic acid(TCA),TCA-Sevage and enzymatic hydrolysis-TCA methods were tested to determine the best deproteinization method.The glycopeptide content and protein removal ratio were analyzed by the phenol-sulfuric acid and Coomassie Brilliant Blue methods.Results:Gekko sulfated glycopeptide obtained by water extraction could inhibit the proliferation of HepG2 and SKBR3,as well as promote that of lymphocytes.The glycopeptide content was 4.049%in the optimal extracting condition of a triple decoction extraction for 1 hour each time with a material to solvent ratio of 1:15.The enzymatic hydrolysis-TCA method was found to be the optimal deproteinization method,with a protein removal ratio of 50.46%,glycopeptide content of 14.27%,and inhibitory ratio on human HepG2 cells of 49.06%.Conclusion:This extraction and purification technique for Gekko sulfated glycopeptide is reasonable,feasible,and provides a scientific basis for industrial production.
基金supported by National Key R&D Program of China(No.2018YFA0507501)the National Science Foundation for Distinguished Young Scholars of China(No.21425518)+1 种基金the National Natural Science Foundation of China(Nos.22074019,22004017)Shanghai Sailing Program(No.20YF1405300).
文摘Both glycosylation and phosphorylation exert crucial rule in multitudinous biological processes.For in-depth profiling of glycosylation and phosphorylation,a magnetic metal oxide is effectively coupled with inherently hydrophilic mesoporous channels(denoted as Fe_(3)O_(4)@TiO_(2)@mSiO_(2)-TSG).Based on the mechanism of hydrophilic interaction liquid chromatography(HILIC)and metal oxide affinity chromatography(MOAC),the Fe_(3)O_(4)@TiO_(2)@mSiO_(2)-TSG nanomaterial shows high capacity for simultaneously enriching glycopeptides and phosphopeptides.With human saliva collected in successive four days as practical biological sample,endogenous glycopeptides and phosphopeptides are efficiently enriched.Further gene ontology analysis reveals that the identified endogenous glycopeptides and phosphopeptides participate in diverse molecular functions and biological processes.This strategy is anticipated to promote variation analysis of salivary post-translational modifications.
文摘The detailed glycan structural analysis of glycoprotein is amenable to glycopeptide enrichment. Here, we develop a simple, effective and economical approach to enrich glycopeptides from proteolytically digested peptide mixtures by chromatographic column packed with graphite carbon and activated charcoal (G/A-column). Glycopeptide from ovalbumin was efficiently enriched by homemade G/A-column using liquid chromatography and the structure of glycopeptide was obtained by tandem mass spectrometry using Fourier transform ion cyclotron resonance mass spectrometry. The results in this study demonstrate that G/A-column can be used to enrich N-glycolpeptides and be benefit for online identification of glycopeptide using LC-MS.
文摘There are also a variety of cytokines in the tumor microenvironment,which are involved in the change in the hardness of the tumor,thereby affecting the invasion and metastasis of tumor cells.As traditional Chinese medicines,drugs of softening and dissipating firm knot contains different kinds of sulfated polysaccharide and sulfated glycopeptide.By inhibiting the function of cancer-associated fibroblasts,they reduce interstitial fibrosis,thereby reducing the hardness of the tumor and exerting an anti-tumor effect.
基金This work was supported by grants from the New Century Excellent Talent(NCET-11-1068)National Science Foundation of China(No.81173376).
文摘Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting basic fibroblast growth factor(bFGF).bFGF promoted the proliferation of fibroblasts.Both fibrosarcoma and fibroblasts derived from fibrous connective tissue.This study investigated whether GSPP has the inhibitory effects on human fibrosarcoma HT1080 cells.Materials and methods:The trypan blue exclusion assay was used to determine cell viability and cell numbers.Cells migration was observed by wound-healing and transwell.Results:From the first day to seventh day,HT1080 cells number of GSPP,bFGF,GSPP combined bFGF groups had not change compared with control.HT1080 cells migration distance and the number of migrating cells of GSPP,bFGF,GSPP combined bFGF groups were not significantly reduced.Conclusions:GSPP did not have inhibitory effects on the proliferation and migration of human fibrosarcoma HT1080 cells.Thus further research should be carried out in order to study the mechanism of GSPP and bFGF acting on the tumor stroma.
基金This work was supported by grants from the New Century Excellent Talent(NCET-11-1068)and National Science Foundation of China(No.81173376).
文摘Background:Our previous work demonstrated that Gekko sulfated glycopeptide extracted from the Chinese gecko Shou gong(Gekko swinhonis Guenther)could inhibit tumor growth by regulating basic fibroblast growth factor.However,basic fibroblast growth factor has opposing effects on growth in breast and liver cancers.Direct effects and mechanisms of Gekko sulfated glycopeptide on tumor growth remain unclear and are ripe for further exploration.Methods:Differential regulation by Gekko sulfated glycopeptide and bFGF were studied in established human breast cancer MCF-7 cells and hepatocarcinoma HepG2 cells.Cell proliferation was evaluated with a Trypan blue exclusion assay.Cell cycle phases were measured by flow cytometry.Basic fibroblast growth factor,transforming growth factor-β1,and p21WAF1/CIP1 mRNA and protein expression levels were detected by real-time PCR(mRNA)and ELISA(protein).Changes in phosphorylated extracellular signal-regulated kinase levels were analyzed by Western blot.Results:Data indicated that Gekko sulfated glycopeptide inhibited the proliferation of HepG2 cells(P<0.001)and also blocked basic fibroblast growth factor-stimulated proliferation of these cells(P=0.001).Gekko sulfated glycopeptide was shown to increase transforming growth factor-β1 and p21WAF1/CiP1 expression(P<0.01)and partially compensate for reductions therein induced by basic fibroblast growth factor.Conversely,in MCF-7 cells,Gekko sulfated glycopeptide alone had no observable effect on transforming growth factor-β1 and p21WAF1/CiP1 expression.Gekko sulfated glycopeptide did,however,enhance basic fibroblast growth factor-induced inhibition of cell proliferation and transforming growth factor-β1 and p21WAF1/CiP1 expression in the MCF-7 cells(P=0.001,P<0.01,P<0.01,respectively).Gekko sulfated glycopeptide was shown to suppress basic fibroblast growth factor secretion in both HepG2 and MCF-7 cell lines(P<0.05 and P<0.01,respectively)and inhibited extracellular signal-regulated kinase 1/2 phosphorylation facilitated by basic fibroblast growth factor.Gekko sulfated glycopeptide alone decreased phosphorylated extracellular signal-regulated kinase in HepG2 cells but did not visibly affect phosphorylated extracellular signal-regulated kinase levels in MCF-7 cells.Conclusions:Gekko sulfated glycopeptide,a basic fibroblast growth factor inhibitor,differentially regulates growth in different cancer cell lines,and these differences may be determined by the opposing effects of basic fibroblast growth factor on transforming growth factor-β1 and p21WAF1/CiP1 levels in breast and liver cancer cells.
基金supported by grants from National Key R&D Program of China(Grant Nos:2022YFC2502902)National Natural Science Foundation of China(Grant Nos:82072442,82272494,82072082 and 32122046)+1 种基金the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)the Shenzhen Fundamental Research Foundation(Grant Nos.JCYJ20210324115814040 and JCYJ20210324113001005).
文摘Cartilage injury represents a frequent dilemma in clinical practice owing to its inherently limited self-renewal capacity.Biomimetic strategy-based engineered biomaterial,capable of coordinated regulation for cellular and microenvironmental crosstalk,provides an adequate avenue to boost cartilage regeneration.The level of oxidative stress in microenvironments is verified to be vital for tissue regeneration,yet it is often overlooked in engineered biomaterials for cartilage regeneration.Herein,inspired by natural cartilage architecture,a fibril-network glycopeptide hydrogel(Nap-FFGRGD@FU),composed of marine-derived polysaccharide fucoidan(FU)and naphthalenephenylalanine-phenylalanine-glycine-arginine-glycine-aspartic peptide(Nap-FFGRGD),was presented through a simple supramolecular self-assembly approach.The Nap-FFGRGD@FU hydrogels exhibit a native cartilage-like architecture,characterized by interwoven collagen fibers and attached proteoglycans.Beyond structural simulation,fucoidan-exerted robust biological effects and Arg-Gly-Asp(RGD)sequence-provided cell attachment sites realized functional reinforcement,synergistically promoted extracellular matrix(ECM)production and reactive oxygen species(ROS)elimination,thus contributing to chondrocytes-ECM harmony.In vitro co-culture with glycopeptide hydrogels not only facilitated cartilage ECM anabolic metabolism but also scavenged ROS accumulation in chondrocytes.Mechanistically,the chondro-protective effects induced by glycopeptide hydrogels rely on the activation of endogenous antioxidant pathways associated with nuclear factor erythroid 2-related factor 2(NRF2).In vivo implantation of glycopeptide hydrogels successfully improved the de novo cartilage generation by 1.65-fold,concomitant with coordinately restructured subchondral bone structure.Collectively,our ingeniously crafted bionic glycopeptide hydrogels simultaneously rewired chondrocytes’function by augmenting anabolic metabolism and rebuilt ECM microenvironment via preserving redox equilibrium,holding great potential for cartilage tissue engineering.
