Proteomic Fingerprint of Lung Fibrosis Progression and Response to Therapy in Bleomycin-Induced Mouse Model

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by the aberrant accumulation of extracellular matrix in the lungs. nintedanib is one of the two FDA-approved drugs for IPF treatment; however, the exact pathophysiological mechanisms of fibrosis progression and response to therapy are still poorly understood. In this work, the molecular fingerprint of fibrosis progression and response to nintedanib treatment have been investigated by mass spectrometry-based bottom-up proteomics in paraffin-embedded lung tissues from bleomycin-induced (BLM) pulmonary fibrosis mice. Our proteomics results unveiled that (i) samples clustered depending on the tissue fibrotic grade (mild, moderate, and severe) and not on the time course after BLM treatment; (ii) the dysregulation of different pathways involved in fibrosis progression such as the complement coagulation cascades, advanced glycation end products (AGEs) and their receptors (RAGEs) signaling, the extracellular matrix-receptor interaction, the regulation of actin cytoskeleton, and ribosomes; (iii) Coronin 1A (Coro1a) as the protein with the highest correlation when evaluating the progression of fibrosis, with an increased expression from mild to severe fibrosis; and (iv) a total of 10 differentially expressed proteins (padj-value ≤ 0.05 and Fold change ≤-1.5 or ≥1.5), whose abundance varied in the base of the severity of fibrosis (mild and moderate), were modulated by the antifibrotic treatment with nintedanib, reverting their trend. Notably, nintedanib significantly restored lactate dehydrogenase B (Ldhb) expression but not lactate dehydrogenase A (Ldha). Notwithstanding the need for further investigations to validate the roles of both Coro1a and Ldhb, our findings provide an extensive proteomic characterization with a strong relationship with histomorphometric measurements. These results unveil some biological processes in pulmonary fibrosis and drug-mediated fibrosis therapy

A solid-phase radioimmunoassay for IgG gliadin antibodies using 125I-labelled staphylococcal protein A.

A sensitive radioimmunoassay for IgG gliadin antibodies is described. Serum specimens were added to wells of plastic microtitre plates coated with gliadin. After removal of the unbound material, gliadin antibodies were detected by adding 125I-labelled staphylococcal protein A (125I-SpA). Serum specimens from coeliac patients on a normal diet or on a gluten-free diet were tested, as well as sera from an age-matched control group. Measurements to obtain precise quantitative values were made with gliadin antibody-rich serum as reference standard. High titres of gliadin antibodies were found in 18 out of 19 coeliac patients on a normal diet (95%); in patients on a strict gluten-free diet serum values did not exceed 2 S.D. of the control mean. Due to the high sensitivity of the method a low but detectable amount of gliadin antibody was present in the sera of all controls

A potential physiological involvement of nesfatin-1 in regulating swine granulosa cell functions

Nesfatin-1 has recently been indicated as a pleiotropic molecule, mainly involved in the metabolic regulation of reproductive functions acting at hypothalamic level. The aim of the present study was to explore a local action of nesfatin-1 in swine ovarian follicle. Firstly, the expression of NUCB-2 by qRT-PCR was verified in swine granulosa cells from different size follicles and nesfatin-1 was localized by immunohistochemistry in sections of the whole porcine ovary. In addition, the in vitro effect of different nesfatin-1 concentrations was tested on cell growth, steroidogenesis and redox status of granulosa cells. Finally, the effect of nesfatin-1 was also studied in an angiogenesis bioassay since vessel growth is essential for ovarian follicle function. Immunohistochemistry revealed intense positivity for nesfatin-1 in swine granulosa cells in all developmental stage follicles. The precursor protein nucleobindin-2 (NUCB-2) showed a higher expression in granulosa cells from large follicles compared to medium and small follicles. In addition, our results demonstrated that nesfatin-1 stimulates cell proliferation and progesterone production and interferes with redox status by modifying NO production and non enzymatic scavenging activity in granulosa cells from large follicles. Moreover, nesfatin-1 displays a stimulatory effects on angiogenesis. Present study demonstrates, for the first time, that nesfatin-1 is physiologically present in the swine ovarian follicle where it could impair granulosa cell functions