心力衰竭的信号通路课件幻灯

1 田小利田小利 人类群体遗传室人类群体遗传室 北京大学分子医学研究所北京大学分子医学研究所 死亡 代偿期失代偿期恶化期 结构和功能的改变-重塑 高血压，冠心病 遗传，心肌病 瓣膜病，先心病 * * 心力衰竭 心力衰竭的诱因和病理过程心力衰竭的诱因和病理过程 2 激素、药物、 神经递质 膜受体 第二信使 Ca2+ 离子通道 活性肽 心力 衰竭 心律 失常 线粒体活性氧 心脏病相关基因 活性肽 (1) (1) 心 力 衰 竭 心力衰竭可能的发展基础心力衰竭可能的发展基础 3 心力衰竭 临床诊断 遗传学 通道电生理学 神经和信号转导 心肌重塑的关键分子 细胞水平鉴定 动物模型 计算机数字化模拟 3-D结构研究 1. 作用机制及早期干预 2. 新药靶点及可药性 3. 早期诊断标记的可能性 目前主要研究的内容目前主要研究的内容 4 AOGEN Renin ACE ANG I ANG II 5 Salt/water balance Contraction of VSMC Mitogenic effects Neurotransmitter Hormone-stimulation factor Cardiac output 6 PLD PLC AII a g b PC PA DAG CholineIP3 PIP2 PKC Mek Mapk Stats ER Ca + Shc Jaks SIE C-fos C-Jun Stats SOS Grb2 Ras Raf GTP C-myc Egr Ca + Ca + Ca + Ca + 7 AOGEN Renin ACE ANG I ANG II Kallikrein Tonin Chymotrypsin Capthepsin G Chymase Kallikrein Chymotrypsin Capthesin G 8 9 50 40 30 20 10 0 Captopril Chymostatin Cap + Chymo Cap + Aprot CV 11974 %phenylephrine ANG1 perfusion isolated aorta Nishimura, 1998 Chymase: Ang II forming enzyme in vitro Found, cloned and characterized by Dr. Hidenori Urata Urata, 1990ACE Chymase Fe Ad Tian, 1996 Renin Angiotensinogen ACE Angiotensin I Angiotensin II AT1AT2 Angiotensinogen Renin ACE AT1 AT2 10 Angiotensinogen Renin Angiotensin 1 Angiotensin 2 Kininogen Kallikrein Bradykinin Inactive peptides ACE RAS (AII/AT1) AS (a1) ETS (ET-1/ETA) 11 12 Angiotensin 1 Bradykinin AcSDKP (N-acetyl-Ser-Asp-Lys-Pro) Angiotensin II Cell growth. etc. degraded fragments degraded fragments Actiate TGF- signaling 13 Fleming I et al. Physiology. 2005;20:91-5. ACE NH2 ACE inhibitor Extracellular Cytosol COOH Nucleus Gene expression (ACE, COX-2) CK2 JNK MKK7 P JNKcJun cJunP cJunPcJunP cJunP AP-1 cJunP cJunP Clinical significance of this pathway is under investigation Angiotensinogen (M235T) Together with ACE-D, associated male LVH CAD AT1 (A1166C) Associated male LVH 14 15 ACE (I/D) Earlier than 1994: tends to associated Later than 1994: tends to not associated LVH of man, not woman Degree, not frequency of LVH ACEI treatment QT dispersion, heart rate variability Exercise-induced LHV 16 Aorta Lung Kidney Brain Liver L. Ventricle R. ventricle Sk. muscle rACE hACE 17 SD TGR 50 60 70 80 90 100 110 -10-9-8,5-8-7,5 50 60 70 80 90 100 110 -10-9-8,5-8-7,5 SD TGR 30 25 20 15 10 5 0 SD TGR AII (pg/g) AI on coronary artery flowCardiac AII 18 ACEI 1,6 1,8 2,0 2,2 2,4 2,6 2,8 3,0 LV/B SD-sh SD-ab TGR-ab TGR-sh LV/B (mg/g) 0 1 2 3 4 5 6 7 8 ANFCol3 %GAPDH 0 20 40 60 80 100 120 140 160 180 SD-sh TGR-sh SD-ab TGR-ab SD-sh TGR-sh SD-ab TGR-ab ANF Col3 GAPDH SD-sh SD-ab TGR-ab TGR-sh SBP (mmHg) 19 20 Age 10 months col1 col3 GAPDH NTGR TGRL1172 TGRL1173 Lisinopril AcSDKP 21 ACE TGF- TGF-R1 Smad3Smad2 Smad4 Col TGF- GAPDH Smad3-p Smad3 Smad2-p Smad2 AcSDKP NTGR TGRL1172 TGRL1173 NTGR TGRL1172 TGRL1173 Lisinopril AcSDKP NTGR TGRL1172 TGRL1173 Lisinopril AcSDKP 22 23 Transcriptional machinery CmG H-act Epigenetic modification Definition of CG island in RAS promotersDefinition of CG island in RAS promoters 24 ATG TSS 5-UTR 1 kb CG sites CTGTCCCTGGCTCCTTCCCTGATCCCACCGCCAGCCTCACCCCACGGTTCCTCCATTGCC CCACCTCCCACTGCGCCGCCGGGCCTCTGCCAGGGTCAAGGGGCTTCCCCCCTCTGGCAG CAGACGCCATGGTGCCGAGGTGGCCTCCACAACCGCCCTGTGCGCCAATAGGACAAGACT GTCCTCCCTCCCCCACACTTGTCACTTTGAGGGACACGTGGATGAGACAGGAAAACACAG GGGAGTGTGGAGACCTGAGGTGACTTGGAGCAAGCCTCTCAACCTGAGCGGCAATTTCTT CATCTGTAAAATGAGGGGGTTGTTCTCATCTCTGAGGCTTTGTGTCGCTCTCAAAGCCTG CTAGCCTCGGGTTCTAGGACTCTGTTGGGATCGTGTGTGATGTTTTCTGCTGAGCGACGG CAGCCTGTGTCCTCGGGGGGAAAGAGGGCAGGCGCTCCAAAGCTCCTGCGCTCTGTGGCT CCCCCTCCCTCGCAGCCCCAAGCCCCAGGTGTGCCGGCCGCCCTGAGCCCCTCCAGCACC TCCCGGAGGCGCCTGCAAGACACCTAAGGTCCCCGCCTCCCTCCTCTCCCCCCCGCCACA CCCCTACCCCCGGCAGGCGACGTCCCCGCCCCTCGACCATGGCCTGGTGAAGAAGCCGGC CAGGCCCGATCAGCCCCATCCCCGCCGCACGAGCGGCGCCTGCGGACAGCTCCTGGGGCC CCGGCCTTGTCACTCCGGAGGCGGGAGGCTCCGGGGGGTCGGGCTGGGAAGATCGAGCCG GAGGCCGCTAGGCTCCCAGGCCCCGGCCGAGGCTGCGCGGCCGCACGGTGGGCAGGCTCG GGTGTTCCGGCAAACTGCCGGGTCCCCATCTTCAAAAGAGAGGAGGCCCTTTCTCCAGCT TCCTCTGCGGGAGCCCGACCCAGCCCCATCCCGCCACCCCCGGGCTGCACCTCGGCCCCT CCCCGGCCCGCGCCCCTGCCCGGGGCGGGCCAGGAACCTCGGCCCGCGCCGCTGGGGACT TTGGAGCGGAGGAGGAAGCGCGGCGGGGCGGGGGCGGGGGTGTGTCGGGTTTTATAACCC GCAGGGCGGCCGCGGCGCAGGAGAAGGGGCAGAGCCGAGCACCGCGCACCGCGTCATGGG ACEACE启动子区启动子区CGCG岛分析岛分析 25 26 Angiotensinogen Renin ACE AT1a AT2 74 69 87 45 88 甲基化分析的原理甲基化分析的原理 27 mCG 亚硫酸氢钠 mCG PCR CG CG UG TG CN UN TN 克隆测序 直接测序 Characteristics of ratsCharacteristics of rats SHR/SP （N=20）WKY（N=20）P value HR（bpm）376.834.6306.729.50.0000 SBP(mmHg)207.115.0130.610.30.0000 DBP(mmHg)164.814.995.211.90.0000 H/B(mg/g)4.160.013.690.0160.0000 V/B(mg/g)3.860.083.340.0170.0000 28 CCG 29 100% protected = 100% methylation How to quantify How to quantify methylationmethylation C WKYSHR 30 MethylationMethylation of AOG of AOG CGCC CGCC 31 GCCC GCCC WKYSHR MethylationMethylation of ACE of ACE (reversal sequence)(reversal sequence) 32 33 34