Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 26
Filtrar
1.
J Proteome Res ; 23(7): 2532-2541, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38902972

RESUMO

Metabolic dysfunction is recognized as a contributing factor in the pathogenesis of wet age-related macular degeneration (wAMD). However, the specific metabolism-related proteins implicated in wAMD remain elusive. In this study, we assessed the expression profiles of 92 metabolism-related proteins in aqueous humor (AH) samples obtained from 44 wAMD patients and 44 cataract control patients. Our findings revealed significant alterations in the expression of 60 metabolism-related proteins between the two groups. Notably, ANGPTL7 and METRNL displayed promising diagnostic potential for wAMD, as evidenced by area under the curve values of 0.88 and 0.85, respectively. Subsequent validation studies confirmed the upregulation of ANGPTL7 and METRNL in the AH of wAMD patients and in choroidal neovascularization (CNV) models. Functional assays revealed that increased ANGPTL7 and METRNL played a pro-angiogenic role in endothelial biology by promoting endothelial cell proliferation, migration, tube formation, and spouting in vitro. Moreover, in vivo studies revealed the pro-angiogenic effects of ANGPTL7 and METRNL in CNV formation. In conclusion, our findings highlight the association between elevated ANGPTL7 and METRNL levels and wAMD, suggesting their potential as novel predictive and diagnostic biomarkers for this condition. These results underscore the significance of ANGPTL7 and METRNL in the context of wAMD pathogenesis and offer new avenues for future research and therapeutic interventions.


Assuntos
Proteína 7 Semelhante a Angiopoietina , Proteínas Semelhantes a Angiopoietina , Humor Aquoso , Biomarcadores , Degeneração Macular Exsudativa , Humor Aquoso/metabolismo , Humanos , Biomarcadores/metabolismo , Masculino , Degeneração Macular Exsudativa/metabolismo , Degeneração Macular Exsudativa/genética , Feminino , Proteínas Semelhantes a Angiopoietina/metabolismo , Proteínas Semelhantes a Angiopoietina/genética , Neovascularização de Coroide/metabolismo , Neovascularização de Coroide/genética , Neovascularização de Coroide/patologia , Idoso , Proliferação de Células , Animais , Movimento Celular , Camundongos
2.
J Neuroinflammation ; 21(1): 123, 2024 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-38725082

RESUMO

BACKGROUND: Hepatic encephalopathy (HE) is closely associated with inflammatory responses. However, as a crucial regulator of the immune and inflammatory responses, the role of leucine-rich repeat kinase 2 (LRRK2) in the pathogenesis of HE remains unraveled. Herein, we investigated this issue in thioacetamide (TAA)-induced HE following acute liver failure (ALF). METHODS: TAA-induced HE mouse models of LRRK2 wild type (WT), LRRK2 G2019S mutation (Lrrk2G2019S) and LRRK2 knockout (Lrrk2-/-) were established. A battery of neurobehavioral experiments was conducted. The biochemical indexes and pro-inflammatory cytokines were detected. The prefrontal cortex (PFC), striatum (STR), hippocampus (HIP), and liver were examined by pathology and electron microscopy. The changes of autophagy-lysosomal pathway and activity of critical Rab GTPases were analyzed. RESULTS: The Lrrk2-/--HE model reported a significantly lower survival rate than the other two models (24% vs. 48%, respectively, p < 0.05), with no difference found between the WT-HE and Lrrk2G2019S-HE groups. Compared with the other groups, after the TAA injection, the Lrrk2-/- group displayed a significant increase in ammonium and pro-inflammatory cytokines, aggravated hepatic inflammation/necrosis, decreased autophagy, and abnormal phosphorylation of lysosomal Rab10. All three models reported microglial activation, neuronal loss, disordered vesicle transmission, and damaged myelin structure. The Lrrk2-/--HE mice presented no severer neuronal injury than the other genotypes. CONCLUSIONS: LRRK2 deficiency may exacerbate TAA-induced ALF and HE in mice, in which inflammatory response is evident in the brain and aggravated in the liver. These novel findings indicate a need of sufficient clinical awareness of the adverse effects of LRRK2 inhibitors on the liver.


Assuntos
Encefalopatia Hepática , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina , Falência Hepática Aguda , Camundongos Knockout , Tioacetamida , Animais , Camundongos , Encefalopatia Hepática/patologia , Encefalopatia Hepática/genética , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/genética , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/metabolismo , Falência Hepática Aguda/induzido quimicamente , Falência Hepática Aguda/patologia , Falência Hepática Aguda/genética , Camundongos Endogâmicos C57BL , Tioacetamida/toxicidade
3.
Mol Ther ; 30(3): 1252-1264, 2022 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-34999209

RESUMO

Endothelial tip cell specialization plays an essential role in angiogenesis, which is tightly regulated by the complicated gene regulatory network. Circular RNA (circRNA) is a type of covalently closed non-coding RNA that regulates gene expression in eukaryotes. Here, we report that the levels of circMET expression are significantly upregulated in the retinas of mice with oxygen-induced retinopathy, choroidal neovascularization, and diabetic retinopathy. circMET silencing significantly reduces pathological angiogenesis and inhibits tip cell specialization in vivo. circMET silencing also decreases endothelial migration and sprouting in vitro. Mechanistically, circMET regulates endothelial sprouting and pathological angiogenesis by acting as a scaffold to enhance the interaction between IGF2BP2 and NRARP/ESM1. Clinically, circMET is significantly upregulated in the clinical samples of the patients of diabetic retinopathy. circMET silencing could reduce diabetic vitreous-induced endothelial sprouting and retinal angiogenesis in vivo. Collectively, these data identify a circRNA-mediated mechanism that coordinates tip cell specialization and pathological angiogenesis. circMET silencing is an exploitable therapeutic approach for the treatment of neovascular diseases.


Assuntos
Neovascularização de Coroide , Retinopatia Diabética , Animais , Neovascularização de Coroide/genética , Retinopatia Diabética/tratamento farmacológico , Retinopatia Diabética/genética , Células Endoteliais/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , RNA Circular/genética , Proteínas de Ligação a RNA/metabolismo , Retina/metabolismo
4.
Genomics ; 113(3): 1482-1490, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33771636

RESUMO

Retinal ischemia-reperfusion (I/R) is involved in the pathogenesis of many vision-threatening diseases. circRNAs act as key players in gene regulation and human diseases. However, the global circRNA expression profile in retinal I/R injury has not been fully uncovered. Herein, we established a murine model of retinal I/R injury and performed circRNA microarrays to identify I/R-related circRNAs. 1265 differentially expressed circRNAs were identified between I/R retinas and normal retinas. Notably, the detection of cWDR37 level in aqueous humor could discriminate glaucoma patients from cataract patients (AUC = 0.9367). cWdr37 silencing protected against hypoxic stress- or oxidative stress-induced retinal ganglion cell (RGC) injury. cWdr37 silencing alleviated IR-induced retinal neurodegeneration as shown by increased NeuN staining, reduced retinal reactive gliosis, and decreased retinal apoptosis. Collectively, this study provides a novel insight into the pathogenesis of retinal I/R injury. cWdr37 is a promising target for the diagnosis or treatment of I/R-related ocular diseases.


Assuntos
Glaucoma , Traumatismo por Reperfusão , Animais , Apoptose , Glaucoma/genética , Humanos , Camundongos , RNA Circular/genética , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Retina
5.
Mol Ther ; 28(10): 2191-2202, 2020 10 07.
Artigo em Inglês | MEDLINE | ID: mdl-32755566

RESUMO

Epigenetic alterations occur in many physiological and pathological processes. N6-methyladenosine (m6A) modification is the most prevalent modification in eukaryotic mRNAs. However, the role of m6A modification in pathological angiogenesis remains elusive. In this study, we showed that the level of m6A modification was significantly upregulated in endothelial cells and mouse retinas following hypoxic stress, which was caused by increased METTL3 levels. METTL3 silencing or METTL3 overexpression altered endothelial cell viability, proliferation, migration, and tube formation in vitro. METTL3 knockout in vivo decreased avascular area and pathological neovascular tufts in an oxygen-induced retinopathy model and inhibited alkali burn-induced corneal neovascularization. Mechanistically, METTL3 exerted its angiogenic role by regulating Wnt signaling through the m6A modification of target genes (e.g., LRP6 and dishevelled 1 [DVL1]). METTL3 enhanced the translation of LRP6 and DVL1 in an YTH m6A RNA-binding protein 1 (YTHDF1)-dependent manner. Collectively, this study suggests that METTL3-mediated m6A modification is an important hypoxic stress-response mechanism. The targeting of m6A through its writer enzyme METTL3 is a promising strategy for the treatment of angiogenic diseases.


Assuntos
Adenosina/análogos & derivados , Epigênese Genética , Regulação da Expressão Gênica , Metiltransferases/metabolismo , Neovascularização Patológica/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Adenosina/metabolismo , Animais , Biomarcadores , Suscetibilidade a Doenças , Inativação Gênica , Humanos , Hipóxia/complicações , Hipóxia/metabolismo , Camundongos , Camundongos Knockout , Neovascularização Patológica/metabolismo , Doenças Retinianas/etiologia , Doenças Retinianas/metabolismo , Doenças Retinianas/patologia , Via de Sinalização Wnt
6.
Ophthalmic Res ; 63(5): 483-490, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31945770

RESUMO

PURPOSE: Osthole is an agent isolated from Cnidium monnieri (L.) Cusson and has been used to treat several disorders. Corneal neovascularization is a sight-threatening condition associated with several inflammatory or infectious ocular disorders. In this study, we investigated the anti-angiogenic effects of osthole on corneal neovascularization and the underlying mechanism. METHODS: MTT assay, HE staining, and calcein-AM/propidium iodide staining was conducted to detect the toxicity of osthole in vitro and in vivo. Corneal neovascularization of ICR mice was induced by alkali burn and observed by a slit lamp microscopy on day 7 after alkali injury. EdU assay, Ki67 immunofluorescence assay, Transwell migration assay, and Matrigel assay were conducted to investigate the role of osthole in endothelial angiogenic effects in vitro. Western blots were conducted to investigate the anti-angiogenic mechanism of osthole in corneal neovascularization. RESULTS: Administration of osthole ranging from 0.05 to 25 µM had no detectable cytotoxicity or tissue toxicity in vivo and in vitro. Topical administration of osthole inhibited corneal neovascularization induced by alkali burn. Osthole decreased the proliferation, migration, and tube-formation of endothelial cells induced by VEGF. Osthole inhibited endothelial angiogenic functions through blocking the phosphorylation of ERK1/2, JNK, and p38. CONCLUSION: Our study provides evidence that osthole is a promising drug for the treatment of corneal neovascularization.


Assuntos
Córnea/patologia , Neovascularização da Córnea/tratamento farmacológico , Cumarínicos/uso terapêutico , Medicina Tradicional Chinesa/métodos , Adjuvantes Imunológicos/uso terapêutico , Angelica , Animais , Células Cultivadas , Córnea/efeitos dos fármacos , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR
7.
Mol Ther Nucleic Acids ; 30: 407-420, 2022 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-36381574

RESUMO

Transfer RNA-derived fragments (tRFs) are a novel class of non-coding RNA transcripts and play important roles in several physiological/pathological processes. However, the role of tRFs in ocular angiogenesis remains elusive. Herein, we investigate whether the intervention of tRF-1001 expression could suppress pathological ocular angiogenesis. The results show that the levels of tRF-1001 expression were reduced in the retinas of an oxygen-induced retinopathy (OIR) model, choroidal neovascularization model, and endothelial sprouting model in vitro. Increased tRF-1001 expression could suppress ocular angiogenesis and endothelial sprouting in vivo and reduce endothelial migration, specification, and sprouting in vitro. Mechanistically, tRF-1001 regulated endothelial angiogenic effects via tRF-1001/METTL3/RBPJ-MAML1 signaling. The levels of tRF-1001 expression were downregulated in the aqueous humor of age-related macular degeneration (AMD) patients. tRF-1001 upregulation could suppress AMD aqueous humor-induced endothelial sprouting and pathological angiogenesis. Collectively, tRF-1001 acts as an anti-angiogenic factor during ocular angiogenesis. Targeting tRF-1001-mediated signaling is a therapeutic option for ocular neovascular diseases.

8.
Theranostics ; 12(1): 277-289, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34987645

RESUMO

Rationale: Microvascular complication is a major cause of morbidity and mortality among the patients with diabetes. Pericyte dysfunction is the predominant pathological manifestation of microvascular complication. N6-methyladenosine (m6A) serves as the most prevalent modification in eukaryotic mRNAs. However, the role of m6A RNA modification in pericyte dysfunction is still unclear. Methods: Quantitative polymerase chain reactions and western blots were conducted to detect the change of m6A RNA modification in pericytes and mouse retinas following diabetic stress. MTT assay, transwell migration assay, caspase 3/7 activity assay, calcein-AM/propidium iodide (PI) staining, and TUNEL staining were conducted to determine the role of METTL3 in pericyte biology in vitro. Retinal trypsin digestion, vascular permeability assay, and IB4-NG2 double immunofluorescent staining were conducted to determine the role of METTL3 in retinal pericyte dysfunction and vascular complication. RNA sequencing, RNA pull-down assays and immunoblots were conducted to clarify the mechanism of METTL3-mediated pericyte dysfunction and vascular complication. Results: The levels of m6A RNA methylation were significantly up-regulated in pericytes and mouse retinas following diabetic stress, which were caused by increased expression of METTL3. METTL3 regulated the viability, proliferation, and differentiation of pericytes in vitro. Specific depletion of METTL3 in pericytes suppressed diabetes-induced pericyte dysfunction and vascular complication in vivo. METTL3 overexpression impaired pericyte function by repressing PKC-η, FAT4, and PDGFRA expression, which was mediated by YTHDF2-dependent mRNA decay. Conclusion: METTL3-mediated m6A methylation epigenetically regulates diabetes-induced pericyte dysfunction. METTL3-YTHDF2-PKC-η/FAT4/PDGFRA signaling axis could be therapeutically targeted for treating microvascular complications.


Assuntos
Diabetes Mellitus/metabolismo , Retinopatia Diabética/metabolismo , Metiltransferases/metabolismo , Pericitos/metabolismo , Animais , Linhagem Celular , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pericitos/patologia
9.
Mol Med Rep ; 21(6): 2571-2579, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32323773

RESUMO

Ocular angiogenesis is a major cause of severe vision loss, which can affect several parts of the eye, including the retina, choroid and cornea. Vascular endothelial growth factor receptor 2 (VEGFR2) inhibitors have demonstrated great potential for treating ocular angiogenesis and SKLB1002 is a potent inhibitor of VEGF receptor 2 signaling. The present study investigated the effects of SKLB1002 administration on ocular angiogenesis. SKLB1002 administration did not show obvious cytotoxicity and tissue toxicity at the tested concentrations. In an alkali­burn corneal model, SKLB1002 administration significantly decreased the mean length and number of new corneal blood vessels. SKLB1002 administration significantly reduced endothelial cell proliferation, migration and tube formation in vitro. Mechanistically, SKLB1002 inhibited endothelial angiogenic functions by blocking the phosphorylation of ERK1/2, JNK and p38. Thus, selective inhibition of VEGFR­2 through SKLB1002 administration is a promising therapy for ocular angiogenesis.


Assuntos
Neovascularização Fisiológica/efeitos dos fármacos , Quinazolinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Tiadiazóis/farmacologia , Animais , Queimaduras Químicas/metabolismo , Queimaduras Químicas/patologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Córnea/efeitos dos fármacos , Córnea/fisiologia , Doenças da Córnea/metabolismo , Doenças da Córnea/patologia , Modelos Animais de Doenças , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Soluções Oftálmicas/química , Quinazolinas/química , Tiadiazóis/química , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
10.
Zhonghua Nan Ke Xue ; 15(6): 488-92, 2009 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-19593986

RESUMO

OBJECTIVE: To investigate the effects of experimental left varicocele (ELV) on the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) proteins in the testis and epididymis of adolescent rats, and to find out the correlation of the two proteins with varicocele-induced male infertility. METHODS: We established the ELV model in adolescent male SD rats, and detected the expressions of VEGF and Flt-1 proteins in the testis and epididymis by immunohistochemistry at 2 and 4 weeks after surgery. RESULTS: Cell- and region-specific expressions of VEGF and Flt-1 were observed in the testis and epididymis of the ELV and control groups. Statistical analysis showed that, in comparison with the corresponding control groups, the 2- and 4-week ELV groups exhibited a notable increase in the VEGF protein expression in the hibateral testis and epididymis (P < 0.01, P < 0.05); the Flt-1 expression was obviously upregulated in the hibateral testis and epididymis of the 2-week ELV group (P < 0.01, P < 0.01), but remarkably reduced in the hibateral testis and left epididymis of the 4-week ELV group (P < 0.01, P < 0.05), with no statistic difference in the right epididymis (P > 0.05). CONCLUSION: ELV can cause changes in the expressions of VEGF and Flt-1 proteins in the testis and epididymis of adolescent rats, and consequently affect spermatogenesis and spermiotelcosis, which may be one of the causes of varicocele-induced male infertility or subfertility.


Assuntos
Epididimo/metabolismo , Testículo/metabolismo , Varicocele/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Modelos Animais de Doenças , Expressão Gênica , Masculino , Ratos , Ratos Sprague-Dawley
11.
Zhonghua Nan Ke Xue ; 14(3): 200-5, 2008 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-18488329

RESUMO

OBJECTIVE: To investigate the effects of experimental left varicocele (ELV) on the expression of sperm associated antigen 11 (SPAG11) mRNA and its protein isomer SPAG11E in the testis and epididymis of adolescent rats, and to explore the mechanism of infertility caused by varicocele. METHODS: The experimental left varicocele model was established in the adolescent male Sprague-Dawley rats. Two and 4 weeks after the operation, the changes of SPAG11 mRNA and SPAG11E expression in the testis and epididymis were detected using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The expected product of SPAG11 367 bp amplified by RT-PCR was detected only in the epididymis. SPAG11E protein was observed mainly in the acrosomal vesicles and acrosome of round and elongating spermatids of the seminiferous epithelium, in the cytoplasm of Leydig cells, and in the supranuclear region of principle cells and stereocilia of the epididymal epithelium. Imaging and statistical analysis showed that SPAG11 mRNA and SPAG11E protein expressions in the left epididymis of the 2- and 4-week ELV groups presented a remarkable decrease (P < 0.05 or P < 0.01) compared with the right side and the corresponding control group, and the same decreased change in the left epididymis (P < 0.05 or P < 0.01) and an obvious reduction of SPAG11E immunopositive reaction in the right epididymis (P < 0.01) were noted in the 4-week group as compared with the 2-week group. No statistical difference of SPAG11E expression in the bilateral testes was found (P > 0.05) between the ELV group and the control, as well as between the 2- and 4-week ELV groups. CONCLUSION: SPAG11 is a specific gene expressed in the epididymis. The localization and expression of SPAG11E exhibited a region- and cell-specific pattern in both the testis and epididymis of adolescent rats. The expression levels of both SPAG11 mRNA and SPAG11E protein altered obviously in ELV rats. The results suggest that SPAG11 may not only play an important role in spermatogenesis and sperm maturation, but also be associated with varicocele-induced male infertility or subfertility.


Assuntos
Antígenos de Superfície/metabolismo , Epididimo/metabolismo , Glicopeptídeos/metabolismo , Testículo/metabolismo , Varicocele/fisiopatologia , Animais , Antígenos de Superfície/genética , Modelos Animais de Doenças , Glicopeptídeos/genética , Imuno-Histoquímica , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , beta-Defensinas
12.
Zhonghua Nan Ke Xue ; 14(10): 871-5, 2008 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-19157092

RESUMO

OBJECTIVE: To study the expressions of the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) in the testis, epididymis and epididymal sperm of adolescent rats and explore the functions of both the proteins in the male reproductive system. METHODS: The expressions of VEGF and Flt-1 were detected in 20 adolescent SD rats, immunohistochemical staining used for both the testis and the epididymis and immunofluorescent staining for sperm. RESULTS: VEGF and Flt-1 proteins were specifically present in the testis, epididymis and sperm. In the testis, VEGF immunoreactive particles were localized in the cytoplasm of spermatogenic cells, the developing acrosome of spermatids, Sertoli cells and Leydig cells, while Flt-1 expressed mainly in the developing acrosome of spermatids and Leydig cells. In the epididymis, the cell-specific and region-specific expressions of VEGF and Flt-1 proteins were observed in the principal cells of epididymal epithelia, VEGF in the whole epididymis, while Flt-1 only in the caput and cauda segments. Both VEGF and Flt-1 were localized in the acrosome of the sperm head as well as in the neck, middle and principal segments of the sperm tail. CONCLUSION: The specific expression patterns of VEGF and Flt-1 in the rat testis, epididymis and sperm indicate that they may independently or collectively affect spermatogenesis and spermiotelcosis in either an autocrinological or a


Assuntos
Epididimo/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Fator A de Crescimento do Endotélio Vascular/biossíntese , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/biossíntese , Animais , Masculino , Ratos , Ratos Sprague-Dawley , Maturidade Sexual
13.
Asian J Androl ; 9(6): 809-14, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17968467

RESUMO

AIM: To study the incidence of single nucleotide polymorphisms in ubiquitin-specific protease 26 (USP26) gene and its involvement in idiopathic male infertility in China. METHODS: Routine semen analysis was performed. Infertility factors such as immunological, infectious and biochemical disorders were examined to select patients with idiopathic infertility. DNA was isolated from peripheral blood of the selected patients and control population, which were examined for mutations using polymerase chain reaction-single strand conformation polymorphism analysis. Furthermore, nucleotide sequences were sequenced in some patients and controls. RESULTS: Of 41 infertile men, 9 (22.0%, P = 0.01) had changes in USP26 gene on the X chromosome. A compound mutation (364insACA; 460G right triple arrow A) was detected in 8 patients (19.5%, P = 0.01) and a 1044T right triple arrow A substitution was found in 1 patient (2.4%, P > 0.05). All three variations led to changes in the coding amino acids. Two substitutions predict some changes: 460G right triple arrow A changes a valine into an isoleucine, and 1044T right triple arrow A substitutes a leucine for a phenylalanine. Another insertion of three nucleotides ACA causes an insertion of threonine. No other changes were found in the remaining patients and fertile controls. CONCLUSION: The USP26 gene might be of importance in male reproduction. Mutations in this gene might be associated with male infertility, and might negatively affect testicular function. Further research on this issue is in progress.


Assuntos
Endopeptidases/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único/genética , Espermatogênese/genética , Povo Asiático/etnologia , Povo Asiático/genética , Estudos de Casos e Controles , China , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Endopeptidases/metabolismo , Humanos , Incidência , Infertilidade Masculina/etnologia , Células Intersticiais do Testículo/metabolismo , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Células de Sertoli/metabolismo , Testículo/metabolismo , Proteases Específicas de Ubiquitina
14.
Chin Med J (Engl) ; 130(21): 2585-2590, 2017 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-29067957

RESUMO

BACKGROUND: Intermittent hypoxia (IH) is a key element of obstructive sleep apnea (OSA) that can lead to disorders in the liver. In this study, IH was established in a rat model to examine its effects on the expression of hepatic cytochrome P450 (CYP) and CYP regulators, including nuclear receptors. METHODS: Hematoxylin and eosin staining was conducted to analyze the general pathology of the liver of rats exposed to IH. The messenger RNA (mRNA) expression levels of inflammatory cytokines, CYPs, nuclear factor-κB (NF-κB), and nuclear factors in the liver were measured by quantitative reverse transcription polymerase chain reaction. RESULTS: We found inflammatory infiltrates in the liver of rats exposed to IH. The mRNA expression level of interleukin-1beta was increased in the liver of the IH-exposed rats (0.005 ± 0.001 vs. 0.038 ± 0.008, P = 0.042), whereas the mRNA expression level of Cyp1a2 was downregulated (0.022 ± 0.002 vs. 0.0050 ± 0.0002, P = 0.029). The hepatic level of transcription factor NF-κB was also reduced in the IH group relative to that in the control group, but the difference was not statistically significant and was parallel to the expression of the pregnane X receptor and constitutive androstane receptor. However, the decreased expression of the glucocorticoid receptor upon IH treatment was statistically significant (0.056 ± 0.012 vs. 0.032 ± 0.005, P = 0.035). CONCLUSIONS: These results indicate a decrease in expression of hepatic CYPs and their regulator GR in rats exposed to IH. Therefore, this should be noted for patients on medication, especially those on drugs metabolized via the hepatic system, and close attention should be paid to the liver function of patients with OSA-associated IH.


Assuntos
Citocromo P-450 CYP1A2/metabolismo , Hipóxia/metabolismo , Inflamação/metabolismo , Fígado/metabolismo , Animais , Receptor Constitutivo de Androstano , Masculino , Receptor de Pregnano X , Ratos , Ratos Wistar , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores de Glucocorticoides/metabolismo , Receptores de Esteroides/metabolismo
15.
Zhonghua Nan Ke Xue ; 12(9): 780-3, 2006 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-17009525

RESUMO

OBJECTIVE: To investigate the expression of human epididymal secretary protein 2 isoform human epididymal protein 2beta1(HE2beta1) in the testis and epididymis of adolescent male rats along with its significance. METHODS: Immunohistochemical staining was used to detect the expression and localization of HE2beta1 in the testis and epididymis of 15 adolescent SD rats. RESULTS: HE2beta1 immunoreactive staining was detected in the testis and epididymis. In the epithelia of the epididymal duct, HE2beta1 expressed mainly in the supranuclear region of the principle cells and the basement membrane of some epithelial cells; there were no immunostaining in the n clear cells, halo cells and basal cells. The immunopositive reaction was detected, weak in the distal caput, strong in the proximal, middle corpus and the cauda, but negative in the initial segment. Immunopositive results of HE2beta1 were also observed in some of the nuclei of spermatogonia and Sertoli cells with negatively-stained cytoplasm. CONCLUSION: Immunohistochemical staining is a fairly sensitive method for detecting HE2beta1 expression. The localization and expression level of HE2beta1 in the genital duct of adolescent male rats exhibited a region- and cell-specific expression pattern, which suggests that HE2beta1 may play an important role in spermatogenesis, maturation and epididymal epithelial innate defense mechanisms.


Assuntos
Antígenos de Superfície/biossíntese , Epididimo/metabolismo , Glicopeptídeos/biossíntese , Testículo/metabolismo , Animais , Humanos , Imuno-Histoquímica , Células Intersticiais do Testículo/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley
16.
Zhonghua Nan Ke Xue ; 12(11): 974-8, 2006 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-17146919

RESUMO

OBJECTIVE: To investigate the effects of experimental left varicocele (ELV) on the cystatin-related epididymal spermatogenic (CRES) protein in the testis and epididymis of adolescent rats. METHODS: The ELV model of Sprague-Dawley (SD) male adolescent rats was established, and the expression of CRES protein in the testis and epididymis was detected by immunohistochemistry and Western-blot at 2 and 4 weeks after surgery. RESULTS: Immunohistochemistry and Western-blot detected CRES protein in both the testis and the epididymis of the ELV rats and the control rats. Immunohistochemistry showed that within the testis, CRES protein was expressed mainly in the cytoplasm of round spermatids and elongating spermatids, sperm acrosomes and residual bodies. The expression was most intensive at Stages I-III and IX-XIV, and then decreased gradually at Stages VII-VII and IV-VI. Within the epididymis, CRES protein was expressed mainly in the cytoplasm of the principal cells of epididymal epithelia. Western-blot detected CRES protein in Mr 19,000 and 14,000, stronger in the former than in the latter. Image and statistical analyses showed that the expression of CRES protein in the 2-week and 4-week ELV groups was significantly higher than in the control group (P < 0.05, or P < 0.01). CONCLUSION: CRES protein expressed in both the testis and epididymis of adolescent rats and the expression is stage-specific and cell-specific in the testis and segment-specific and cell-specific in the epididymis. The expression of CRES protein in the ELV rats is much stronger than in their corresponding controls. It is suggested that CRES protein may be significantly involved in the regulation of spermatogenesis and sperm maturation, and possibly associated with varicocele-related male infertility or subfertility.


Assuntos
Cistatinas/biossíntese , Epididimo/metabolismo , Testículo/metabolismo , Varicocele/metabolismo , Animais , Western Blotting , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Ratos , Ratos Sprague-Dawley
17.
Phytomedicine ; 23(5): 468-76, 2016 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-27064005

RESUMO

BACKGROUND: Epidermal growth factor receptor (EGFR) is an effective molecular target for cancer treatment. Boehmenan, a lignan from the dried stems of Clematis armandii, exhibited the potent cytotoxic effects against many cancer cell lines in previous studies. However, the effects and underlying mechanism of boehmenan on non-small cell lung cancer (NSCLC) remains unclear. PURPOSE: The present study was designed to determine the in vitro anti-cancer properties and underlying molecular mechanisms of boehmenan on A549 NSCLC cells. STUDY DESIGN/METHODS: Cellular viability and chemoattractive properties of macrophages were investigated by using MTT and transwell migration assay, respectively. Mitochondrial membrane potential (ΔΨm), apoptotic ratio, and cell cycle were measured by flow cytometry. Protein expression was visualized by Western blot using specific antibodies. RESULTS: Boehmenan concentration-dependently suppressed proliferation and induced G1 phase arrest in A549 NSCLC cells, which were accompanied by reduction of migration, colony formation and increase of apoptosis in A549 cells. In addition, boehmenan treatment markedly modulated apoptosis-related protein (p53, p21, cleaved caspase 3, and cleaved PARP) and cyclin D1 expression and induced ΔΨm collapse in a concentration dependent manner. Furthermore, boehmenan concentration-dependently inhibited EGF-induced activation of EGFR and its downstream signaling molecules, including MEK, Akt, ERK1/2, and STAT3. CONCLUSION: Taken together, our results suggested that boehmenan-mediated anti-tumor property was mediated by modulation of mitochondria and EGFR signaling pathway in A549 NSCLC cells.


Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/patologia , Clematis/química , Fator de Crescimento Epidérmico/metabolismo , Lignanas/farmacologia , Neoplasias Pulmonares/patologia , Caspase 3/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Receptores ErbB/metabolismo , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Estrutura Molecular , Plantas Medicinais/química , Transdução de Sinais/efeitos dos fármacos
18.
Phytomedicine ; 23(5): 541-9, 2016 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-27064013

RESUMO

BACKGROUND: (7R, 8S)-9-Acetyl-dehydrodiconiferyl alcohol (ADDA), a novel lignan compound isolated from Clematis armandii Franch (Ranunculaceae) stems, has been found to exert potential anti-inflammatory activities in vitro. PURPOSE: To investigate the pharmacological effects and molecular mechanisms of ADDA on lipopolysaccharide (LPS)-induced activation and migration of macrophages. STUDY DESIGN/METHODS: Macrophages were stimulated with LPS in the presence or absence of ADDA. Expression of inflammatory mediators, including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nitric oxide (NO) were measured by Western blot and commercial NO detection kit. Cellular viability and chemotactic properties of macrophages were investigated using MTT and transwell migration assays. The activation and expression of mitogen activated protein kinases, nuclear factor-κB (NF-κB), protein kinase B (Akt), Src, and focal adhesion kinase (FAK) were analyzed by Western blot. RESULTS: Non-toxic concentrations (12.5-50 µM) of ADDA concentration-dependently inhibited expression/release of inflammatory mediators (COX-2, iNOS, and NO), suppressed Akt and c-jun N-terminal kinase 1/2 (JNK) phosphorylation, and NF-κB activation in LPS-stimulated macrophages. In addition, ADDA blocked LPS-mediated macrophage migration and this was associated with inhibition of LPS-induced Src and FAK phosphorylation as well as Src expression in a concentration dependent manner. Notably, the inhibitory effects of ADDA on iNOS, NO, and Src could be mimicked by a Src inhibitor PP2 or an iNOS inhibitor l-NMMA. CONCLUSION: Our results suggested that ADDA attenuated LPS-induced inflammatory responses in macrophages and cell migration, at least in part, through inhibition of NF-κB activation and modulation of iNOS/Src/FAK axis.


Assuntos
Anti-Inflamatórios/farmacologia , Lignanas/farmacologia , Macrófagos/efeitos dos fármacos , Animais , Movimento Celular/efeitos dos fármacos , Clematis/química , Ciclo-Oxigenase 2/metabolismo , Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Proteína Quinase 8 Ativada por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Caules de Planta/química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células RAW 264.7
19.
Cell Biochem Biophys ; 72(1): 83-7, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25416584

RESUMO

The objective of this study was to examine the clinical effectiveness of scapulothoracic joint control training exercises on shoulder joint dysfunction. Forty patients with traumatic shoulder pain and joint dysfunction were randomized into the treatment or control group. Standard rehabilitation interventions included glenohumeral joint mobilization techniques, ultrasound therapy, traditional Chinese medicine, interference current therapy, and other comprehensive interventions. Patients received scapulothoracic joint control training exercises, including active and passive motions of the scapulothoracic joints, peri-joint muscle exercise, and joint stability exercises for 1 month. Patient status was evaluated by Constant-Murley scales before and after the prescribed interventions. The pain conditions, daily activities, range of movement, strength tests and total scores were significantly improved compared to prior treatment. Moreover, improvements in pain, daily activities, scope of activities, and total scores for patients in the treatment group were statistically significant when compared to the control group (P < 0.05). However, there was no inter-group difference in strength testing. The combination of standard rehabilitation interventions and scapulothoracic joint control training exercises are an effective treatment of the shoulder joint dysfunction. Moreover, the pain outcomes, scope of activities, and total scores were better in the treatment group.


Assuntos
Terapia por Exercício/métodos , Amplitude de Movimento Articular , Lesões do Ombro , Articulação do Ombro/fisiopatologia , Dor de Ombro/terapia , Adulto , Eletrodos , Feminino , Humanos , Masculino , Medicina Tradicional Chinesa , Pessoa de Meia-Idade , Movimento , Reabilitação/métodos , Escápula/lesões , Escápula/fisiopatologia , Resultado do Tratamento
20.
Asian J Androl ; 5(2): 108-12, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12778320

RESUMO

AIM: To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats. METHODS: ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed. RESULTS: In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05). CONCLUSION: There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.


Assuntos
Epididimo/patologia , Epididimo/fisiopatologia , Varicocele/patologia , Varicocele/fisiopatologia , Animais , Carnitina/metabolismo , Epididimo/enzimologia , Epididimo/ultraestrutura , Masculino , Microscopia Eletrônica , Ratos , Ratos Sprague-Dawley , Varicocele/enzimologia , alfa-Glucosidases/metabolismo
SELEÇÃO DE REFERÊNCIAS
Detalhe da pesquisa