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1.
Zhonghua Jie He He Hu Xi Za Zhi ; 42(11): 845-851, 2019 Nov 12.
Artigo em Chinês | MEDLINE | ID: mdl-31694095

RESUMO

Objective: To explore the role of S100A8, the receptor for advanced glycation endproducts (RAGE) and Caveolin-1 in neutrophilic asthmatic rats, and to further study the intervention of roxithromycin and the possible mechanisms. Methods: Male Brown Norway rats were randomly assigned to a control group, an asthma group and a Roxithromycin group. The asthmatic rat model was established by intraperitoneal injection of ovalbumin (OVA) and Freund's complete adjuvant (FCA) mixture, and aerosol inhalation of OVA. Rats in the Roxithromycin group were given roxithromycin injection 30 mg/kg 30 minutes before each challenge. Rats in the control and the asthma groups were replaced with equal volumes of saline, respectively. Bronchoalveolar lavage fluid (BALF) neutrophil percentage (Neu%) and pathological changes of pulmonary tissue (hematoxylin-eosin, HE staining) were measured to confirm the establishment of asthmatic models. The concentration of inflammatory cytokines and S100A8 were quantified by enzyme-linked immunosorbent assay (ELISA), and the expression of Caveolin-1 and RAGE at protein levels were detected by immunohistochemistry and Western blot. Results: Neu% in BALF of the asthma group was significantly higher than those of the control group, and Neu% in the Roxithromycin group was lower than the asthma group (all P<0.01). Pulmonary histology revealed that there were a large number of inflammatory cells infiltrated in the bronchial and perivascular, pulmonary interstitial and alveolar spaces, and the bronchial wall and smooth muscles were thickened obviously in the asthma group. Rats in the Roxithromycin group showed milder inflammation and airway remodeling change than the asthma group. There was no obvious pathological damage in the control group. The concentration of IL-6 and IL-17 in BALF and serum of rats in the asthma group were significantly higher than those in the control group (P<0.01), and Roxithromycin inhibited the high expression of these cytokines (P<0.05). The expression of S100A8 and RAGE in the asthma group were significantly higher than those in the control group [(20.6±4.4) vs (7.1±2.0) ng/L; (885±118) vs (462±102) ng/L; (14.2±1.7) vs (7.6±1.8) ng/L; (774±166) vs (406±69) ng/L, all P<0.05], and Roxithromycin inhibited the high expression of these proteins [(14.3±3.7) vs (20.6±4.4) ng/L; (650±53) vs (885±118) ng/L; (10.4±1.2) vs (14.2±1.7) ng/L; (560±64) vs (728±72) ng/L] (all P<0.05). Meanwhile, the expression of Caveolin-1 in the asthma group was significantly lower than that in the control group (P<0.01), and Roxithromycin up-regulated its expression (P<0.01). Correlation analysis showed that there was a significantly positive correlation between the expression of S100A8 and RAGE (r=0.706, P<0.01), while there was a significantly negative correlation between the expression of S100A8 and Caveolin-1 (r=-0.775, P<0.01), and between the expression of Caveolin-1 and RAGE (r=-0.919, P<0.01). Conclusion: S100A8 and Caveolin-1 may play an important role in neutrophilic asthma via RAGE, and Roxithromycin may exerts anti-inflammatory effects and inhibition of airway remodeling partly through this signaling pathway.


Assuntos
Antibacterianos/farmacologia , Asma/tratamento farmacológico , Calgranulina A/efeitos dos fármacos , Caveolina 1/efeitos dos fármacos , Roxitromicina/farmacologia , Remodelação das Vias Aéreas , Animais , Antibacterianos/administração & dosagem , Western Blotting , Líquido da Lavagem Broncoalveolar , Calgranulina A/metabolismo , Caveolina 1/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Pulmão/fisiopatologia , Masculino , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Ovalbumina , Ratos , Receptor para Produtos Finais de Glicação Avançada , Roxitromicina/administração & dosagem
2.
Sheng Li Xue Bao ; 71(5): 792-798, 2019 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-31646333

RESUMO

Aberrant oxidative metabolism in cells is one of the hallmarks of cancer. Overproduction of reactive species promotes carcinogenesis by inducing genetic mutations and activating oncogenic pathways, and thus, antioxidant therapy is considered as an important strategy for cancer prevention and treatment. Caveolin-1 (Cav-1), a constituent protein of caveolae, is involved in not only the formation of the caveolae, vesicular transport, maintaining cholesterol homeostasis directly, but also many cellular physiological and pathological processes including growth, regulation of mitochondrial antioxidant level, apoptosis and carcinomas by interacting with a lot of signaling molecules through caveolin scaffolding domain. Cav-1 has also been shown to mediate tumor genesis and progression through oxidative stress modulation, while Cav-1-targeted treatment could scavenge the reactive species. Intracellular reactive species could modulate the expression, degradation, post-translational modifications and membrane trafficking of Cav-1. More importantly, emerging evidence has indicated that multiple antioxidants could exert antitumor activities in cancer cells by modulating the signaling of Cav-1. This paper reviewed the research progresses on the roles of Cav-1 and oxidative stress in tumorigenesis and development, and would provide new insights on designing strategies for cancer prevention or treatment.


Assuntos
Caveolina 1 , Neoplasias/patologia , Estresse Oxidativo , Antioxidantes , Apoptose , Carcinogênese , Carcinoma/patologia , Humanos , Mitocôndrias , Transdução de Sinais
3.
Arch Esp Urol ; 72(7): 690-696, 2019 Sep.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-31475680

RESUMO

OBJECTIVE: To compare c-kit-positive interstitial Cajal-like cells (ICC) and Caveolin-1 protein levels as a pacemaker and signaling molecules, on ureteropelvic junction (UPJ) specimens, between two groups of pediatric patients with and without ureteropelvic junction obstruction (UPJO). METHODS: We evaluated the UPJ specimens of 45 pediatric patients operated between 2005- 2012 retrospectively. Group 1 included 37 patients who underwent dismembered pyeloplasty due to UPJO. Eight patients underwent nephrectomy by the other reasons (renal tumor, trauma etc) and had normal UPJ were accepted as Group 2. The specimens were examined immunohistochemically with CD117 and Caveolin-1 antibody. According to the total number of ICC; 0-5 cells were accepted as a few (1), 610 cells as moderate (2), and > 10 as many (3). According to the staining intensity of Caveolin-1 at muscle tissue, a subjective evaluation was performed as; mild staining (1), moderate staining (2) and strong staining (3). RESULTS: The mean value of ICC distribution was calculated 1.37 ± 0.54 in Group 1 and 2.13 ± 0.64 in Group 2 (p = 0.003), and the median value of ICC distribution was found 1 [1-3] in Group 1 and 2 [1-3] in Group 2 (p = 0.008). Median values for the intensity of staining with Caveolin-1 were found 2 [1-3] in the Group 1, and 2.5 [2-3] in the Group 2 (p = 0.025). CONCLUSIONS: A decrease in ICC and Caveolin-1 levels support that there may be a relationship between ICC and Caveolin-1 for UPJO associated with signal transduction and peristalsis in urinary system.


Assuntos
Caveolina 1/metabolismo , Obstrução Ureteral , Criança , Humanos , Pelve Renal , Estudos Retrospectivos , Telócitos , Ureter
4.
DNA Cell Biol ; 38(10): 1048-1055, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31433200

RESUMO

DNA condensed agents can improve the transfection efficiency of the cationic liposome delivery system. However, various condensed agents have distinct transfection efficiency and cellular cytotoxicity. The object of this study was to screen the optimal agents with the high transfection efficiency and low cytotoxicity from four polymer compressive materials, polyethylenimine (PEI), chitosan, poly-l-lysine (PLL), and spermidine. DNA was precompressed with these four agents and then combined to cationic liposomes. Subsequently, the entrapment and transfection efficiency of the obtained complexes were investigated. Finally, the particle sizes, cytotoxicity, and endocytosis fashion of these copolymers (Lipo-PEI, Lipo-chitosan, Lipo-PLL, and Lipo-spermidine) were examined. It was found that these four copolymers had significantly lower cytotoxicity and higher transfection efficiency (45.5%, 42.4%, 36.8%, and 47.4%, respectively) than those in the control groups. The transfection efficiency of Lipo-PEI and Lipo-spermidine copolymers were better than the other two copolymers. In 293T cells, nystatin significantly inhibited the transfection efficiency of Lipo-PEI-DNA and Lipo-spermidine-DNA (51.88% and 46.05%, respectively), which suggest that the endocytosis pathway of Lipo-spermidine and Lipo-PEI copolymers was probably caveolin dependent. Our study indicated that these dual-degradable copolymers especially liposome-spermidine copolymer could be used as the potential biocompatible gene delivery carriers.


Assuntos
Quitosana/química , Lipossomos/química , Polietilenoimina/química , Polilisina/química , Espermidina/química , Transfecção/métodos , Cátions , Caveolina 1/genética , Caveolina 1/metabolismo , Quitosana/metabolismo , Colesterol/química , Colesterol/metabolismo , Endocitose/efeitos dos fármacos , Endocitose/genética , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/metabolismo , Células HEK293 , Humanos , Lipossomos/metabolismo , Nistatina/farmacologia , Tamanho da Partícula , Plasmídeos/química , Plasmídeos/metabolismo , Polietilenoimina/metabolismo , Polilisina/metabolismo , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/metabolismo , Espermidina/metabolismo
5.
Nat Commun ; 10(1): 2692, 2019 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-31217420

RESUMO

Sphingomyelin phosphodiesterase acid-like 3b (SMPDL3b) is a lipid raft enzyme that regulates plasma membrane (PM) fluidity. Here we report that SMPDL3b excess, as observed in podocytes in diabetic kidney disease (DKD), impairs insulin receptor isoform B-dependent pro-survival insulin signaling by interfering with insulin receptor isoforms binding to caveolin-1 in the PM. SMPDL3b excess affects the production of active sphingolipids resulting in decreased ceramide-1-phosphate (C1P) content as observed in human podocytes in vitro and in kidney cortexes of diabetic db/db mice in vivo. Podocyte-specific Smpdl3b deficiency in db/db mice is sufficient to restore kidney cortex C1P content and to protect from DKD. Exogenous administration of C1P restores IR signaling in vitro and prevents established DKD progression in vivo. Taken together, we identify SMPDL3b as a modulator of insulin signaling and demonstrate that supplementation with exogenous C1P may represent a lipid therapeutic strategy to treat diabetic complications such as DKD.


Assuntos
Antígenos CD/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3/metabolismo , Nefropatias Diabéticas/patologia , Insulina/metabolismo , Receptor de Insulina/metabolismo , Esfingomielina Fosfodiesterase/metabolismo , Animais , Caveolina 1/metabolismo , Linhagem Celular , Membrana Celular/metabolismo , Ceramidas/metabolismo , Ceramidas/uso terapêutico , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3/genética , Nefropatias Diabéticas/tratamento farmacológico , Modelos Animais de Doenças , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Podócitos/citologia , Podócitos/metabolismo , Isoformas de Proteínas/metabolismo , Transdução de Sinais , Resultado do Tratamento
6.
Plast Reconstr Surg ; 144(1): 58e-67e, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31246819

RESUMO

BACKGROUND: Fibroproliferative disorders result in excessive scar formation, are associated with high morbidity, and cost billions of dollars every year. Of these, keloid disease presents a particularly challenging clinical problem because the cutaneous scars progress beyond the original site of injury. Altered mechanotransduction has been implicated in keloid development, but the mechanisms governing scar progression into the surrounding tissue remain unknown. The role of mechanotransduction in keloids is further complicated by the differential mechanical properties of keloids and the surrounding skin. METHODS: The authors used human mechanical testing, finite element modeling, and immunohistologic analyses of human specimens to clarify the complex interplay of mechanical stress, strain, and stiffness in keloid scar progression. RESULTS: Changes in human position (i.e., standing, sitting, and supine) are correlated to dynamic changes in local stress/strain distribution, particularly in regions with a predilection for keloids. Keloids are composed of stiff tissue, which displays a fibrotic phenotype with relatively low proliferation. In contrast, the soft skin surrounding keloids is exposed to high mechanical strain that correlates with increased expression of the caveolin-1/rho signaling via rho kinase mechanotransduction pathway and elevated inflammation and proliferation, which may lead to keloid progression. CONCLUSIONS: The authors conclude that changes in human position are strongly correlated with mechanical loading of the predilection sites, which leads to increased mechanical strain in the peripheral tissue surrounding keloids. Furthermore, increased mechanical strain in the peripheral tissue, which is the site of keloid progression, was correlated with aberrant expression of caveolin-1/ROCK signaling pathway. These findings suggest a novel mechanism for keloid progression.


Assuntos
Caveolina 1/fisiologia , Queloide , Mecanotransdução Celular/fisiologia , Transdução de Sinais/fisiologia , Estresse Mecânico , Quinases Associadas a rho/fisiologia , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Análise de Elementos Finitos , Humanos , Queloide/metabolismo , Queloide/fisiopatologia , Masculino , Pessoa de Meia-Idade , Postura/fisiologia , Adulto Jovem
8.
Transplant Proc ; 51(5): 1387-1391, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31036353

RESUMO

AIM: Caveolin-1 (CAV-1) is a molecule associated with endothelial cell dysfunction in chronic antibody-mediated rejection (CAMR) and considered to be a novel biomarker of CAMR. For immunohistochemical staining to reveal CAV-1 expression, most studies have used immunofluorescent stained frozen specimens, whereas formalin-fixed tissues have not been utilized. In the present study, we examined CAV-1 expression in specimens from CAMR patients using an immunoenzymatic technique with formalin-fixed tissues. METHODS: Eleven patients diagnosed with CAMR based on findings of transplanted renal biopsy samples were enrolled. Those biopsy specimens were formalin fixed and stained with CAV-1 using an immunoenzymatic method. Dye extent was evaluated by classifying that in peritubular capillaries (PTC) and glomerular capillaries (GBM) in 3 steps. We then compared the Banff scores for peritubular capillaritis (ptc), glomerulopathy (cg), and C4d using those results. RESULTS: CAV-1 expression was confirmed in vascular endothelium (PTC, GBM), while it was poor in epithelial cells. A Banff score for ptc and cg of 3 points was seen in 3 and 4 cases, of 2 points was seen in 1 and 4 cases, of 1 point was seen in 7 and 3 cases, and of 0 points was seen in 0 and 0 cases, respectively. In PTC, C4d and CAV-1 scores of 3 points were seen in 0 and 9 cases, of 2 points were seen in 2 and 2 cases, of 1 point was seen in 5 and 0 cases, and of 0 points were seen in 4 and 0 cases, respectively. As for GBM, C4d and CAV-1 scores of 3 points were seen in 8 and 7 cases, of 2 points were seen in 2 and 4 cases, of 1 point was seen in 0 and 0 cases, and of 0 points were seen 1 and 0 cases, respectively. CONCLUSION: CAV-1 expression in PTC had a score ≥2 in all cases, indicating that an adequate level of staining of formalin-fixed tissue was attained with the present immunoenzymatic technique. These results suggest that CAV-1 expression examined by the present method may be useful for identifying endothelial dysfunction.


Assuntos
Biomarcadores/análise , Caveolina 1/análise , Rejeição de Enxerto/imunologia , Técnicas Imunoenzimáticas/métodos , Transplante de Rim , Adulto , Biomarcadores/metabolismo , Capilares/metabolismo , Feminino , Fixadores , Formaldeído , Humanos , Isoanticorpos , Glomérulos Renais/patologia , Masculino , Pessoa de Meia-Idade , Fixação de Tecidos/métodos
9.
Int J Oncol ; 54(6): 2054-2068, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31081050

RESUMO

The failure of androgen deprivation therapy in prostate cancer treatment mainly results from drug resistance to androgen receptor antagonists. Although an aberrant caveolin­1 (Cav­1) expression has been reported in multiple tumor cell lines, it is unknown whether it is responsible for the progression of castration­resistant prostate cancer (CRPC). Thus, the aim of the present study was to determine whether Cav­1 can be used as a key molecule for the prevention and treatment of CRPC, and to explore its mechanism of action in CRPC. For this purpose, tissue and serum samples from patients with primary prostate cancer and CRPC were analyzed using immunohistochemistry and enzyme­linked immunosorbent assay, which revealed that Cav­1 was overexpressed in CRPC. Furthermore, Kaplan­Meier survival analysis and univariate Cox proportional hazards regression analysis demonstrated that Cav­1 expression in tumors was an independent risk factor for the occurrence of CRPC and was associated with a shorter recurrence­free survival time in patients with CRPC. Receiver operating characteristic curves suggested that serum Cav­1 could be used as a diagnostic biomarker for CRPC (area under the curve, 0.876) using a cut­off value of 0.68 ng/ml (with a sensitivity of 82.1% and specificity of 80%). In addition, it was determined that Cav­1 induced the invasion and migration of CRPC cells by the activation of the H­Ras/phosphoinositide­specific phospholipase Cε signaling cascade in the cell membrane caveolae. Importantly, simvastatin was able to augment the anticancer effects of androgen receptor antagonists by downregulating the expression of Cav­1. Collectively, the findings of this study provide evidence that Cav­1 is a promising predictive biomarker for CRPC and that lowering cholesterol levels with simvastatin or interfering with the expression of Cav­1 may prove to be a useful strategy with which to prevent and/or treat CRPC.


Assuntos
Antagonistas de Receptores de Andrógenos/farmacologia , Caveolina 1/genética , Caveolina 1/metabolismo , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Neoplasias de Próstata Resistentes à Castração/metabolismo , Sinvastatina/farmacologia , Adulto , Idoso , Caveolina 1/sangue , Linhagem Celular Tumoral , Movimento Celular , Colesterol/sangue , Progressão da Doença , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Estudos Retrospectivos , Análise de Sobrevida , Regulação para Cima/efeitos dos fármacos
10.
Cell Physiol Biochem ; 52(5): 1151-1165, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30990585

RESUMO

BACKGROUND/AIMS: Adipocyte hypertrophy in obesity is associated with inflammation and adipose tissue fibrosis which both contribute to metabolic diseases. Mechanisms regulating lipid droplet expansion are poorly understood. Knock down of the scaffold protein beta 2 syntrophin (SNTB2) increases lipid droplet size of 3T3-L1 adipocytes and the physiological relevance of SNTB2 in adipose tissue morphology and metabolic health was analyzed herein. METHODS: Wild type and SNTB2-/- mice were challenged with 24 weeks high fat diet. Adipose tissue morphology and expression of various genes / proteins including collagens and caveolin-1 was examined. Glucose, insulin, fasting and fed free fatty acids were measured in serum. SNTB2 expression was determined in adipose tissues of patients. RESULTS: Upon high fat diet SNTB2-/- mice displayed reduced adiposity and adipocyte hypertrophy. Expression of various proteins was normal in the different white fat depots of SNTB2-/- mice while caveolin-1 protein and collagen mRNA levels were diminished. Null mice had reduced systemic glucose while fasting and postprandial insulin and insulin response were normal. Fatty acid clearance in the fed state and after insulin injection was enhanced. SNTB2 and caveolin-1 were increased in fat of ob/ob mice. However, no correlation between body mass index and SNTB2 protein in adipose tissues of seven patients was found. In subcutaneous but not in visceral fat the ratio of SNTB2 to alpha syntrophin protein, which affects lipid droplet size in the opposite manner, was associated with BMI. In subcutaneous fat of extremely obese patients SNTB2 mRNA levels were not correlated with weight loss after bariatric surgery. CONCLUSION: Current study shows that high SNTB2 in obese adipose tissues restricts adipocyte growth and thereby may contribute to metabolic diseases.


Assuntos
Adipócitos/metabolismo , Gorduras na Dieta/farmacologia , Proteínas Associadas à Distrofina , Metabolismo dos Lipídeos , Obesidade/metabolismo , Período Pós-Prandial , Adipócitos/patologia , Adulto , Idoso , Animais , Caveolina 1/genética , Caveolina 1/metabolismo , Proteínas Associadas à Distrofina/genética , Proteínas Associadas à Distrofina/metabolismo , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Obesidade/genética , Obesidade/patologia
11.
Biochem Cell Biol ; 97(5): 638-646, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-30986357

RESUMO

We recently demonstrated that Cav1 (caveolin-1) is a negative regulator of Stat3 (signal transducer and activator of transcription-3) activity in mouse fibroblasts and human lung carcinoma SHP77 cells. We now examined whether the cellular context may affect their levels as well as the relationship between them, by assessing Cav1 and Stat3-ptyr705 amounts in different cell lines. In MDA-MB-231, A549, and HaCat cells, Cav1 levels were high and Stat3-ptyr705 levels were low, consistent with the notion of a negative effect of endogenous Cav1 on Stat3-ptyr705 levels in these lines. In addition, manipulation of Cav1 levels revealed a negative effect in MCF7 and mouse fibroblast cells, while Cav1 upregulation induced apoptosis in MCF7 cells. In contrast, however, line MRC9 had high Cav1 and high Stat3-ptyr705 levels, indicating that high Cav1 is insufficient to reduce Stat3-ptyr705 levels in this line. MCF7 and LuCi6 cells had very low Cav1 and Stat3-ptyr705 levels, indicating that the low Stat3-ptyr705 can be independent from Cav1 levels altogether. Our results reveal a further level of complexity in the relationship between Cav1 and Stat3-ptyr705 than previously thought. In addition, we demonstrate that in a feedback loop, Stat3 inhibition upregulates Cav1 in HeLa cells but not in other lines tested.


Assuntos
Neoplasias da Mama/metabolismo , Caveolina 1/metabolismo , Neoplasias Pulmonares/metabolismo , Fator de Transcrição STAT3/metabolismo , Tirosina/metabolismo , Animais , Caveolina 1/antagonistas & inibidores , Células Cultivadas , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C
12.
J Neuroinflammation ; 16(1): 72, 2019 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-30953513

RESUMO

BACKGROUND: Diabetes is known to be a main risk factor of post-stroke hemorrhagic transformation following recombinant tissue plasminogen activator (rt-PA) therapy. However, the mechanism through which diabetes exacerbates hemorrhagic transformation is insufficiently understood. We aimed to verify that CD147, the extracellular matrix metalloproteinase (MMP) inducer, played a vital role in the progress. METHODS: We performed middle cerebral artery occlusion on diabetic and non-diabetic rats, with or without rt-PA treatment, and then compared the glycosylation level of CD147, caveolin-1, MMPs activities, and blood-brain barrier (BBB) permeability. In vitro, tunicamycin treatment and genetic tools were used to produce non-glycosylated and lowly glycosylated CD147. An endogenous glucagon-like peptide-1 receptor (GLP-1R) agonist was used to downregulate the glycosylation of CD147 in vivo. RESULTS: Compared with non-diabetic rats, diabetic rats expressed higher levels of highly glycosylated CD147 in endothelium and astrocytes following rt-PA treatment accompanied by higher activity of MMPs and BBB permeability, in the middle cerebral artery occlusion model. Caveolin-1 was also overexpressed and co-localized with CD147 in astrocytes and endothelium in diabetic rats. In vitro, advanced glycation end products increased the expression of highly glycosylated CD147 in astrocytes and endothelial cells. Downregulating the glycosylation of CD147 lowered the activity of MMPs and promoted the expression of tight junction proteins. The expression of caveolin-1 in endothelial cells and astrocytes was not inhibited by tunicamycin, which revealed that caveolin-1 was an upstream of CD147. In vivo, GLP-1R agonist downregulated the glycosylation of CD147 and further reduced the activity of MMPs and protected the BBB in diabetic rats. CONCLUSION: CD147 is essential for diabetes-associated rt-PA-induced hemorrhagic transformation, and downregulation of CD147 glycosylation is a promising therapy for neurovascular-unit repair after rt-PA treatment of patients with diabetes.


Assuntos
Basigina/metabolismo , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Hemorragia/etiologia , Infarto da Artéria Cerebral Média/complicações , Ativador de Plasminogênio Tecidual/efeitos adversos , Animais , Astrócitos/efeitos dos fármacos , Barreira Hematoencefálica/fisiopatologia , Encéfalo/citologia , Caveolina 1/genética , Caveolina 1/metabolismo , Células Cultivadas , Colágeno Tipo IV/metabolismo , Diabetes Mellitus Experimental/patologia , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Glicosilação/efeitos dos fármacos , Infarto da Artéria Cerebral Média/patologia , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Ocludina/metabolismo , RNA Interferente Pequeno/genética , Ratos , Ratos Wistar , Reperfusão
13.
Cell Commun Signal ; 17(1): 37, 2019 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-30995923

RESUMO

BACKGROUND: We previously showed that caveolin-1 (cav-1), an integral membrane protein, is required for the synthesis of matrix proteins by glomerular mesangial cells (MC). In a previous study to understand how cav-1 is involved in regulating matrix production, we had identified significant upregulation of the antifibrotic protein follistatin in cav-1 knockout MC. Follistatin inhibits the profibrotic effects of several members of the transforming growth factor beta superfamily, in particular the activins. Here, we characterize the molecular mechanism through which cav-1 regulates the expression of follistatin. METHODS: Kidneys from cav-1 wild type and knockout (KO) mice were analyzed and primary cultures of MC from cav-1 wild-type and KO mice were utilized. FST promoter deletion constructs were generated to determine the region of the promoter important for mediating FST upregulation in cav-1 KO MC. siRNA-mediated down-regulation and overexpression of Sp1 in conjunction with luciferase activity assays, immunoprecipitation, western blotting and ChiP was used to assess the role of Sp1 in transcriptionally regulating FST expression. Pharmacologic kinase inhibitors and specific siRNA were used to determine the post-translational mechanism through which cav-1 affects Sp1 activity. RESULTS: Our results establish that follistatin upregulation occurs at the transcript level. We identified Sp1 as the critical transcription factor regulating activation of the FST promoter in cav-1 KO MC through binding to a region within 123 bp of the transcription start site. We further determined that the lack of cav-1 increases Sp1 nuclear levels and transcriptional activity. This occurred through increased phosphoinositide 3-kinase (PI3K) activity and downstream protein kinase C (PKC) zeta-mediated phosphorylation and activation of Sp1. CONCLUSIONS: These findings shed light on the transcriptional mechanism by which cav-1 represses the expression of a major antifibrotic protein, and can inform the development of novel antifibrotic treatment strategies.


Assuntos
Caveolina 1/fisiologia , Folistatina/genética , Regulação da Expressão Gênica , Células Mesangiais/patologia , Fator de Transcrição Sp1/metabolismo , Animais , Caveolina 1/genética , Caveolina 1/metabolismo , Núcleo Celular/metabolismo , Células Cultivadas , Fibrose , Células Mesangiais/metabolismo , Camundongos , Camundongos Knockout , Fosfatidilinositol 3-Quinases/metabolismo , Regiões Promotoras Genéticas , Proteína Quinase C/metabolismo , Transcrição Genética
14.
Gene ; 705: 22-35, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31005612

RESUMO

Mixed-lineage leukaemia 1 (MLL1) enzyme plays major role in regulating genes associated with vertebrate development. Cell physiology and homeostasis is regulated by microRNAs in diverse microenvironment. In this investigation we have identified conserved miR-193a target sites within the 3'-UTR of MLL1 gene transcript. Utilizing wild type and mutated 3'-UTR constructs and luciferase reporter assays we have clearly demonstrated that miR-193a directly targets the 3'-UTR region of the MLL1 mRNA. Ectopic expression of miR-193a modulated global H3K4 mono-, di- and tri-methylation levels and affects the expression of CAV1, a gene which is specifically modulated by H3K4me3. To determine the implications of this in vitro finding in aberrant physiological conditions we analyzed prostate cancer tissue samples. In this context miR-193a RNA was undetectable and MLL1 was highly expressed with concomitantly high levels of H3K4me, H3K4me2, and H3K4me3 enrichment in the promoters of MLL1 responsive genes. Finally, we showed that prolonged ectopic expression of miR-193a inhibits growth and cell migration, and induces apoptosis. Thus, while our study unveils amplitude of the epigenome, including miRnome it establishes that; (i) miR-193a directly target MLL1 mRNA, (ii) miR-193a impair MLL1 protein production, (iii) miR-193a reduces the overall methylation marks of the genome.


Assuntos
Histona-Lisina N-Metiltransferase/genética , Histona-Lisina N-Metiltransferase/metabolismo , Histonas/metabolismo , MicroRNAs/genética , Proteína de Leucina Linfoide-Mieloide/genética , Proteína de Leucina Linfoide-Mieloide/metabolismo , Neoplasias da Próstata/genética , Regiões 3' não Traduzidas , Caveolina 1/genética , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Cromatina/metabolismo , Regulação para Baixo , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Metilação , Neoplasias da Próstata/metabolismo
15.
Am J Chin Med ; 47(3): 675-689, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30966770

RESUMO

Pancreatic cancer cells overexpress the insulin receptor (IR) and the insulin-like growth factor-1 receptor (IGF1R). Activating these receptors, insulin and insulin-like growth factor-1 increase the growth and glycolysis of pancreatic cancer cells. The high glycolysis in pancreatic cancer cells increases whole-body energy expenditure and is therefore involved in the pathogenesis of cancer cachexia. The antagonism of IR and IGF1R may sabotage pancreatic cancer cells and attenuate cancer cachexia. Previous studies have shown that the intracellular regulating system of IR/IGF1R may be functionally interrelated to another intracellular system whose master regulator is hypoxia-inducible factor-1 (HIF-1). In this study, we investigated how the IR/IGF1R and HIF-1 systems are interrelated in pancreatic cancer cells. We also investigated whether a phytochemical, penta-O-galloyl- ß -D-glucose ( ß -PGG), antagonizes IR/IGF1R, sabotages pancreatic cancer cells and alleviates cancer cachexia. We found in MiaPaCa2 pancreatic cancer cells that IR/IGF1R activation increased both the α -subunit of HIF-1 and caveolin-1. This result suggests that IR/IGF1R, HIF-1 α , and caveolin-1 may constitute a feed-forward loop to mediate the effect of IR/IGF1R activation. ß -PGG inhibited IR/IGF1R activity and decreased glycolytic enzymes in MiaPaCa2 and Panc-1 pancreatic cancer cells. When MiaPaCa2 cells were transplanted in athymic mice, their growth was inhibited by ß -PGG or by a HIF-1 α inhibitor, rhein. ß -PGG and rhein also decreased glycolytic enzymes in the tumor grafts and reduced liver gluconeogenesis, skeletal-muscle proteolysis and fat lipolysis in the tumor carriers. Cancer-induced body-weight loss, however, was prevented by ß -PGG but not rhein. In conclusion, ß -PGG combats pancreatic cancer cells and cures cancer cachexia.


Assuntos
Caquexia/tratamento farmacológico , Taninos Hidrolisáveis/farmacologia , Taninos Hidrolisáveis/uso terapêutico , Neoplasias Pancreáticas/metabolismo , Animais , Caquexia/etiologia , Caveolina 1/metabolismo , Glicólise , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Neoplasias Pancreáticas/complicações , Receptor de Insulina/antagonistas & inibidores , Receptor de Insulina/metabolismo , Receptores de Somatomedina/antagonistas & inibidores , Receptores de Somatomedina/metabolismo , Células Tumorais Cultivadas
16.
Toxicol In Vitro ; 57: 132-142, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30825645

RESUMO

A recent epidemiological study suggested that chronic exposure to cleaning detergents significantly reduced lung function in consumers. In this study, we identified the toxic mechanism of ammonium lauryl sulfate (ALS), the most common detergent in consumer products, using alveolar macrophage cells. In preliminary tests, cell viability sharply decreased between 40 and 200 µg/mL, thus we determined doses of 10, 20, and 50 µg/mL for further study. When treated at a 50 µg/mL for 24 h, cell viability was 67.7 ±â€¯3.4% of the control, and autophagosome-like vacuoles and a number of double membranes surrounding damaged mitochondria were observed in the cytosol. Intracellular ROS, the ATP amount, ER volume, acid cell compartments and mitochondrial potential rapidly reduced with dose, whereas the release of LDH and apoptotic bodies dramatically increased. Additionally, multiple cell death pathways were activated following exposure to ALS, and the expression of caveolin-1, p-Acetyl CoA carboxylase, p21, and p-ERK were greatly inhibited. Moreover, the secretion of inflammatory mediators and expression of innate- and adaptive-immune response-related proteins were remarkably reduced. Meanwhile, the secretion of TGF-ß was enhanced. Therefore, we conclude that ALS-induced apoptosis may be due to mitochondrial dysfunction triggered by the inhibition of caveolin-1, and that chronic pulmonary exposure to ALS may cause adverse health effects such as cancer and fibrosis by impairing the host's pulmonary immune system.


Assuntos
Caveolina 1/antagonistas & inibidores , Detergentes/toxicidade , Mitocôndrias/efeitos dos fármacos , Dodecilsulfato de Sódio/toxicidade , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Camundongos , Mitocôndrias/patologia , Espécies Reativas de Oxigênio/metabolismo
17.
Prep Biochem Biotechnol ; 49(5): 453-458, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30896287

RESUMO

Existing evidence has demonstrated liposomes as the gene transporter induce the cytotoxicity during the transfection process through several known pathways. In the present study, we investigated the possibility of siRNAs targeting 3-ß-hydroxysterol △-24-reductase (DHCR24), which encodes an enzyme catalyzing the last step of cholesterol biosynthesis, to suppress the liposome cytotoxicity induced by lipid-based transfection reagent in the neuroblastoma cell line N2A. We found that the siRNAs targeting DHCR24 mRNA protect cells from the liposome-induced cell death, probably through the effect of siDHCR24s on the reduction of the cellular cholesterol and decrease in the generation of reactive oxygen species (ROS). This suggests that siRNAs targeting DHCR24 or other methods that reduce the intracellular cholesterol levels might be a good strategy for avoiding the cytotoxicity of liposomes, without impairing its efficiency of gene-delivering.


Assuntos
Sobrevivência Celular/genética , Colesterol/deficiência , Lipossomos/efeitos adversos , Proteínas do Tecido Nervoso/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-CH/genética , Interferência de RNA , Animais , Caveolina 1/genética , Linhagem Celular Tumoral , Regulação para Baixo , Camundongos , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Transfecção/métodos
18.
Eur J Pharmacol ; 853: 121-128, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30880179

RESUMO

This study examined the mechanism associated with the endothelium-dependent attenuation of vasoconstriction induced by bupivacaine (BPV), with a particular focus on the upstream cellular signaling pathway of endothelial nitric oxide synthase (eNOS) phosphorylation induced by BPV in human umbilical vein endothelial cells (HUVECs). BPV concentration-response curves were investigated in the isolated rat aorta. The effects of Nω-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), methylene blue, calmidazolium, the Src kinase inhibitor 4-amino-3-(4-chlorophenyl)-1-(t-butyl)-1H-pyrazolo[3,4-d]pyrimidine, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) and the combination of L-arginine and L-NAME on BPV-induced contraction in endothelium-intact aorta preparations were examined. The effects of BPV alone and in combination with PP2 on the phosphorylation of eNOS (at Ser1177 or Thr495), caveolin-1 and Src kinase were examined in HUVECs. BPV-induced contraction was lower in endothelium-intact aortae than in endothelium-denuded aortae. L-NAME, ODQ, methylene blue and calmidazolium increased BPV-induced contraction in endothelium-intact aortae, whereas PP2 alone and combined treatment with L-arginine and L-NAME inhibited BPV-induced contraction. Low-concentration BPV (30 µM) induced both stimulatory (Ser1177) and inhibitory (Thr495) phosphorylation of eNOS in HUVECs. However, high-concentration BPV (150 µM) induced only stimulatory (Ser1177) eNOS phosphorylation. Additionally, phosphorylation of Src kinase, caveolin-1 and inhibitory eNOS (Thr495) induced by low-concentration BPV was inhibited by PP2. These results suggest that contraction induced by low-concentration BPV is attenuated by endothelial nitric oxide release, which is modulated both stimulatory (Ser1177) and inhibitory eNOS phosphorylation (Thr495). BPV-induced phosphorylation of eNOS (Thr495) is indirectly mediated by an upstream cellular signaling pathway involving Src kinase (Tyr416) and caveolin-1 (Tyr14).


Assuntos
Bupivacaína/farmacologia , Óxido Nítrico Sintase Tipo III/metabolismo , Vasoconstrição/efeitos dos fármacos , Animais , Aorta/efeitos dos fármacos , Aorta/enzimologia , Aorta/fisiologia , Caveolina 1/metabolismo , Relação Dose-Resposta a Droga , Masculino , Óxido Nítrico Sintase Tipo III/química , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Quinases da Família src/metabolismo
19.
Phytomedicine ; 58: 152888, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30901662

RESUMO

BACKGROUND: A Lung cancer death account for approximately 1 in 5 of all cancer-related deaths and is particularly virulent due to its enhanced metastasis and resistance to chemotherapy. Chrysotobibenzyl has been reported to decrease cell metastasis, according to the results of an anchorage-independent growth assay; however, its underlying mechanism has not been investigated yet. PURPOSE: The aim of this study was to investigate the effect of chrysotobibenzyl on lung cancer cell migration and drug sensitization and its mechanism. METHODS: Cell viability, cell proliferation and drug sensitization were determined by MTT assay. Cell migration was analyzed using a wound-healing assay. Transwell migration and invasion were analyzed using Boyden chamber assay. Mechanisms of chrysotobibenzyl against metastasis including cell migration, invasion, and epithelial to mesenchymal transition (EMT) were evaluated by Western blot analysis and immunofluorescence. RESULTS: Treatment with chrysotobibenzyl was applied at concentrations of 0-50 µM and the results showed non-cytotoxicity in human lung cancer cells (H460, H292, A549, and H23) and other non-cancerous human cells (HCT116, primary DP1 and primary DP2). However, 50 µM of chrysotobibenzyl significantly altered cell proliferation in H292 cells at 48 h. In addition, 1-50 µM of chrysotobibenzyl significantly inhibited H460 and H292 cell migration, invasion, filopodia formation, and decreased EMT in a dose-dependent manner at 48 h, which were correlated with reduced protein levels of integrins ß1, ß3, and αν, p-FAK, p-AKT, Cdc42, and Cav-1. We also established shRNA-Cav-1-transfected (shCav-1) H460 and H292 cells. shCav-1 transfected cells can decrease cell migration and downregulate the expression of integrins ß1, ß3, and αν when compared with the control. Moreover, chrysotobibenzyl was shown to suppress EMT indicated by the reduction of EMT markers (Vimentin, Snail, and Slug), and sensitize lung cancer cells to cisplatin-mediated apoptosis. CONCLUSION: Treatment with chrysotobibenzyl inhibited lung cancer cell migration via Cav-1, integrins ß1, ß3, and αν, and EMT suppressions. The downregulation of integrins in response to the compound not only inhibited cell metastasis, but also sensitized lung cancer cells to cisplatin-mediated apoptosis.


Assuntos
Antineoplásicos/farmacologia , Bibenzilas/farmacologia , Caveolina 1/metabolismo , Integrinas/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/farmacologia , Regulação para Baixo/efeitos dos fármacos , Interações de Medicamentos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/patologia , Pseudópodes/efeitos dos fármacos
20.
Int J Mol Med ; 43(5): 2064-2074, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864740

RESUMO

The aim of the present study was to examine the protective effect of caveolin­1 (Cav­1) in the penehyclidine hydrochloride (PHC)­based inhibition of lipopolysaccharide (LPS)­induced acute lung injury (ALI) in vivo and in vitro, in addition to the potential underlying mechanisms. In vivo, an ALI rat model was established via intratracheal administration of LPS (5 mg/kg), and PHC (2 mg/kg) was administered 30 min following LPS treatment. In vitro, the Cav­1 gene was knocked down by small interfering (si)RNA in J774A.1 cells. Cells were incubated with LPS (1 µg/ml) for 2 h, and subsequently incubated with PHC (2 µg/ml) for an additional 2 h. Lung injury was assessed by lung histology and the ratio of polymorphonuclear leukocytes (PMNs) to total cells was assessed in bronchoalveolar lavage fluid (BALF), myeloperoxidase (MPO) activity, BALF protein content and lung wet/dry (W/D) ratio. The levels of pro­inflammatory factors, including tumor necrosis factor­α (TNF­α), interleukin (IL)­6 and IL­1ß, in the sera of rats and cell culture supernatant were determined by ELISA. The protein expression levels of Cav­1, toll­like receptor 4 (TLR4), phosphorylated (p)­p38 mitogen activated protein kinases (p38 MAPKs) and nuclear factor kappa­light­chain­enhancer of activated B cells transcription factor p65 subunit (NF­κB p65) in lung tissues and J774A.1 cells were analyzed by western blot analysis. The results indicated that PHC effectively alleviated lung injury by decreasing neutrophil infiltration and protein concentration in BALF, and the lung W/D ratio and MPO activity and pro­inflammatory cytokine production induced by LPS. Furthermore, PHC significantly decreased the degrees of histopathological changes and pulmonary dysfunction. In vitro, treatment with PHC inhibited pro­inflammatory cytokine levels and MPO activity in LPS­stimulated J774A.1 cells. However, the results in the J774A.1 cells with Cav­1 gene knockdown were contrary. In addition, PHC decreased TLR4, p­p38 MAPKs and nuclear NF­κB p65 expression levels and upregulated the expression level of Cav­1, in vivo and in vitro. These data demonstrated that PHC exhibited a protective effect against LPS­induced ALI in rats and LPS­stimulated J774A.1 cells, which may be due to the inhibition of p38 MAPKs phosphorylation and TLR4/NF­κB signaling pathway by Cav­1 upregulation.


Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Lesão Pulmonar Aguda/metabolismo , Caveolina 1/metabolismo , Quinuclidinas/uso terapêutico , Regulação para Cima , Animais , Gasometria , Permeabilidade Capilar/efeitos dos fármacos , Linhagem Celular , Citocinas/biossíntese , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pulmão/ultraestrutura , Masculino , Camundongos , Peroxidase/metabolismo , Pneumonia/patologia , Quinuclidinas/farmacologia , Ratos Sprague-Dawley , Regulação para Cima/efeitos dos fármacos
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