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1.
Artigo em Inglês | MEDLINE | ID: mdl-31990059

RESUMO

Aberrant expression of microRNA-876-5p (miR-876-5p) is implicated in the progression of multiple human cancers. However, the potential role of miR-876-5p in colorectal cancer remains poorly understood. The purpose of the current study was to investigate the potential role of miR-876-5p in colorectal cancer. miR-876-5p expression was significantly downregulated in colorectal cancer tissues and cell lines compared with normal controls. Gain-of-function assays revealed that miR-876-5p overexpression effectively repressed the malignant behaviours of colorectal cancer cells, including cell proliferation, colony formation, and invasion. Bioinformatics analysis predicted that RAS protein activator like 2 (RASAL2), a potential oncogene for colorectal cancer, is a putative miR-876-5p target gene. A luciferase reporter assay confirmed that miR-876-5p directly binds to the 3'-untranslated region (UTR) of RASAL2. Furthermore, both RASAL2 messenger RNA (mRNA) and protein expression were negatively modulated by miR-876-5p in colorectal cancer cells. Notably, there was an inverse correlation between miR-876-5p and RASAL2 expression in colorectal cancer tissue specimens. Moreover, miR-876-5p was involved in regulating the activation of Yes-associated protein (YAP) signalling through inhibiting RASAL2. However, the miR-876-5p-mediated antitumour effect on colorectal cancer cells was partially reversed by restoring RASAL2 expression. Notably, miR-876-5p upregulation impeded the tumour growth of colorectal cancer cells in vivo in nude mice. Overall, these results demonstrated that miR-876-5p exerts an antitumour function in colorectal cancer by targeting RASAL2 to suppress YAP signalling activation. These findings highlight the importance of the miR-876-5p/RASAL2/YAP axis in colorectal cancer progression and suggest that miR-876-5p is a potential therapeutic target for treating colorectal cancer.

2.
Biochem Biophys Res Commun ; 521(4): 1017-1023, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31727365

RESUMO

Dysregulation of apoptotic and autophagic function are characterized as the main pathogeneses of diabetic nephropathy (DN). It has been reported that Karyopherin Alpha 2 (KPNA2) contributes to apoptosis and autophagy in various cells, but its role in DN development remains unknown. The purpose of present study was to explore the function and underling mechanisms of KPNA2 in development of DN. In this study, 30 mM high glucose (HG)-evoked podocytes were used as DN model. The expression of KPNA2 was detected by qRT-PCR and Western blot assays. The cell viability was tested by CCK-8 kit, the apoptosis was measured using flow cytometry assay, the apoptotic and the autophagy related genes was detected by Western blot. Our results indicated that KPNA2 was significantly increased after HG stimulation. Knockdown of KPNA2 inhibited apoptosis, and promoted cell viability and autophagy in HG-treated podocytes. In addition, silencing of KPNA2 deactivated mTORC1/p70S6K pathway activation via regulating SLC1A5. Further results demonstrated that activating mTORC1/p70S6K pathway strongly ameliorated the effect of KPNA2 on cell viability, apoptosis and autophagy. Therefore, our study suggested that knockdown of KPNA2 rescued HG-induced injury via blocking activation of mTORC1/p70S6K pathway by mediating SLC1A5.

3.
Lab Invest ; 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31641222

RESUMO

Hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein (HIP/PAP) has antimicrobial, antioxidant, anti-inflammatory, mitogenic, and antiapoptotic effects and thus exerts important functions in the maintenance of integrity and homeostasis of several organs, such as the gastrointestinal tract, pancreas, and liver. Although the potent hepatoprotective effect of HIP/PAP has been validated, its impact on liver fibrosis has not been reported. In this study, we evaluated the role of HIP/PAP on hepatic fibrosis and explored the possible underlying mechanisms. We found that the expression of HIP/PAP and its mouse counterpart, Reg3B, was markedly upregulated in fibrotic human or mouse livers. Intraperitoneal (i.p.) interleukin (IL)-10, IL-6, and TNF-α but not TGF-ß1 significantly induced hepatic overexpression of Reg3B in mice. In both CCl4 and BDL liver fibrosis models, adenovirus-mediated ectopic expression of HIP/PAP markedly alleviated liver injury, inflammation, collagen deposition, hepatic stellate cell activation, and the overexpression of profibrotic cytokines, including transforming growth factor ß1 (TGF-ß1), platelet-derived growth factor (PDGF)-A, B, connective tissue growth factor (CTGF), and plasminogen activator inhibitor-1 (PAI-1), in mice. In vitro experiments demonstrated that, in addition to suppressing hepatic stellate cell proliferation and accelerating hepatocyte proliferation, HIP/PAP mitigated TGF-ß1-induced hepatic stellate cell activation, hepatocyte epithelial-mesenchymal transition (EMT) and upregulated expression of profibrotic cytokines in both hepatic stellate cells and hepatocytes. Moreover, HIP/PAP attenuated the overexpression of TGF-ß receptor II (TGF-ßRII) in fibrotic mouse livers and decreased the basal expression of TGF-ßRII in nonfibrotic mouse livers as well as in cultured hepatocytes and hepatic stellate cells, which is at least partly attributable to the TGF-ß1-antagonizing function of HIP/PAP. This study indicates that increased expression of hepatic HIP/PAP serves as a countermeasure against liver injury and fibrosis. Exogenous supplementation of HIP/PAP might be a promising therapeutic agent for hepatic fibrosis as well as liver injury.

4.
Biochem Biophys Res Commun ; 520(1): 67-72, 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31575408

RESUMO

Intrahepatic cholestasis of pregnancy (ICP) is gestation-specific liver disease associated with liver injury and increased serum and hepatic bile acids. Although the mechanism of ICP is still not fully understood, the reproductive hormones seem to play an important role. Recent studies show that a progesterone metabolite, epiallopregnanolone sulfate (PM5S), is supraphysiologically elevated in the serum of ICP patients, indicating it may play an etiology role in ICP. Bile acid homeostasis is controlled by multiple mechanisms including farnesoid X receptor (FXR)-mediated bile acid export and synthesis. It is known that cholic acid (CA), a primary bile acid, can activate FXR, which is inhibited by PM5S, an FXR antagonist. Here we employed a mouse model of concurrent exposure of CA and PM5S-induced liver injury and determined the effects of probiotic Lactobacillus rhamnosus GG (LGG) in the prevention of the bile acid disorders and liver injury. Mice challenged with CA + PM5S had significantly increased levels of serum and hepatic bile acids and bilirubin and liver enzyme. Pretreatment with LGG significantly reduced bile acid and bilirubin levels associated with reduced liver enzyme level and mRNA expression levels of pro-inflammatory cytokines. We also showed that the beneficial effects of LGG is likely mediated by hepatic FXR activation and bile salt export pump (BSEP) upregulation. In conclusion, our results provide a rationale for the application of probiotics in the management of ICP through gut microbiota-mediated FXR activation.

5.
Theor Appl Genet ; 132(11): 3201-3221, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31501915

RESUMO

KEY MESSAGE: Five putative candidate genes for OWBM resistance in Chinese winter wheat 'Jimai 24' were identified via BSR-seq and differential expression analyses. Orange wheat blossom midge (OWBM), Sitodiplosis mosellana, is one of the most serious threats to wheat production worldwide. Conventional gene mapping methods to identify genes require significant amounts of financial support and time. Here, bulked segregant RNA-seq (BSR-seq) was applied to profile candidate genes and develop associated markers for OWBM resistance. Previously, we identified a major QTL (QSm.hebau-4A) for OWBM resistance on the long arm of chromosome 4A. In this study, we aimed at screening differentially expressed resistance genes associated with this QTL. Twelve differentially expressed genes (DEGs) were obtained based on BSR-seq and differential expression analyses. Among them, four were confirmed to be associated with OWBM resistance via quantitative reverse transcription PCR, using an additional set of wheat samples subjected to OWBM invasion. One SPI-like gene and one Malectin-like gene were revealed by gene annotation, respectively. Sequencing results confirmed that the four DEGs and the SPI gene had SNP polymorphisms between wheat parents. All these five resistance-related genes for OWBM were located in the same genomic region with QSm.hebau-4A. Furthermore, six new markers developed based on sequences of the five genes were also mapped in the same genomic region using genetic population. These five genes may be the candidate genes for OWBM resistance in Chinese wheat 'Jimai 24' and should be the targets for further positional isolation.

6.
J Cell Physiol ; 234(8): 12821-12827, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30741411

RESUMO

BACKGROUND: Osteoporosis (OP) is a systemic osteopathy with increased bone fragility and increased risk of fracture. Osteoclasts (OC) are the key target cells in the treatment of osteoporosis. We aimed to research the role of L-type calcium channel protein Cav1.3 in OC differentiation in this study. METHODS: OP rat model was established to detect the expression level of Cav1.3. Tartrate-resistant acid phosphatase assay was used to measure the differentiation of osteoclast during receptor activator of nuclear factor κ-Β ligand (RANKL)-induced osteoclasts formation. The expression of bone differentiation-related proteins were detected by western blot analysis. RESULTS: Cav1.3 is upregulated in OP rats. Knockdown of Cav1.3 inhibits the differentiation of RAW264.7. Cav1.3 regulates the cell differentiation and bone resorption of RAW264.7 during RANKL-induced osteoclasts formation, which is accompanied by upregulation of CaMK II, p-CERB, AP-1, NFATC1, and NF-κB. CONCLUSION: Cav1.3 plays an important role in osteoporosis and the differentiation of osteoclast, which might be involved with the bone differentiation-related proteins.

7.
BMC Cell Biol ; 19(1): 27, 2018 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-30545286

RESUMO

BACKGROUND: Within the past years, umbilical cord (UC) and amniotic membrane (AM) expanded in human platelet lysate (PL) have been found to become increasingly candidate of mesenchymal stromal cells (MSCs) in preclinical and clinical studies. Different sources of MSCs have different properties, and lead to different therapeutic applications. However, the similarity and differences between the AMMSCs and UCMSCs in PL remain unclear. RESULTS: In this study, we conduct a direct head-to-head comparison with regard to biological characteristics (morphology, immunophenotype, self-renewal capacity, and trilineage differentiation potential) and immunosuppression effects of AMMSCs and UCMSCs expanded in PL. Our results indicated that AMMSCs showed similar morphology, immunophenotype, proliferative capacity and colony efficiency with UCMSCs. Moreover, no significantly differences in osteogenic, chondrogenic and adipogenic differentiation potential were observed between the two types of cells. However, AMMSCs exhibited higher PGE2 expression and IDO activity compared with UCMSCs when primed by IFN-γ and (or) TNF-α induction, and AMMSCs showed a higher inhibitory effect on PBMCs proliferation than UCMSCs. CONCLUSION: The results suggest that AMMSCs expanded in PL showed similar morphology, immunophenotype, self-renewal capacity, and trilineage differentiation potential with UCMSCs. However, AMMSCs possessed superior immunosuppression effects in comparison with UCMSCs. These results suggest that AMMSCs in PL might be more suitable than UCMSCs for treatment of immune diseases. This work provides a novel insight into choosing the appropriate source of MSCs for treatment of immune diseases.


Assuntos
Âmnio/citologia , Células-Tronco Mesenquimais/citologia , Cordão Umbilical/citologia , Adolescente , Adulto , Idoso , Biomarcadores/metabolismo , Plaquetas/metabolismo , Diferenciação Celular , Extratos Celulares , Autorrenovação Celular , Forma Celular , Condrogênese , Humanos , Imunossupressão , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Osteogênese , Adulto Jovem
8.
J Gene Med ; 20(9): e3043, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29972714

RESUMO

BACKGROUND: The present study aimed to clarify the effects of thymosin ß4 (Tß4) on CCl4 -induced hepatic fibrosis in mice and to further explore the underlying mechanisms. METHODS: Expression of Tß4 in fibrotic liver tissues was assessed by a quantitative real time-reverse transcriptase polymerase chain reaction and immunohistochemistry. The effects of intraperitoneal adeno-associated virus-Tß4 (AAV-Tß4) on CCl4 -induced hepatic fibrosis were observed by the evaluation of collagen deposition, hepatic stellate cell (HSC) activation and pro-fibrotic cytokine expression. In vitro tests with HSCs and hepatocytes were performed to confirm the effects of Tß4. RESULTS: The expression of Tß4 was down-regulated in fibrotic mouse livers but was rapidly up-regulated by CCl4 -induced acute injury. AAV-Tß4 pre-treatment significantly attenuated liver injury, collagen deposition, HSC activation and pro-fibrotic cytokine over-expression, such as transforming growth factor ß1 (TGF-ß1), platelet-derived growth factor B (PDGF-B), connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) in CCl4 -intoxicated mouse livers. In vitro experiments showed that Tß4 suppressed HSC proliferation, blunted TGF-ß1-induced HSC activation and reduced TGF-ß1-induced TGF-ß1, PDGF-B, CTGF and PAI-1 expression in both HSCs and hepatocytes. Ectopic Tß4 ameliorated the over-expression of TGF-ß receptor-II (TGF-ßRII) in the fibrotic mouse livers. Exogenous Tß4 down-regulated TGF-ßRII expression, whereas neutralizing endogenous extracellular Tß4 with a specific antibody up-regulated TGF-ßRII expression in cultured HSCs and hepatocytes. CONCLUSIONS: Tß4 possesses anti-fibrotic activity in the liver, which is attributable, at least partly, to down-regulating TGF-ßRII and thereby blunting TGF-ß1-mediated fibrogenetic signaling in both HSCs and hepatocytes.


Assuntos
Regulação da Expressão Gênica/genética , Cirrose Hepática/genética , Receptor do Fator de Crescimento Transformador beta Tipo II/genética , Timosina/genética , Animais , Tetracloreto de Carbono , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Dependovirus/genética , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/metabolismo , Masculino , Camundongos , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo , Timosina/metabolismo , Timosina/farmacologia , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/farmacologia
9.
Clin Rheumatol ; 37(1): 205-212, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29067587

RESUMO

Systemic lupus erythematosus (SLE) is a chronic systemic autoimmune disease, characterized by B cell hyperactivity and pathogenic autoantibodies formation. The objective of this study is to evaluate the distribution of B cell subsets in patients with SLE. We included patients with SLE followed in First Affiliated Hospital of Xi'an JiaoTong University, Xi'an, China. Flow cytometry was used to measure frequencies of B cell subsets, including memory B cells, switched memory B cells, non-switched memory B cells, double-negative memory B cells, and naïve B cells in 130 patients with SLE and 55 healthy controls. The different distributions of B cell subsets were further evaluated by their associations with disease activity and clinical manifestation. SLE patients showed significant alteration of B cell subsets, including lower frequency of non-switched memory B cells and higher double-negative memory B cells. The frequencies of B cell subsets also varied between new-onset SLE patients and chronic SLE patients. Frequencies of total memory B cells, switched memory B cells, and non-switched memory B cells were lower in new-onset SLE patients and frequency of naïve B cells was higher compared with healthy controls. Chronic SLE patients showed increased switched memory B cells and double-negative memory B cells. In addition, switched memory B cells and double-negative B cells were higher in patients with lupus nephritis (LN) regardless of disease activity. Our findings suggested that abnormalities of the B cell subsets homeostasis might contribute to the pathogenesis of SLE.


Assuntos
Subpopulações de Linfócitos B/imunologia , Linfócitos B/imunologia , Memória Imunológica , Lúpus Eritematoso Sistêmico/imunologia , Nefrite Lúpica/imunologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
10.
Org Biomol Chem ; 15(30): 6323-6327, 2017 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-28730195

RESUMO

The Cu(i)-catalyzed stereoconvergent borylative cyclization of ω-mesylate-α,ß-unsaturated compounds is facilitated by a simple Cu-bisphosphine catalyst. This reaction provides a novel route to cis-ß-boron-substituted five- and six-membered carbocycle and heterocycle esters. Mechanistic studies indicate that stereoconvergence and cis-substitution likely stem from the rapid enolation of the borylcopper adduct with the substrate double bond and the formation of a five-membered intermediate, respectively.

11.
Cell Physiol Biochem ; 42(1): 145-155, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28528333

RESUMO

OBJECTIVE: This study aims to investigate the regulatory mechanism of 1,25-(OH)2D3 on the proliferation of fibroblast-like synoviocytes (FLS) and expressions of pro-inflammatory cytokines in rheumatoid arthritis (RA) rats via microRNA-22 (miR-22). METHODS: A rat model of RA was established with a subcutaneous injection of type II collagen. After treated with different concentrations of 1,25-(OH)2D3 the proliferation of FLS was estimated by the MTT method, and the optimal concentration of 1,25-(OH)2D3 was selected for further experiments. Cell proliferation was detected by MTT. Cell cycle and apoptosis were analyzed by FCM. The IL-1ß, IL-6, IL-8, and PGE2 protein expressions were determined by ELISA, and MMP-3, INOS, and Cox-2 mRNA expressions were measured by qRT-PCR. RESULTS: The rat model of RA was successfully established. Compared with the blank group, the 1,25-(OH)2D3 and miR-22 inhibitors groups exhibited higher proliferation inhibition and apoptosis rates, lower levels of pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, and PGE2), and decreased mRNA expressions of MMP-3, INOS, and Cox-2. The miR-22 mimics group had lower proliferation inhibition and apoptosis rates, elevated expressions of pro-inflammatory cytokines and MMP-3, INOS, and Cox-2 than the blank group. In contrast to the 1,25-(OH)2D3 group, the proliferation inhibition and apoptosis rates were down-regulated, and the expressions of pro-inflammatory cytokines and MMP-3, INOS, and Cox-2 were up-regulated in the 1,25-(OH)2D3 + miR-22 mimics group. CONCLUSION: Our study demonstrated that 1,25-(OH)2D3 inhibits the proliferation of FLS and alleviates inflammatory response in RA rats by down-regulating miR-22.


Assuntos
Artrite Reumatoide/patologia , Colecalciferol/farmacologia , Citocinas/análise , MicroRNAs/metabolismo , Animais , Antagomirs/metabolismo , Apoptose , Artrite Reumatoide/genética , Artrite Reumatoide/metabolismo , Proliferação de Células , Células Cultivadas , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Dinoprostona/metabolismo , Modelos Animais de Doenças , Regulação para Baixo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Masculino , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Sprague-Dawley , Sinoviócitos/citologia , Sinoviócitos/patologia
12.
Molecules ; 22(5)2017 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-28452949

RESUMO

Gremlin-1, a highly conserved glycosylated and phosphorylated secretory protein, plays important roles in diverse biological processes including early embryonic development, fibrosis, tumorigenesis, and renal pathophysiology. Aptamers, which are RNA or DNA single-stranded oligonucleotides capable of binding specifically to different targets ranging from small organics to whole cells, have potential applications in targeted imaging, diagnosis and therapy. In this study, we obtained a DNA aptamer against Gremlin-1 (G-ap49) using in vitro Systematic Evolution of Ligands by Exponential Enrichment (SELEX). Binding assay and dot-blot showed that G-ap49 had high affinity for Gremlin-1. Further experiments indicated that G-ap49 was quite stable in a cell culture system and could be used in South-Western blot analysis, enzyme-linked aptamer sorbent assay (ELASA), and aptamer-based cytochemistry and histochemistry staining to detect Gremlin-1. Moreover, our study demonstrated that G-ap49 is capable of revealing the subcellular localization of Gremlin-1. These data indicate that G-ap49 can be used as an alternative to antibodies in detecting Gremlin-1.


Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Animais , Aptâmeros de Nucleotídeos/síntese química , Sequência de Bases , Células HEK293 , Humanos , Immunoblotting , Camundongos , Microscopia de Fluorescência , Técnica de Seleção de Aptâmeros , Coloração e Rotulagem
13.
World J Gastroenterol ; 23(2): 242-255, 2017 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-28127198

RESUMO

AIM: To investigate the protective effect of a recombinant adeno-associated virus carrying thymosin ß4 (AAV-Tß4) on murine colitis via intracolonic administration. METHODS: AAV-Tß4 was prepared and intracolonically used to mediate the secretory expression of Tß4 in mouse colons. Dextran sulfate sodium (DSS) was applied to induce the murine ulcerative colitis, and 2,4,6-trinitrobenzene sulfonic acid (TNBS) was used to establish a mouse colitis model resembling Crohn's disease. The disease severity and colon injuries were observed and graded to reveal the effects of AAV-Tß4 on colitis. The activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were determined using biochemical assays. Colonic levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-10 were measured using ELISA, and mucosal epithelial cell apoptosis and proliferation were detected by TUNEL assay and immunochemistry, respectively. RESULTS: Recombinant AAVs efficiently delivered LacZ and Tß4 into the colonic tissues of the mice, and AAV-Tß4 led to a strong expression of Tß4 in mouse colons. In both the DSS and TNBS colitis models, AAV-Tß4-treated mice displayed distinctly attenuated colon injuries and reduced apoptosis rate of colonic mucosal epithelia. AAV-Tß4 significantly reduced inflammatory cell infiltrations and relieved oxidative stress in the inflamed colons of the mice, as evidenced by decreases in MPO activity and MDA content and increases in SOD activity. AAV-Tß4 also modulated colonic TNF-α, IL-1ß and IL-10 levels and suppressed the compensatory proliferation of colonic epithelial cells in DSS- and TNBS-treated mice. CONCLUSION: Tß4 exerts a protective effect on murine colitis, indicating that AAV-Tß4 could potentially be developed into a promising agent for the therapy of inflammatory bowel diseases.


Assuntos
Colite Ulcerativa/metabolismo , Colo/metabolismo , Doença de Crohn/metabolismo , Mucosa Intestinal/metabolismo , Timosina/metabolismo , Animais , Proliferação de Células , Colite Ulcerativa/induzido quimicamente , Colo/enzimologia , Doença de Crohn/induzido quimicamente , DNA Recombinante/administração & dosagem , Dependovirus/genética , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Enterócitos/metabolismo , Enterócitos/fisiologia , Ensaio de Imunoadsorção Enzimática , Vetores Genéticos/administração & dosagem , Imunoquímica , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/enzimologia , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Peroxidase/metabolismo , Superóxido Dismutase/metabolismo , Ácido Trinitrobenzenossulfônico/toxicidade , Fator de Necrose Tumoral alfa/metabolismo
14.
Sci Rep ; 6: 32155, 2016 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-27562139

RESUMO

Connective tissue growth factor (CTGF) has been recognized as a central mediator and promising therapeutic target in hepatic fibrosis. In this study, we generated a novel virus-like particle (VLP) CTGF vaccine by inserting the 138-159 amino acid (aa) fragment of CTGF into the central c/e1 epitope of C-terminus truncated hepatitis B virus core antigen (HBc, aa 1-149) using a prokaryotic expression system. Immunization of BALB/c mice with the VLP vaccine efficiently elicited the production of anti-CTGF neutralizing antibodies. Vaccination with this CTGF vaccine significantly protected BALB/c mice from carbon tetrachloride (CCl4)-induced hepatic fibrosis, as indicated by decreased hepatic hydroxyproline content and lower fibrotic score. CCl4 intoxication-induced hepatic stellate cell activation was inhibited by the vaccination, as indicated by decreased α-smooth muscle actin expression and Smad2 phosphorylation. Vaccination against CTGF also attenuated the over-expression of some profibrogenic factors, such as CTGF, transforming growth factor-ß1, platelet-derived growth factor-B and tissue inhibitor of metalloproteinase-1 in the fibrotic mouse livers, decreased hepatocyte apoptosis and accelerated hepatocyte proliferation in the fibrotic mouse livers. Our results clearly indicate that vaccination against CTGF inhibits fibrogenesis, alleviates hepatocyte apoptosis and facilitate hepatic regeneration. We suggest that the vaccine should be developed into an effective therapeutic measure for hepatic fibrosis.


Assuntos
Intoxicação por Tetracloreto de Carbono/prevenção & controle , Fator de Crescimento do Tecido Conjuntivo/farmacologia , Antígenos do Núcleo do Vírus da Hepatite B/farmacologia , Cirrose Hepática/prevenção & controle , Vacinas/farmacologia , Vírion , Animais , Intoxicação por Tetracloreto de Carbono/metabolismo , Intoxicação por Tetracloreto de Carbono/patologia , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/imunologia , Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/imunologia , Cirrose Hepática/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/farmacologia , Vacinas/imunologia
15.
J Gene Med ; 18(10): 261-272, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27572454

RESUMO

BACKGROUND: Extracellular high mobility group box 1 (HMGB1) is crucially implicated in the pathogenesis of inflammatory bowel diseases (IBDs). A box domain of HMGB1 has been identified as a specific antagonist of HMGB1. In the present study, we tested the effects of adeno-associated virus (AAV)-mediated colonic secretory expression of HMGB1 A box on murine experimental colitis. METHODS: Self-complementary AAV-2 carrying mouse immunoglobin Gκ leader-human HMGB1 A box (AAV-HMGB1 A box) was constructed. The effects of intracolonically administered AAV-HMGB1 A box on dextran sulfate sodium (DSS)- and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis were assessed by the disease activity index (DAI), colon length, macroscopic and histological scoring, myeloperoxidase (MPO) activity, and epithelial apoptosis and complementary proliferation. Colonic immune cell infiltrates, mucosal malondialdehyde content and superoxide dismutase activity, colonic tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-10 levels, serum HMGB1 concentration, and colonic HMGB1 release were determined to investigate the underlying mechanisms. RESULTS: Intracolonically administered AAV-HMGB1 A box efficiently mediated secretory expression of HMGB1 A box and led to significant decreases in DAI, macroscopic and histological scores and colonic epithelial apoptosis in both DSS- and TNBS-treated mice. Modulating inflammation-associated cytokines, such as inhibiting colonic TNF-α and IL-1ß expression, decreasing HMGB1 release, and restoring colonic IL-10 levels, and thereby inhibiting inflammatory cell infiltration and alleviating oxidant damage, might be the underlying mechanism. CONCLUSIONS: Intracolonic application of AAV-HMGB1 A box is effective in alleviating murine colitis and has therapeutic potential in human IBDs.


Assuntos
Colite/genética , Colo/metabolismo , Dependovirus/genética , Proteína HMGB1/genética , Animais , Apoptose/genética , Proliferação de Células/genética , Colite/induzido quimicamente , Colite/metabolismo , Citocinas/metabolismo , Sulfato de Dextrana , Células Epiteliais/metabolismo , Proteína HMGB1/metabolismo , Humanos , Masculino , Camundongos Endogâmicos BALB C , Peroxidase/metabolismo , Superóxido Dismutase/metabolismo , Ácido Trinitrobenzenossulfônico
16.
Artif Cells Nanomed Biotechnol ; 44(5): 1333-8, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27328726

RESUMO

Exogenous insulin and EGFP genes were transduced into bone marrow mesenchymal stem cells of beagle dogs using the retroviral vector pMSCV to prepare ß-like cells. These cells were autotransplanted into the liver of diabetic dogs through hepatic arterial intervention, and physiological indices before and after transplantation were monitored. Autotransplantation of ß-like cells significantly improved blood sugar, insulin levels, and body mass of diabetic dogs (P < 0.01) continuously for over 80 weeks. Since the liver function remained normal and no tumors formed, this method was determined to be reliable and safe. Intrahepatic autotransplantation of ß-like cells had long-term, reliable, and safe therapeutic effects on diabetic dogs.


Assuntos
Diabetes Mellitus Experimental/terapia , Artéria Hepática , Células Secretoras de Insulina/transplante , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Animais , Autoenxertos , Cães
17.
Exp Ther Med ; 11(2): 645-649, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26893660

RESUMO

The aim of the present study was to investigate the association between serum vascular endothelial growth factor (VEGF) and insulin-like growth factor 1 (IGF-1) levels and the prognosis of patients with uterine fibroids following uterine artery embolization (UAE) treatment. A total of 70 patients with uterine fibroids and 20 healthy controls were enrolled in this study between 2012 and 2014. The serum levels of IGF-1 and VEGF were measured using ELISA. Multiple-factor analysis was performed to assess the association between serum levels of IGF-1/VEGF and certain clinical characteristics, including size, location, number of uterine fibroids and adenomyosis. Progression-free survival curves were analyzed using the Kaplan-Meier method. The serum levels of IGF-1 and VEGF in patients with uterine fibroids prior to UAE treatment were significantly higher than those in controls (P<0.05). At 1 week after UAE treatment, the serum levels of IGF-1 and VEGF were significantly lower compared with those prior to UAE treatment. The serum levels of IGF-1 and VEGF at 1 or 3 months after UAE treatment were significantly higher than those at 1 week after UAE treatment. The serum levels of IGF-1 and VEGF were significantly correlated with the clinical characteristics of uterine fibroids (P<0.05). Lower levels of IGF-1 and VEGF in the serum following UAE treatment were associated with an enhanced progression-free survival of patients. In conclusion, the levels of IGF-1 and VEGF in the serum following UAE treatment can be used as indicators of prognosis in patients with uterine fibroids.

18.
Int J Clin Exp Med ; 8(6): 9549-54, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26309622

RESUMO

AIMS: This study is to investigate the expression levels of vascular endothelial growth factor (VEGF) and cancer antigen 125 (CA125) in serum of adenomyosis patients before and after interventional therapy. The role of serum levels of VEGF and CA125 for the prognosis of adenomyosis is further studied. METHODS: A total of 80 adenomyosis patients treated with interventional therapy and 40 healthy individuals were enrolled in this study. Enzyme-linked immunosorbent assay was performed to detect the expression levels of VEGF and CA125. Receiver operating characteristic analysis was used to determine the treatment effect on adenomyosis. Kaplan-Meier analysis was used to analysis the progression-free survival curve for prognosis of adenomyosis. RESULTS: The expression levels of VEGF and CA125 in serum of patients with adenomyosis was increased when compared with those of healthy individuals before interventional therapy (P < 0.05). Levels of hemoglobin in adenomyosis patients after surgery was increased compared with those before surgery (P < 0.05). The blood volume of menstruation, pain intensity, and volume of uterus in adenomyosis patients after surgery was significantly decreased when compared with those before surgery (P < 0.01). The survival rate of adenomyosis patients with high VEGF and CA125 levels was decreased. Serum levels of VEGF and CA125 had a high sensitivity and specificity for the prognosis of adenomyosis. CONCLUSIONS: The serum expression levels of VEGF and CA125 are related to the development of adenomyosis. VEGF and CA125 serum levels can be used for predicting the prognosis of adenomyosis.

19.
Biomed Res Int ; 2015: 510514, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25738154

RESUMO

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by chronic inflammation. Different studies have shown decreased bone mineral density (BMD) in patients with SLE. The objective of this study was to investigate the prevalence and possible risk factors of low BMD in untreated female patients with SLE in Chinese population. A total of 119 untreated female patients with SLE were included. BMD was measured at lumbar spine and at total hip by dual-energy X-ray absorptiometry. The associations between decreased BMD and demographic variables, clinical variables, and bone metabolism variables were analyzed. These SLE patients had the following characteristics: mean age was 32.6 ± 11.9 years, mean disease duration was 22.1 ± 34.5 months, and mean SLEDAI was 11.4 ± 5.4. Osteopenia was present in 31.1% of the patients and osteoporosis in 8.5%. A significant negative association between low density lipoprotein cholesterol (LDL-c) and BMD at the lumbar spine (correlation coefficient = -0.242; P = 0.023) and total hip (correlation coefficient = -0.259; P = 0.019) was shown. These results seem to indicate that increased LDL-c may be an important risk factor for low BMD at lumbar spine and total hip in untreated female SLE patients.


Assuntos
Densidade Óssea , Doenças Ósseas Metabólicas/diagnóstico por imagem , Quadril/diagnóstico por imagem , Vértebras Lombares/diagnóstico por imagem , Lúpus Eritematoso Sistêmico/diagnóstico por imagem , Absorciometria de Fóton , Adolescente , Adulto , Idoso , Doenças Ósseas Metabólicas/epidemiologia , Doenças Ósseas Metabólicas/etiologia , Criança , Feminino , Humanos , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/epidemiologia , Pessoa de Meia-Idade , Prevalência , Fatores de Risco
20.
Biochem Biophys Res Commun ; 457(4): 640-6, 2015 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-25603056

RESUMO

Connective tissue growth factor (CTGF) is a secreted matricellular protein possessing complex biological functions. CTGF modulates a number of signaling pathways that are involved in cell adhesion, migration, angiogenesis, myofibroblast activation, extracellular matrix deposition and tissue remodeling. Aptamers are oligonucleic acid chains or polypeptides that bind with specific target molecules hence have the potential to be used in the detection and blockade of the targets. In this study, we selected CTGF-targeting DNA aptamers by using systematic evolution of ligands by exponential enrichment (SELEX). After 8 iterative rounds of selection, cloning, DNA sequencing and affinity determination, six aptamers with high affinities to CTGF were obtained. Among them, one (C-ap17P) binds with the N-terminal region (aa 1-190) and the other five (C-ap11, 12, 14, 15 and 18) bind with the C-terminal region (aa 191-350) of hCTGF specifically. The biological stability assay indicated that a representative aptamer, C-ap17P, could keep its integrity at a rather high level for at least 24 h in complete DMEM cell culture medium. These CTGF aptamers might be used as a easy and fast detection tool for CTGF and be developed as CTGF-specific inhibitors for both research works and clinical applications.


Assuntos
Aptâmeros de Nucleotídeos/química , Fator de Crescimento do Tecido Conjuntivo/análise , Sequência de Bases , Sítios de Ligação , Técnicas Biossensoriais , Humanos , Técnica de Seleção de Aptâmeros/métodos
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