ABHD2调控肺泡Ⅱ型上皮细胞凋亡 在慢性阻塞性肺疾病发生中的作用.ppt

Abhd2 regulates alveolar type apoptosis and airway smooth muscle remodeling: a key target of COPD research,Shoude Jin,Harbin Medical University, China,COPD - a silent killer,Insidious, diagnosed at late stages,Background,Pathophysiology,Normal vessel,Abormal vessel,Smoking is a major risk factor for COPD Only 10-20% of long-term heavy smokers develop into COPD Part COPD patients with no history of smoking,Environment,Genetics,Clinical characteristics and prevention direction of COPD :： Interaction of individual genetic susceptibility and environmental factors It is important of susceptible early screening and early intervention in the prevention of chronic obstructive pulmonary disease,The importance of genetic susceptibility to COPD,Recent Advances and trends: Genomic analyses to reveal genetic susceptibility to COPD Regional and Ethnic differences: 1-AT deficiency increase the risk of Caucasian COPD, it is a susceptibility gene for COPD TNF gene mutation associated with Asian COPD such as EPHX1, IL4, IL-10 associated with COPD In addition to 1-AT, it has not been determined that other genes are susceptibility gene for COPD,1,Our previous study IL-13 G + 2044A point mutations associated with Caucasian COPD, but no correlation with the Chinese COPD patients (2014, Molecular Biology Reports, Dr. Shoude Jin and colleagues),Key Findings: Abhd2 gene deletion in mice naturally develop into emphysema phenotype (2009, BBRC， Dr. Shoude Jin and colleagues) Abhd2 protein is a lipid hydrolase,ABHD2 homologous gene mutations can increase the risk of COPD （2015, Plos one， Dr. Shoude Jin and colleagues ）,2,3,Yamamura team (our collaborators) Kumamoto University, Japan,Abhd2 mouse gene expresses in alveolar type epithelial cells and airway smooth muscle cells,Trapping vector insertion point,Further to verify: Abhd2 expressed in alveolar type cells,Figure. -gal staining pattern P180 common immunofluorescence of 1.Abhd2 knockout mice lung tissue,Our research team: using the Abhd2 knockout mice,Table. the content and relatively of PC, DSPC and protein in lavage fluid and lung tissue homogenates,Phospholipid metabolism in lavage fluid,Increased apoptosis of alveolar type ,Abhd2Gt/Gt,Wild type,Figure . P180 antibody immunohistochemistry of 12-month-old mouse lung slices, the mouse type II alveolar cell count of 6 months and 12 months,The nnumbers of alveolar type cells in every 1000 lung cells,Increased macrophage infiltration in Lung tissue,The nnumbers of macrophages in every 1000 lung cells,12 months,Abhd2Gt/Gt wild type,Figure. F4 / 80 antibody immunohistochemistry and lung macrophages count comparison in Abhd2Gt / Gt and wild type lung tissue sections,6 months,Figure. The staining of elastic fiber,Wild type Abhd2Gt/Gt,Elastic fibrin reduction,Figure. the stained with HE of lung tissue,Alveolar destruction, expanding and spontaneously formed emphysema phenotype,Figure. Change of protease and anti-protease Factor,Protease /anti-protease imbalance,Figure. The expression of proinflammatory cytokines and apoptosis related genes at the mRNA level,Enhanced expression of inflammatory and apoptosis-related factors,Figure. The expression levels of oxidation and antioxidant factors,Oxidation and antioxidant factors indifference,Mouse Abhd2 playes an important role in emphysema,The exact mechanism was unclear,But the function of hABHD2 was unclear,Abhd2 features ： Abhd2 deficient mice spontaneously emphysema Abhd2 expressed in airway smooth muscle and vascular smooth muscle cells. Abhd2 deletion induced vascular smooth muscle cell migration and intimal hyperplasia,BT =bronchial tract,Clinical manifestations of COPD ： lung: airway remodeling, emphysema, vascular remodeling: smooth muscle cell proliferation, migration lung outside: peripheral muscle atrophy and dysfunction and other symptoms,We speculate: ABHD2 associated with COPD,interlobular vein,Protein Similarity 98.59%/425aa (from NCBI),mAbhd2 and hABHD2 Homology,Abhd2,Abhd family and Abhd2 features:,Alasdair J. Edgar and Julia M. Polak. Cloning and Tissue Distribution of Three Murine / Hydrolase Fold Protein cDNAs. Biochemical and Biophysical Research Communications 292, 617625 (2002),Origin: Edgar and colleagues cloned / hydrolase family encoding a protein from emphysema model mouse lung cDNA library in 2002 : / hydrolase gene 1 (Abhd1), Abhd2 and Abhd3 Structural domains: Abhd1, Abhd2 and Abhd3 may play an important role in gene function with the same structural domain,Study: Associations of ABHD2 Genetic variations with Risks for Chronic Obstructive Pulmonary Disease in a Chinese Han Population,Extraction and comparative analysis genomic DNA of COPD patients and normal population Screening gene SNP point of ABHD2 : from shared domain of Abhd2,The study found: Rs12442260 mutants (CT / CC ), upstream of the translation initiation site in the fifth ABHD2 gene intron from the sixth exon upstream 489bp, increased the risk of COPD Conclusion: ABHD2 gene polymorphism was associated with COPD risk,1,2,3,4,Exon 5,9,10,Exon 6,7,8,Abhd2 gene,Supplementary Table. Markers genotyped in the current study.,Outlook Analysis and screen specific mutations sites of ABHD2 in COPD patients Construct ABHD2 specific mutations knock humanized mouse by use of genetic manipulation techniques Determine causality of people ABHD2 specific locus mutation occurs with COPD Determine COPD susceptible early screening biomarkers,ABHD2 may be susceptibility gene of COPD But it is not clear that whether the specific site mutations affect gene function,科学问题,Abhd2 gene express in bronchial smooth muscle cells and vascular smooth muscle cells The study found: Abhd2 gene deletions may promote vascular smooth muscle cells Remodeling We hypothesized that: Abhd2 genes may be involved in airway smooth muscle remodeling ACOS,ACOS is the main reason for the late COPD patients died. COPD patients often appear ACOS phenotype with age and progression of the disease in COPD population over the age of 55 may be as high as 55%. Therefore, to effectively prevent the development of ACOS and treatment is essential for COPD,Our previous study: Abhd2 knockout mice was sensitized by ovalbumin to airway mucus hypersecretion and eosinophil Granulocyte infiltration, increased smooth muscle layer thickening, increased - myosin expression and other pathological features of asthma,Figure . Observate smooth muscle hyperplasia in Abhd2 knockout mice before and after the degree of airway by ovalbumin sensitized,Figure . Observate inflammatory cell infiltration of Abhd2 knockout mice before and after lung ovalbumin sensitized,ova sensitized,ova,ova sensitized,ova,Outlook: 1. Explore the possible relationship between Abhd2 gene polymorphism and asthma - COPD overlap syndrome-associated risk 2. Further to constructe humanized animal models of ABHD2 and to verify,We have constructed ACOS multivariate dangerous animal model We will further verify: Abhd2 may be involved in airway smooth muscle cells remodeling, and the possible relationship between the occurrence of ACOS,Thank You ！,