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Introduction: Idiopathic pulmonary fibrosis (IPF) is a progressive, fatal lung disease with a poor prognosis and increasing incidence. Pirfenidone and nintedanib are the only approved treatments for IPF but have limited efficacy and their mechanisms of action are poorly understood. Here we have examined the effects of pirfenidone and nintedanib in a human model of lung fibrogenesis, and compared these with the putative anti-fibrotic compounds Lipoxin A4 (LXA4), and senicapoc, a KCa3.1 ion channel blocker. Methods: Early fibrosis was induced in cultured human lung parenchyma using TGFß1 for 7 days, ± pirfenidone, nintedanib, or LXA4. Pro-fibrotic responses were examined by RT-PCR, immunohistochemistry and soluble collagen secretion. Results: Thirty six out of eighty four IPF and fibrosis-associated genes tested were significantly upregulated by TGFß1 in human lung parenchyma with a ≥0.5 log2FC (n = 32). Nintedanib (n = 13) reduced the mRNA expression of 14 fibrosis-associated genes including MMPs (MMP1,-4,-13,-14), integrin α2, CXCR4 and PDGFB, but upregulated α-smooth muscle actin (αSMA). Pirfenidone only reduced mRNA expression for MMP3 and -13. Senicapoc (n = 11) previously attenuated the expression of 28 fibrosis-associated genes, including αSMA, several growth factors, collagen type III, and αV/ß6 integrins. Pirfenidone and nintedanib significantly inhibited TGFß1-induced fibroblast proliferation within the tissue, but unlike senicapoc, neither pirfenidone nor nintedanib prevented increases in tissue αSMA expression. LXA4 was ineffective. Conclusions: Pirfenidone and nintedanib demonstrate modest anti-fibrotic effects and provide a benchmark for anti-fibrotic activity of new drugs in human lung tissue. Based on these data, we predict that the KCa3.1 blocker senicapoc will show greater benefit than either of these licensed drugs in IPF.
RESUMO
OBJECTIVE: To compare flow cytometric data (ploidy and proliferative activity or percentage SG2M-phase cells) to cytologic and histologic data of the bladder carcinomas. MATERIALS AND METHODS: Cytologic and flow cytometric analysis of DNA content were performed on 48 bladder washings: 28 bladder washings from patients being followed for urothelial carcinomas and 20 control washings from individuals undergoing cytoscopy for other reasons. RESULTS: Cytological sensitivity and specificity of bladder washing were 75% and 91% respectively. Specificity was increased to 94% using flow cytometric DNA analysis whereas sensibility was moderately decreased to 68%. Combination of flow cytometry and cytology increased the diagnostic yield to 100%. The study of the patient group showed an increased abnormalities (aneuploidy and/or proliferate activity SG2M > 10%) according to the tumor grading and tumor staging. A cytometric test was positive in 80% for G3 tumours and in 68% for G2 tumours. The staging tumor was positive in 46%, 89% and 100% of the pTa-pT1, pT2 and pT4 tumours respectively. Otherwise the comparison of control group with patients showed a statistical correlation between cytometric test, staging tumour and tumoral grading as showed in the following groups: control/G1-G2 (p < 0.05), control/G3 (p < 0.001), control/pTa-pT (p < 0.05), control/pT2-pT4 (p < 0.001). CONCLUSION: We confirmed through this study the interest of the flow cytometric DNA analysis in the diagnosis and prognosis of bladder carcinomas, and we showed the importance of the histogram classification in order to facilitate their interpretation and to avoid the trap of false aneuploidy.
