The Influnce Of HDAC6 Inhibitor On Inflammation Cytokine Expression In Cells And Clinical Analysis Of COPD Accompanying Pneumonia

Objective: To discuss the inhibition of inflammation cytokine in diseased human brochial endothelial cells and human pulmonary arterial endothelial cells by selective HDAC6 inhibitor. Endotoxemia was induced by intraperitoneal injection of 7.5mg/kg LPS.Lung tissue was collected to assay MPO production and inflammation cytokine expression.To discuss the clinical sence of selective HDAC6 inhibitor by analysising information of 76 patients with COPD accompanying pneumonia.Method: Part1: DHBEs were pre-treated with Tubastatin A 10μM for 6h, then challenged with TNFα 20 ng/ml for 6 h. Extract the whole cell protein. Use BCA kit to assay protein concertration. TNF-α-induced IL-33 expression was measured in DHBEs by Western blot. Part2:HPAECs were pre-treated with Tubastatin A 10μM for 6h, then challenged with TNFα 20 ng/ml for 6 h.Extract the whole cell protein. Use BCA kit to assay protein concertration.TNF-α-induced VCAM-1 expression was measured in HPAECs by Western blot. Then discuss the cell signal. Part3:Endotoxemia was induced by intraperitoneal injection of 7.5 mg/kg LPS in phosphate-bufferedsaline(PBS).Tubastatin A 30mg/kg pretreat for 6h. Experiments were terminated 24 h after LPS challenge. Lung tissues were collected and homogenized in lysisbuffer.Use BCA kit to assay protein concertration. VCAM-1 expression was measured by Western blot. Part4:Clinical analysis of 76 patients with COPD accompanying pneumonia from January 2015 to March 2016.Result: 1. Tubastatin A can reduce IL-33 expression in DHBEc. 2. Tubastatin A can reduce VCAM-1 expression in HPAEc.3.Tubastatin A inhibits TNF-α-activated Jak3/STAT-1 pathways which were involved in the TNF-α-induced VCAM-1 expression. 4.Tubastatin A inhibits TNF-α-activated Jak3/STAT-1 pathways through tyrosine phosphorylation. 5.Tubastatin A does not inhibit the expression of p-STAT1(Ser). 6.IRF-1, which is necessary for TNF-α-stimulated VCAM-1, is regulated by Tubastatin A. 7.Tubastatin A inhibits MPO production in sepsis model. 8.Tubastatin A inhibits VCAM-1 and IRF-1expression in sepsis model. 9.The effective rate of 76 patients with COPD accompanying pneumonia is 77.63% after treatment. 10.There are 34 bacterial strains by bacterial culture from phlegm of 76 patients with COPD accompanying pneumonia. Most of them are candida albicans(47.06%).Conclusion: 1.HDAC6 inhibitor can reduce TNF-α-induced IL-33 expression in DHBEc. 2. HDAC6 inhibitor can reduce TNF-α-induced VCAM-1 expression in HPAEc,which through inhibits TNF-α-activated Jak3/STAT-1 pathways of tyrosine phosphorylation instead of serine. 3. HDAC6 inhibitor can reduce lung tissue MPO production and VCAM-1 expression in LPS-induced sepsis model. 4. The effective rate of 76 patients with COPD accompanying pneumonia is 77.63% after treatment. There are 34 bacterial strains by bacterial culture from phlegm of 76 patients with COPD accompanying pneumonia. Most of them are candida albicans(47.06%).