ABSTRACT
We report the changed levels of serum amyloid alpha, an immunologically active protein, in Parkinson's disease (PD) patients' peripheral tissues. We have previously shown that Saa-1 and -2 (serum amyloid alpha-1,-2, genes) were among the top downregulated genes in PD patients' skin, using whole-genome RNA sequencing. In the current study, we characterized the gene and protein expression profiles of skin and blood samples from patients with confirmed PD diagnosis and age/sex matched controls. qRT-PCR analysis of PD skin demonstrated downregulation of Saa-1 and -2 genes in PD patients. However, the lowered amount of protein could not be visualized using immunohistochemistry, due to low quantity of SAA (Serum Amyloid Alpha, protein) in skin. Saa-1 and -2 expression levels in whole blood were below detection threshold based on RNA sequencing, however significantly lowered protein levels of SAA1/2 in PD patients' serum were shown with ELISA, implying that SAA is secreted into the blood. These results show that SAA is differentially expressed in the peripheral tissues of PD patients.
Subject(s)
Interleukin-1/genetics , Interleukins/genetics , Psoriasis/genetics , Case-Control Studies , Databases, Genetic , Gene Expression Regulation , Humans , Immunohistochemistry , Interleukin-1/analysis , Interleukins/analysis , Psoriasis/diagnosis , Psoriasis/immunology , RNA, Messenger/genetics , Sequence Analysis, RNAABSTRACT
BACKGROUND: Chronic skin inflammation in atopic dermatitis (AD) is associated with elevated expression of proinflammatory genes and activation of innate immune responses in keratinocytes. microRNAs (miRNAs) are short, single-stranded RNA molecules that silence genes via the degradation of target mRNAs or inhibition of translation. OBJECTIVE: The aim of this study was to investigate the role of miR-146a in skin inflammation in AD. METHODS: RNA and protein expression was analyzed using miRNA and mRNA arrays, RT-quantitative PCR, Western blotting, and immunonohistochemistry. Transfection of miR-146a precursors and inhibitors into human primary keratinocytes, luciferase assays, and MC903-dependent mouse model of AD were used to study miR-146a function. RESULTS: We show that miR-146a expression is increased in keratinocytes and chronic lesional skin of patients with AD. miR-146a inhibited the expression of numerous proinflammatory factors, including IFN-γ-inducible and AD-associated genes CCL5, CCL8, and ubiquitin D (UBD) in human primary keratinocytes stimulated with IFN-γ, TNF-α, or IL-1ß. In a mouse model of AD, miR-146a-deficient mice developed stronger inflammation characterized by increased accumulation of infiltrating cells in the dermis, elevated expression of IFN-γ, CCL5, CCL8, and UBD in the skin, and IFN-γ, IL-1ß, and UBD in draining lymph nodes. Both tissue culture and in vivo experiments in mice demonstrated that miR-146a-mediated suppression in allergic skin inflammation partially occurs through direct targeting of upstream nuclear factor kappa B signal transducers caspase recruitment domain-containing protein 10 and IL-1 receptor-associated kinase 1. In addition, human CCL5 was determined as a novel, direct target of miR-146a. CONCLUSION: Our data demonstrate that miR-146a controls nuclear factor kappa B-dependent inflammatory responses in keratinocytes and chronic skin inflammation in AD.
Subject(s)
Dermatitis, Atopic/genetics , Keratinocytes/immunology , MicroRNAs/physiology , NF-kappa B/metabolism , RNA Interference , Skin/immunology , Animals , Calcitriol/administration & dosage , Calcitriol/analogs & derivatives , Cell Movement/genetics , Cells, Cultured , Chronic Disease , Cytokines/genetics , Cytokines/immunology , Cytokines/metabolism , Dermatitis, Atopic/chemically induced , Dermatitis, Atopic/immunology , Disease Models, Animal , Humans , Immunity, Innate , Immunosuppression Therapy , Inflammation/genetics , Inflammation Mediators/metabolism , Mice , Mice, Inbred C57BL , MicroRNAs/genetics , NF-kappa B/genetics , RNA Interference/immunology , Signal Transduction/genetics , Skin/pathology , Up-RegulationABSTRACT
OBJECTIVE: To compare markers of semen quality and related reproductive indicators in middle-aged males in relation to serum prostate-specific antigen (PSA) levels. METHODS: A total of 384 voluntary middle-aged men who underwent screening for prostate health were recruited. Reproductive function, semen quality, hormonal parameters, prostate-related pathologies, and PSA levels were measured. RESULTS: Total sperm count and sperm concentration were decreased in men with elevated serum PSA compared with age-matched controls. Serum FSH levels differed significantly in subjects with non-malignant, pre-malignant, and malignant prostate conditions. PSA levels in men with three normal semen variables (semen volume ≥ 1.5 mL, sperm density ≥ 15 × 10(6)/mL, and A + B motility ≥ 40%) were significantly lower compared with subjects with altered parameters (1.51 ng/ml vs. 2.47 ng/ml, respectively, p = 0.002). PSA showed a negative correlation with semen volume and total sperm count, and a positive correlation with semen interleukin-6. CONCLUSIONS: Our data demonstrate that serum PSA levels are related to impaired reproductive parameters in middle-aged subjects. While there is no consensus about the potential link between male factor infertility, impaired reproductive indicators, and prostate pathologies, this topic needs additional research focusing on the interactions between male aging, reproductive function, and the prostate.
