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METRNL is a recently identified secreted protein with emerging functions. This study is to find major cellular source of circulating METRNL and to determine METRNL novel function. Here, we show METRNL is abundant in human and mouse vascular endothelium and released by endothelial cells using endoplasmic reticulum-Golgi apparatus pathway. By creating endothelial cell-specific Metrnl knockout mice, combined with bone marrow transplantation to produce bone marrow-specific deletion of Metrnl, we demonstrate that most of circulating METRNL (approximately 75%) originates from the endothelial cells. Both endothelial and circulating METRNL decrease in atherosclerosis mice and patients. By generating endothelial cell-specific Metrnl knockout in apolipoprotein E-deficient mice, combined with bone marrow-specific deletion of Metrnl in apolipoprotein E-deficient mice, we further demonstrate that endothelial METRNL deficiency accelerates atherosclerosis. Mechanically, endothelial METRNL deficiency causes vascular endothelial dysfunction including vasodilation impairment via reducing eNOS phosphorylation at Ser1177 and inflammation activation via enhancing NFκB pathway, which promotes the susceptibility of atherosclerosis. Exogenous METRNL rescues METRNL deficiency induced endothelial dysfunction. These findings reveal that METRNL is a new endothelial substance not only determining the circulating METRNL level but also regulating endothelial function for vascular health and disease. METRNL is a therapeutic target against endothelial dysfunction and atherosclerosis.
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Objective To explore the expression of CRELD2 at the gene and protein levels of mouse tissues, and to provide a reference for studying the biological function of CRELD2 in various tissues. Methods The expression level of CRELD2 in the liver, pancreas, stomach, and lung of C57BL/6J mice was determined by real-time PCR and Western Blot. Results RT-PCR and WB showed that CRELD2 was expressed in mouse liver, pancreas, stomach, and lung. The relative expression levels of CRELD2 from high to low were pancreas, stomach, liver, and lung at the gene level, and pancreas, liver, stomach, and lung at protein level respectively. The result suggested that the relative expression levels of the CRELD2 gene and protein in different tissues were not completely consistent, suggesting that it is related to transcriptional regulation. Conclusion CRELD2 is expressed in mouse liver, pancreas, stomach, and lung, and the relative expression levels of CRELD2 are not completely parallel at the gene and protein level.
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Gram-positive bacteria secrete virulence factors into host cells and cause suppurative inflammation, which leads to the emergence of diseases, therefore poses a great threat to human health. Identifying secreted proteins is beneficial to understand the secretion system and pathogenic mechanism of bacteria, and lays the foundation for further screening of pathogenic factors. Due to the lack of classical signal peptide sequence in non-classical secreted proteins, it is relatively difficult and time-consuming to identify such proteins in large-scale experiments. At present, some computational prediction methods have been proposed, but their performance in predicting non-classical secreted proteins of Gram-positive bacteria is not satisfactory. This paper proposed an ensemble learning model - SPNG+, which integrates six machine learning algorithms including naive bayes, random forest, support vector machine, two gradient promotion trees XGBoost and LightGBM, and K-nearest neighbor through stacking strategy. The results of 5-fold cross validation and independent dataset test show that the SPNG+ is superior to the single machine learning model, the simple integrated learning model and the existing prediction tools in predicting non-classical secreted proteins of Gram-positive bacteria. Compared with the predictors constructed by limited feature coding methods or single machine learning algorithms in the past, the proposed method is a useful supplement to the study of non-classical secreted proteins in Gram-positive bacteria. The source code of SPNG+ is available from https: / / github.com / weidai00 / SPNG.
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Context: Nanoparticle albumin-bound paclitaxel (Nab-PTX) is a form of paclitaxel bound to albumin nanoparticles and is used widely in a neoadjuvant setting for patients with breast cancer. Aims: We conducted a retrospective study to compare the efficacy and safety of Nab-PTX to PTX as neoadjuvant chemotherapy for patients with operable HER2-negative breast cancer. Settings and Design: In total, 50 patients were enrolled. Nab-PTX was administered in the study group, and PTX was administered in the control group. Subjects and Methods: The clinical response and safety profile were recorded. The expression of secreted protein acidic rich in cysteine (SPARC) in tumor tissue was examined. Statistical Analysis: The efficacy and safety analyses were computed using SPSS statistical software. Multiple logistic regression analysis was performed to evaluate the exploratory variables (age, stage, estrogen receptor, partial response, and SPARC expression) for the pathological complete response (pCR), and Fisher's exact test was performed to evaluate the relationship between SPARC and pCR. Results: Both groups of patients achieved a good clinical response. The pCR rate for the Nab-PTX regimen was significantly higher than that for the PTX regimen. The most common adverse events were neutropenia, peripheral sensory neuropathy, arthralgia, and myalgia. In 68% of cases in the Nab-PTX group, high SPARC expression was observed. Conclusions: As neoadjuvant therapy, the Nab-PTX regimen has advantages over conventional taxane regimen in patients with HER2-negative breast cancer. With this regimen, a high pCR rate was achieved with a good safety profile
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Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein highly expressed in bone tissue that acts as a chemoattractant factor promoting the arrival of prostate cancer (PCa) cells to the bone marrow. However, the contribution of SPARC during the early stages of tumor progression remains unclear. In this study, we show that SPARC is highly expressed in PCa tissues with a higher Gleason score. Through stable knockdown and overexpression of SPARC in PC3 and LNCaP cells, respectively, here we demonstrate that endogenous SPARC induces the epithelial-mesenchymal transition (EMT), decreasing E-cadherin and cytokeratin 18 and increasing N-cadherin and vimentin. Moreover, SPARC induces the expression of EMT regulatory transcription factors Snail family transcriptional repressor 1 (Snail), Snail family transcriptional repressor 2 (Slug), and zinc finger E-box binding homeobox 1 (Zeb1). In addition, SPARC knockdown in PC3 cells decreases migration and invasion in vitro, without modifying cell proliferation. Our results indicate that SPARC might facilitate tumor progression by modifying the cellular phenotype in cancer cells.
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Humans , Male , Blotting, Western , Cell Line, Tumor , Epithelial-Mesenchymal Transition , Neoplasm Grading , Neoplasm Invasiveness , Osteonectin/metabolism , Prostatic Neoplasms/pathology , Tissue Array AnalysisABSTRACT
Objective: To investigate the effects and mechanisms of secreted protein, acidic and rich in cysteine (SPARC) on high fluoride-induced apoptosis of thyrocytes. Methods: Human thyroid cells (Nthy-ori 3-1) were cultured and treated with various concentrations (0.1, 1 and 10 mmol/L) of NaF for 24 h, and the expression of SPARC was evaluated using Real-time PCR and Western blot, respectively. The cells were divided into four groups: control group, NaF group, si-SPARC group (cells were transfected with SPARC siRNA for 48 h and then exposed to NaF for 24 h), and si-NC group (cells were transfected with negative control siRNA for 48 h and then exposed to NaF for 24 h). Cytotoxicity was assayed using CCK-8 and LDH; cell apoptosis rate was detected with ELISA. The expressions of cleaved caspase3 (c-caspase3) and IGF-1R were measured by Western blot. In addition, si-SPARC and si-IGF-1R were co-transfected into thyrocytes to further explore mechanisms of SAPRC by evaluating apoptosis. Results: The mRNA and protein levels of SPARC were augmented with the increase of NaF (P<0.05). Cell viability was significantly higher in si-NC group than that in si-SPARC group [(84.02±9.51)% vs. (58.31±6.86)%, P<0.05], and the release rate of LDH was lower [(134.25±18.98)% vs. (195.18±23.50)%, P<0.05]. Cell apoptosis rate was lower in si-SPARC group than that in si-NC group [(124.67±19.44)% vs. (175.24±16.46)%, P<0.05]. In addition, silencing SPARC upregulated the expression of IGF-1R (1.95±0.24 vs. 0.93±0.08, P<0.05), and inhibition of IGF-1R reversed the effect of SPARC on apoptosis. Conclusion: Inhibition of SPARC reduces high fluoride-induced cytotoxicity and blocks cell apoptosis. The possible mechanism is through the negative regulation of IGF-1R.
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Filtering bleb scarring is the main cause of glaucoma filtration surgery failure. Subconjunctival injection of antimetabolites, such as mitomycin and 5-fluorouracil, is widely used clinically to reduce the incidence of scarring, which improves the success rate of the surgery. However, accompanied side effects such as cytotoxicity should not be ignored. Secreted protein acidic and rich in cysteine (SPARC) as a matricellular protein is widely distributed in the eyes, which plays an important role in the process of wound repairing and tissue remodeling. The expression of SPARC is significantly elevated in the mouse model of subconjunctival scarring. Researches suggest that SPARC participates in and regulate the formation of bleb scarring through multiple pathways, therefore it may become a specific new target in the anti-scarring therapy.
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Objective:To investigate the effects of vascular endothelial growth factor (VEGF) on the expression of secreted protein acidic and rich in cysteine (SPARC) and the fibrosis in cultured human Tenon's fibroblast (HTF) in vitro. Methods:HTF cells were obtained from Tenon's capsule tissues of patients undergoing strabismus surgery. Immunofluorescence was used to identify the HTF cells. HTF cells were cultured with different concentrations of VEGF, and which were divided into four groups, i.e., 0 ng/mL group, 25 ng/mL group, 50 ng/mL group and 100 ng/mL group. The expression of SPARC, collagen- , and matrix metalloprotein 9 (MMP-9) and the activity of extracellular signal-regulated kinase (ERK) pathway were analyzed by Western blotting and real-time quantitative PCR (qPCR). The abilities of proliferation and migration of HTF cells were detected by MTS assay and scratch test, respectively. Results:HTF cells were observed and identified by inverted phase contrast microscope and immunofluorescence. Under the stimulation of VEGF, the expression of protein and mRNA of SPARC, collagen-I and MMP-9 of HTF cells in other three groups were increased compared with 0 ng/mL group; the phosphorylation activities of ERK pathway were up-regulated, and the proliferation and migration abilities of HTF cells were up-regulated. And the effect was the most obvious in the 50 ng/mL group. Conclusion:VEGF is involved in promoting the fibrosis of HTF cells accompanied by the up-regulation of the SPARC, which suggests SPARC may become a potential regulatory site.
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Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein highly expressed in bone tissue that acts as a chemoattractant factor promoting the arrival of prostate cancer (PCa) cells to the bone marrow. However, the contribution of SPARC during the early stages of tumor progression remains unclear. In this study, we show that SPARC is highly expressed in PCa tissues with a higher Gleason score. Through stable knockdown and overexpression of SPARC in PC3 and LNCaP cells, respectively, here we demonstrate that endogenous SPARC induces the epithelial-mesenchymal transition (EMT), decreasing E-cadherin and cytokeratin 18 and increasing N-cadherin and vimentin. Moreover, SPARC induces the expression of EMT regulatory transcription factors Snail family transcriptional repressor 1 (Snail), Snail family transcriptional repressor 2 (Slug), and zinc finger E-box binding homeobox 1 (Zeb1). In addition, SPARC knockdown in PC3 cells decreases migration and invasion in vitro, without modifying cell proliferation. Our results indicate that SPARC might facilitate tumor progression by modifying the cellular phenotype in cancer cells.
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Objective To investigate the secreted protein acidic and rich in cysteine (SPARC) expression in the smooth muscle tissue of ob/ob mice.Methods Study time:February 2017 to June 2017.Six ob/ob mice and 6 littermates aged 10 weeks were selected for the trial.The mesenteric artery smooth muscle tissues were collected.qRT -PCR was used to detect SPARC mRNA expression in smooth muscle tissue.The protein expression of SPARC was assayed by Western blot.Results The SPARC mRNA [(1.73 ± 0.65)] and protein [(1.73 ± 0.65)] in smooth muscle tissue of ob/ob mice presented higher expression compared with those in the littermates [(1.00 ± 0.31),(1.00 ± 0.33),t =8.437,5.533,all P ＜ 0.05].Conclusion The higher expression of SPARC in smooth muscles tissue of ob/ob mice may be involved in diabetes combined with atherosclerosis.
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Objective@#Secreted modular calcium-binding proteins (SMOCs) are extracellular glycoproteins of the secreted protein, acidic, and rich in cysteine-related modular calcium-binding protein family and include two isoforms, SMOC1 and SMOC2, in humans. Functionally, SMOCs bind to calcium for various cell functions. In this review, we provided a summary of the most recent advancements and findings of SMOC1 and SMOC2 in development, homeostasis, and disease states.@*Data sources@#All publications in the PubMed database were searched and retrieved (up to July 24, 2019) using various combinations of keywords searching, including SMOC1, SMOC2, and diseases.@*Study selection@#All original studies and review articles of SMOCs in human diseases and embryo development written in English were retrieved and included.@*Results@#SMOC1 and SMOC2 regulate embryonic development, cell homeostasis, and disease pathophysiology. They play an important role in the regulation of cell cycle progression, cell attachment to the extracellular matrix, tissue fibrosis, calcification, angiogenesis, birth defects, and cancer development.@*Conclusions@#SMOC1 and SMOC2 are critical regulators of many cell biological processes and potential therapeutic targets for the control of human cancers and birth defects.
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Objective To investigate the relationship between the expression of Wnt induced secreted protein-1 (WISP-1) and the fibrosis of renal biopsy tissue in IgA nephropathy (IgAN) patients.Methods Fifty-three patients firstly diagnosed as IgA nephropathy by renal biopsy were included and classified according to Oxford and Lee's classification.Sixteen patients with MCD entered the fibrosis negative control group,and fourteen healthy adults entered the normal control group.The expression of WISP-1 in renal tissues and serum of all subjects were detected by immunohistochemistry and ELISA respectively.Results Immunohistochemistry results showed that WISP-1 was not expressed in MCD patients and normal human kidney tissues,which was abundantly deposited in renal tissue of patients with focal proliferative IgAN with renal interstitial fibrosis.The serum level of WISP-1 in IgAN patients was significantly higher than that in normal subjects (P=0.015) and MCD patients (P=0.030).In the subgroup analysis of IgAN renal fibrosis,the serum concentration of WISP-1 of fibrosis grade between 0-10％ (F1 group) and fibrosis ＞ 25％ (F3 group) were significantly higher than that in the normal group and the MCD group (all P ＜ 0.05).There was no significant difference between F2 group (10％ ＜ fibrosis≤25％) and normal group or MCD group (P ＞ 0.05).Conclusions The expression of WISP-1 in serum and renal tissue of renal interstitial fibrosis IgAN patients is higher than that of normal and MCD patients without renal fibrosis,and the IgAN patients' serum level of WISP-1 is significantly increased in fibrosis lower score group.The expressions of WISP-1 in serum and renal tissue are related to the occurrence of IgAN renal interstitial fibrosis,in which WISP-1 may play an important role as an early precursor factor in the pathogenesis of IgAN renal interstitial fibrosis.
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Objective To investigate the relationship between the expression of Wnt induced secreted protein-1 (WISP-1) and the fibrosis of renal biopsy tissue in IgA nephropathy (IgAN) patients.Methods Fifty-three patients firstly diagnosed as IgA nephropathy by renal biopsy were included and classified according to Oxford and Lee's classification.Sixteen patients with MCD entered the fibrosis negative control group,and fourteen healthy adults entered the normal control group.The expression of WISP-1 in renal tissues and serum of all subjects were detected by immunohistochemistry and ELISA respectively.Results Immunohistochemistry results showed that WISP-1 was not expressed in MCD patients and normal human kidney tissues,which was abundantly deposited in renal tissue of patients with focal proliferative IgAN with renal interstitial fibrosis.The serum level of WISP-1 in IgAN patients was significantly higher than that in normal subjects (P=0.015) and MCD patients (P=0.030).In the subgroup analysis of IgAN renal fibrosis,the serum concentration of WISP-1 of fibrosis grade between 0-10％ (F1 group) and fibrosis ＞ 25％ (F3 group) were significantly higher than that in the normal group and the MCD group (all P ＜ 0.05).There was no significant difference between F2 group (10％ ＜ fibrosis≤25％) and normal group or MCD group (P ＞ 0.05).Conclusions The expression of WISP-1 in serum and renal tissue of renal interstitial fibrosis IgAN patients is higher than that of normal and MCD patients without renal fibrosis,and the IgAN patients' serum level of WISP-1 is significantly increased in fibrosis lower score group.The expressions of WISP-1 in serum and renal tissue are related to the occurrence of IgAN renal interstitial fibrosis,in which WISP-1 may play an important role as an early precursor factor in the pathogenesis of IgAN renal interstitial fibrosis.
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Objective To investigate the clinical values of CA199 ,CA242 and DKK1 in diagnosing pancreatic cancer (PCA). Methods The serum levels of CA199 ,CA242 and DKK1 were measured with ECLIA and ELISA method in 112 patients with PCA ,58 patients with benign pancreatic disease and 62 individuals undergoing physical examination.The Kruskal-Wallis single fac-tor analysis of variance (ANOVA) and Mann-Whitney rank-sum test were adopted to conduct the statistical comparison.The ROC curve was drawn by using the Logistic regression model and the area under curve (AUCROC ) was calculated.Results The levels of serum CA199 ,CA242 and DKK1 in the patients with pancreatic cancer were significantly higher than those in the benign pancreatic disease group and healthy normal control group ,the difference was statistically significant ( H=76.30 ,61.2 ,47.60 ,P<0.05);the sensitivities of CA199 ,CA242 and DKK1 for diagnosing pancreatic cancer were 76.7% ,69.6% and 85.7% respectively ;the speci-ficities were 95.0% ,96.7% and 92.5% respectively ;the accuracies were 86.2% ,83.6% and 89.2% respectively.The sensitivity , specificity and accuracy of combined detection of CA 199 ,CA242 and DKK1 by the Logistic regression equation P= 1/[1 +e-(-4.163+0.21X1+0.156X2+0.342X3) ] were 94.4% ,90.8% and 92.2% respectively ;AUC of CA199 ,CA242 ,DKK1 and pre-1 were 0.736 , 0.862 ,0.886 and 0.949 ,respectively.Conclusion Serum CA199 ,CA242 and DKK1 levels have an important value in the diagnosis of pancreatic cancer ,and their combined detection could significantly improve the sensitivity and accuracy of pancreatic cancer diag-nosis.
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Abnormal expression of secreted protein acidic and rich in cysteine like 1 (SPARCL1 )gene is closely related to the development,metastasis and prognosis of a variety of tumors.Recent studies show that SPARCL1 gene is high expressed in glioma.The protein product of SPARCL1 gene encoded can interact with collagen,thus affecting the metastasis ability of tumor,which is positively correlated with tumor malignant de-gree.SPARCL1 gene is low expressed in gastric cancer,colorectal cancer,prostate cancer and breast cancer, which is negatively correlated with the invasion and metastasis abilities of tumors.At present,the role of speci-fic molecular mechanisms of SPARCL1 gene remains controversial. The expressions and functions of SPARCL1 gene in tumor tissues seem to depend on the tumor microenvironment.
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Secreted protein, acidic, cysteine-rich (SPARC)-related modular calcium binding 1 (SMOC1) has been implicated in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs). In this study, we found that a peptide (16 amino acids in length), which is located in the extracellular calcium (EC) binding domain of SMOC1, stimulated osteogenic differentiation of human BMSCs in vitro and calvarial bone regeneration in vivo. Treatment of BMSCs with SMOC1-EC peptide significantly stimulated their mineralization in a dose-dependent manner without changing their rate of proliferation. The expression of osteogenic differentiation marker genes, including type 1 collagen and osteocalcin, also increased in a dose-dependent manner. To examine the effect of the SMOC1-EC peptide on bone formation in vivo, the peptide was covalently immobilized onto hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) particles. X-ray photoelectron spectroscopy analysis showed that the peptide was successfully immobilized onto the surface of HA/β-TCP. Implantation of the SMOC1-EC peptide-immobilized HA/β-TCP particles into mouse calvarial defects and subsequent analyses using microcomputed tomography and histology showed significant bone regeneration compared with that of calvarial defects implanted with unmodified HA/β-TCP particles. Collectively, our data suggest that a peptide derived from the EC domain of SMOC1 induces osteogenic differentiation of human BMSCs in vitro and efficiently enhances bone regeneration in vivo.
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Animals , Humans , Mice , Amino Acids , Bone Marrow , Bone Regeneration , Calcium , Ceramics , Collagen Type I , In Vitro Techniques , Mesenchymal Stem Cells , Miners , Osteocalcin , Osteogenesis , Photoelectron Spectroscopy , Regeneration , X-Ray MicrotomographyABSTRACT
As the first most common cause of death in China,stroke has become a public health problem that seriously affects national economy and people′s livelihood. Unfortunately,only 3% to 5% of stroke patients receive tissue plasminogen activator(tPA)treatment,the only pharmacological therapy ap?proved for ischemic stroke,and no drug is available for hemorrhagic stroke. Therefore,there is an ur?gent need to develop new drugs for stroke therapy. Despite the awareness that neuroprotective agents could be a common strategy for the treatment of both ischemic and hemorrhagic stroke,numerous neu?roprotective agents have showed failure in clinical trials. Combined with the current therapeutic strategies and drug development of stroke,this paper elaborated the stroke injury mechanisms and corresponding clinical drug research targeting excitotoxicity,oxidative and nitrosative stress,and inflammation. From a new perspective,this paper has proposed a novel therapeutic strategy targeting inherent defense mechanisms against stroke,with nicotinamide phosphoribosyltransferase(Nampt)- nicotinamide ade?nine dinucleotide defense system as an example to present our experimental evidence that Nampt can serve as an anti-stroke target and nicotinamide mononucleotide as an anti-stroke agent under development. It is hoped that the bottleneck of stroke therapy can be overcome with unremitting efforts so as to reduce the financial burden and mental stress,and bring benefits to people around the world.
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Objective:To express secreted protein-pgp3 of Chlamydia trachomatis(Ct)plasmid,produce monoclonal antibodies (mAbs)and identify their basic biological characteristics.Methods: Construction pGEX-6p2-pgp3 prokaryotic expression vector,then expressed GST-pgp3 fusion protein in E.coli as antigen used to immune BALB/c mice, spleen cells were fused with SP2/0 mouse myeloma cells.The hybridoma cell lines of screening mAbs were secreted by ELISA,and mAbs specificity,type,class and titer were identified.Results:GST-pgp3 fusion protein was successful expressed,5 strains stable hybridoma cell lines that secreted mAbs were screened out,including 4 strains secreted anti-pgp3 mAbs(P1B3,P2A1,P2B6,P2C2),mAbs type were IgG1/κ,the other strain secretion anti-GST mAbs(P3B4),mAbs type was IgG2b/κ.The titer of P1B3,P2A1,P2B6,P2C2,P3B4 were 1∶6 400,1∶3 200,1∶12 800,1∶6 400 and 1∶6 400 respectively.Conclusion:Successful prepared anti-pgp3 mAbs,and lay a foundation for further study the function of Ct plasmid protein pgp3 and the establishment of Ct detection method.
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Obesity and obesity-related diseases have become the main threat to human health .Acid secreted proteins that are rich in cysteine mainly derived from fat tissue , and are associated with insulin resistance , diabetes and diabetic nephropathy .This re-view summaries molecular biology features such as resistance of cell adhesion , regulating cell proliferation , tissue differentiation and embryonic development and the latest research progress of its role in the obesity and obesity related diseases .
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Objective To explore the effect of aconitine on serum zinc finger protein 41 and secreted protein acidic and rich in cysteine in ventricular septal defect model rats.Methods 80 ventricular septal defect model rats were randomly divided into two groups, experimental group ( n=40) were treated with 0.05 mg/kg aconitine via tail vein injection for 7 consecutive days;control group (n=40) were treated with normal saline via tail vein injection for 7 consecutive days.Then, the abdominal aorta blood of each rat was collected, and the contents of zinc finger protein 41 and SPARC in two groups were detected by Western blot and ELISA method,seperately.Results Western blot results showed that the expression of zinc finger protein 41 and SPARC in serum samples of experimental group were significantly lower than that of control group ( P<0.05 ) .ELISA results showed that the contents of zinc finger protein 41 and SPARC of experimental group was significantly lower than that of control group ( P<0.05 ), respectively.Conclusion Aconitine can decrease the expression of serum zinc finger protein 41 and SPARC in ventricular septal defect model rat.