RESUMEN
Ballooning degeneration (BD) of hepatocytes is a distinguishing histological feature associated with the progression of nonalcoholic fatty liver disease (NAFLD). Under the assumption that NAFLD severity is associated with metabolic-stress we explored the hypothesis that heat shock 27 kDa protein 1 (HSP27), a protein chaperone involved in stress resistance and cytoskeletal-remodeling, might be deregulated in ballooned hepatocytes. We observed that fasting plasma glucose (fpG) (p = 0.00002), total cholesterol (p = 0.02) and triglycerides (p = 0.01) levels, and female sex (p = 0.01) were significantly associated with the presence of BD. A logistic regression model showed that BD was independently associated with fpG (p = 0.002); OR per unit of glucose concentration 1.05, 95% confidence interval 1.02-1.09. Furthermore, BD was associated with a significant 2.24-fold decrease in the expression level of HSP27-mRNA in comparison with absence of ballooning, p = 0.002. Ballooned hepatocytes showed very low HSP27 immunoreactivity compared with hepatocyes without ballooning (p = 0.009); HSP27 immunoreactivity was inversely correlated with fpG levels (R: -0.49, p = 0.01). In conclusion, BD is associated with down-regulation of liver HSP27 gene and protein expression, suggesting that ballooned hepatocytes fail to ensure a robust physiological response to metabolic-induced stress.
Asunto(s)
Regulación hacia Abajo , Proteínas de Choque Térmico HSP27/biosíntesis , Hepatocitos/metabolismo , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Adulto , Femenino , Proteínas de Choque Térmico , Hepatocitos/patología , Humanos , Masculino , Persona de Mediana Edad , Chaperonas Moleculares , Enfermedad del Hígado Graso no Alcohólico/patología , Enfermedad del Hígado Graso no Alcohólico/terapiaRESUMEN
Suboptimal intake of Zinc (Zn) is one of the most common worldwide nutritional problems. The aim of this study is to provide new evidence on the relation between moderate Zn restriction, and cytoprotective functions in airway epithelium. We analyzed the effect of moderate Zn deficiency (ZD) on the expression of several pro and anti-apoptotic proteins and cytoprotective factors (Hsp27 and Hsp 70i), as well as the effect of restoring Zn during the refeeding period. Adult male rats were divided into three groups: Zn-adequate control group, Zn-deficient group and Zn-refed group. Our previous findings showed an important oxidative and nitrosative stress during ZD, this situation is accompanied by inflammation and alterations in the expression of matrix extracellular proteins. We observed a strong immunopositive area of anti and pro-apoptotics proteins in ZD groups. The mRNA levels of Nrf-2, Bax and Bad were increased in ZD, while in ZD refed group its levels were similar to the control values. The increased expression of Nrf-2 is likely to be critical for protection of lung under inflammatory process triggered during ZD. Hsp27 and Hsp 70i showed an increase of immunostaining area but they were not significant. During the supplementation period, heat-shock proteins increased significantly. In conclusion, our results provide further evidence of the pathways involved in cytoprotection and apoptosis caused by ZD. Additional studies are required in order to investigate whether Hsp27 and Hsp70 are consistently associated with cellular stress and inflammation in lung. There may be a beneficial role for improved Zn nutrition or Zn supplements early in lung pathology.
Asunto(s)
Citoprotección , Células Epiteliales/citología , Pulmón/citología , Zinc/deficiencia , Animales , Apoptosis/efectos de los fármacos , Citoprotección/genética , Dieta , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Proteínas de Choque Térmico HSP27/análisis , Proteínas de Choque Térmico HSP27/biosíntesis , Proteínas HSP70 de Choque Térmico/análisis , Proteínas HSP70 de Choque Térmico/biosíntesis , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Masculino , Ratas , Ratas Wistar , Zinc/administración & dosificación , Zinc/farmacologíaRESUMEN
Cystic ovarian disease (COD), which is considered one of the most important causes of reproductive failure in dairy cattle, induces intraovarian changes in the expression of numerous genes. The purpose of this study was to analyze the changes in the expression of Heat Shock Proteins (HSPs) in ovaries from bovines with cystic ovarian disease induced by ACTH. Immunoreactivity for Heat Shock Proteins (HSPs) in ovaries of cows with induced COD showed differential expression patterns in growing follicles from the control group. The immunopositive area for Hsp27 and Hsp60 in granulosa cells showed significant differences between tertiary follicles from normal cycling animals and those from animals with induced COD. The cysts showed increased Hsp27 immunostaining in theca cells in relation to tertiary follicles from normal cycling cows. Hsp70 immunostaining was more intense in cystic follicles than in other follicular categories from animals with induced COD, in both granulosa and theca cells. In granulosa cells, tertiary follicles from the control group showed higher levels of Hsp90 than cysts. These results demonstrate that there are differences in HSP protein expression when COD is induced. In fact, HSP expression would be part of the functional response to the changes in hormones and neurotransmitters induced by stress, indicating that HSPs can control hormonal functions and vice versa.
Asunto(s)
Hormona Adrenocorticotrópica/farmacología , Enfermedades de los Bovinos/metabolismo , Proteínas de Choque Térmico/biosíntesis , Quistes Ováricos/veterinaria , Ovario/metabolismo , Animales , Bovinos , Enfermedades de los Bovinos/inducido químicamente , Chaperonina 60/biosíntesis , Chaperonina 60/fisiología , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Células de la Granulosa/efectos de los fármacos , Células de la Granulosa/metabolismo , Proteínas de Choque Térmico HSP27/biosíntesis , Proteínas de Choque Térmico HSP27/fisiología , Proteínas de Choque Térmico/fisiología , Quistes Ováricos/inducido químicamente , Quistes Ováricos/metabolismo , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/metabolismo , Ovario/efectos de los fármacos , Células Tecales/efectos de los fármacos , Células Tecales/metabolismoRESUMEN
We investigated in vitro the effect of low-frequency and low-energy ultrasound (LFLEU) on apoptosis of an overexpressed HSP27 human aortic smooth muscle cell (HASMC) line. A frequency of 42.6 kHz was used in all experiments. HASMC were exposed to ultrasound and cell viability was evaluated by MTT reduction. Overexpressed HSP27-HASMC was constructed on a pcDNA3.1 vector. Apoptosis was determined 24 h after treatment by flow cytometry; gene display was evaluated with Affimax chips, and HSP27 mRNA and protein expression levels were measured by RT-PCR and Western blotting. The apoptosis rate (at 30 s) was significantly lower in HASMC transfected with HSP27 (7.14 ± 1.73%), compared with cells transfected with a mock plasmid (17.31 ± 2.72%) or a control group (14.23 ± 2.77%), indicating a protective function for apoptosis induced by LFLEU. Gene display analysis showed that caspase-9 expression in HSP27 cell lines was downregulated and caspase-3 upregulated. However, RT-PCR and Western blotting analysis indicated that both caspase-9 and caspase-3 were inhibited at both the mRNA and protein levels. We suggest that overexpressed HSP27 is capable of protecting the LFLEU from apoptosis and that the pathway for this protection is via downregulated caspase-9 and caspase-3 expression.