RESUMO
Inflammation of cartilage is a primary symptom for knee-joint osteoarthritis. Matrix metalloproteinases (MMPs) are known to play an important role in the articular cartilage destruction related to osteoarthritis. Naringenin is a plant-derived flavonoid known for its anti-inflammatory properties. We studied the effect of naringenin on the transcriptional expression, secretion and enzymatic activity of MMP-3 in vivo in the murine monosodium iodoacetate (MIA) osteoarthritis model. The assessment of pain behavior was also performed in the MIA rats. The destruction of knee-joint tissues was analyzed microscopically. Moreover, the effect of naringenin was also studied in vitro in IL-1ß activated articular chondrocytes. The transcriptional expression of MMP-3, MMP-1, MMP-13, thrombospondin motifs (ADAMTS-4) and ADAMTS-5 was also studied in primary cultured chondrocytes of rats. Naringenin caused significant reduction in pain behavior and showed marked improvement in the tissue morphology of MIA rats. Moreover, a significant inhibition of MMP-3 expression in MIA rats was observed upon treatment with naringenin. In the in vitro tests, naringenin caused a significant reduction in the transcriptional expression, secretion and enzymatic activity of the studied degradative enzymes. The NF-κB pathway was also found to be inhibited upon treatment with naringenin in vitro. Overall, the study suggests that naringenin alleviated pain and regulated the production of matrix-metalloproteinases via regulation of NF-κB pathway. Thus, naringenin could be a potent therapeutic option for the treatment of osteoarthritis.
Assuntos
Anti-Inflamatórios/farmacologia , Artralgia/enzimologia , Condrócitos/enzimologia , Flavanonas/farmacologia , Articulação do Joelho/enzimologia , Metaloproteinase 3 da Matriz/biossíntese , Osteoartrite do Joelho/enzimologia , Animais , Artralgia/tratamento farmacológico , Western Blotting , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Condrócitos/efeitos dos fármacos , Modelos Animais de Doenças , Expressão Gênica , Interleucina-1beta/análise , Interleucina-1beta/efeitos dos fármacos , Interleucina-1beta/metabolismo , Articulação do Joelho/patologia , Masculino , Metaloproteinase 3 da Matriz/análise , Inibidor de NF-kappaB alfa/análise , Inibidor de NF-kappaB alfa/efeitos dos fármacos , NF-kappa B/análise , NF-kappa B/efeitos dos fármacos , Osteoartrite do Joelho/tratamento farmacológico , Osteoartrite do Joelho/patologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Resultado do TratamentoRESUMO
We investigated the association between serum visfatin levels and single nucleotide polymorphisms (SNPs; rs61330082, rs2058539) in the visfatin gene and coronary artery calcification (CAC) in patients from Wenzhou, China. CAC patients (N = 206) were divided into two groups: mild CAC (MCAC) and moderate and severe CAC (MSCAC). Volunteers without CAC (N = 70) were included in the control group. The serum visfatin level was analyzed by enzyme-linked immunosorbent assay. SNPs (rs61330082, rs2058539) in the visfatin gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Clinical data, serum visfatin levels, and genotype and allele frequencies of rs61330082 and rs2058539 were compared among the three groups. MSCAC patients expressed significantly higher serum visfatin levels (30.58 ± 6.12 ng/mL) than individuals in the MCAC (29.03 ± 1.87 ng/mL) and control (24.45 ± 5.44 ng/mL) groups (P < 0.05). The genotype distributions and frequencies of rs61330082 differed significantly among the groups (P < 0.05), while those of rs2058539 did not. The serum visfatin level was positively correlated with the body mass index (BMI), high-density lipoprotein cholesterol (HDL-C), and insulin resistance index (IRI), and negatively correlated with the triglyceride (TG) levels (P < 0.05) of patients. Serum visfatin is associated with the development of CAC. The T allele of the rs61330082 SNP in the visfatin gene had a cardioprotective effect on patients with CAC; the SNP at rs2058539 was not significantly associated with CAC. The BMI, HDL-C, IRI, and TG levels influenced the development of CAC.
Assuntos
Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/genética , Citocinas/sangue , Citocinas/genética , Nicotinamida Fosforribosiltransferase/sangue , Nicotinamida Fosforribosiltransferase/genética , Calcificação Vascular/sangue , Calcificação Vascular/genética , Idoso , Alelos , Povo Asiático/genética , HDL-Colesterol/sangue , Vasos Coronários/fisiopatologia , Citocinas/biossíntese , Feminino , Frequência do Gene , Genótipo , Humanos , Insulina/sangue , Insulina/genética , Masculino , Pessoa de Meia-Idade , Nicotinamida Fosforribosiltransferase/biossíntese , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The association between the HLA-DP single nucleotide polymorphisms (SNPs) rs3077 and rs9277535 and hepatocellular carcinoma (HCC) has been reported, but results have been inconclusive and controversial. Therefore, to investigate the relationship between these HLA-DP SNPs and HCC susceptibility, a meta-analysis of studies published before January 2014 was carried out using the PubMed and Google Scholar databases. Odds ratios (ORs) and 95% confidence intervals (CI) were calculated for HLA-DP alleles, and for co-dominant, dominant, and recessive genotype models of each SNP, based on fixed- or random-effects models. A total of nine studies from six published articles were included. The association study between rs3077 and HCC susceptibility was performed in four independent comparisons that contained 1871 cases with hepatitis B virus (HBV)-related HCC and 3207 carriers with persistent HBV. Association between rs9277535 and HCC susceptibility was examined in five separate comparisons that contained 2017 cases and 3930 carriers. Our analysis indicated a significant association of rs3077 and rs9277535 with HCC susceptibility, suggesting that rs3077 might act beneficially against HCC susceptibility (A vs G: OR = 0.884, 95%CI = 0.803-0.973, P = 0.012; GA vs GG: OR = 0.842, 95%CI = 0.733-0.967, P = 0.015; AA+GA vs GG: OR = 0.848, 95%CI = 0.744-0.968, P = 0.014), and that rs9277535 might promote HCC susceptibility (AA vs GA: OR = 1.202, 95%CI = 1.011-1.428, P = 0.037). This study suggested that HLA-DP rs3077 and rs9277535 polymorphisms are associated with HCC susceptibility in the Asian population.
Assuntos
Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/genética , Predisposição Genética para Doença , Antígenos HLA-DP , Neoplasias Hepáticas/epidemiologia , Neoplasias Hepáticas/genética , Polimorfismo de Nucleotídeo Único , Alelos , Povo Asiático/genética , Estudos de Casos e Controles , Frequência do Gene , Estudos de Associação Genética , Haplótipos , Humanos , Razão de Chances , Viés de Publicação , RiscoRESUMO
We constructed a plasmid containing a protein transduction domain (PTD) and a human A20 (hA20) gene fragment; the fusion protein was obtained by highly expressing this plasmid in the yeast Pichia pastoris GS115. The plasmid was obtained by adding 9xArg and EcoRÐ recognition sites to the end of the primer, and 6xHis-Tag and NotÐ recognition sites to its end. After sequencing, the hA20 gene fragment was inserted into plasmid pPIC9k to construct expression vector pPIC9k-PTD-hA20; then, we transfected GS115 with the vector and induced PTD-hA20 protein expression. We purified protein from the yeast fermentation supernatant using a nickel column. Human umbilical vein endothelial cells (HUVECs) were cultured in high glucose medium (30 mM glucose) and in high glucose medium containing different concentrations of protein. Apoptosis of HUVECs was assayed by TUNEL 72 h later. The biological activity tests indicated that the fusion protein not only passed through the cell membrane freely, but also inhibited apoptosis of HUVECs induced by high glucose levels. We conclude that the fusion protein PTD-hA20 has potential for clinical use.
Assuntos
Citoproteção/efeitos dos fármacos , Proteínas de Ligação a DNA/farmacologia , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Glucose/toxicidade , Peptídeos e Proteínas de Sinalização Intracelular/farmacologia , Proteínas Nucleares/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Eletroforese em Gel de Ágar , Células Endoteliais/metabolismo , Genoma Fúngico/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Microscopia de Fluorescência , Mutagênese Insercional/genética , Pichia/efeitos dos fármacos , Pichia/genética , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/isolamento & purificação , Recombinação Genética/genética , Proteína 3 Induzida por Fator de Necrose Tumoral alfaRESUMO
Diabetes mellitus causes vascular lesions and may ultimately lead to atherosclerosis. One of the earliest steps in the development of atherosclerotic lesions is the adhesion of monocytes to endothelial cells of the vessel wall. It is currently unknown whether zinc finger protein A20 is able to protect endothelial cells from injury caused by high levels of glucose and monocyte homing. In our study, adhesion of monocytes to the vessel wall endothelium was detected by measuring the rolling velocity of monocytes along human umbilical vein endothelial cells (HUVECs). Activation of NF-κB was analyzed through Western blot. HUVEC apoptosis was monitored by TUNEL in situ end-labeling and flow cytometry. High glucose concentrations (25 mM) stimulated monocytes, reducing the velocity at which they roll along HUVECs. Stimulation of monocytes with high levels of glucose also induced HUVEC apoptosis. Overexpression of the zinc finger protein A20 inhibited monocyte recruitment, NF-κB activation, P-selectin expression, and HUVEC apoptosis induced by high glucose levels. We conclude that zinc finger protein A20 can protect HUVECs from injury induced by high levels of glucose and potentially could be used to develop treatments against diabetic vascular lesions.