1、 u我国糖尿病患病率我国糖尿病患病率9.7%9.7%， 其中其中2 2型糖尿病（型糖尿病（T2DM)T2DM)占占90%90%，绝，绝大多数大多数T2DMT2DM患者存在患者存在IRIR；uIRIR定义定义：是指胰岛素效应器官对正常剂量胰岛素反应性降低，是指胰岛素效应器官对正常剂量胰岛素反应性降低， 导致胰岛素敏感组织（脂肪、肝脏和肌肉）对导致胰岛素敏感组织（脂肪、肝脏和肌肉）对 葡萄糖摄取和利用障碍。葡萄糖摄取和利用障碍。 胰岛素抵抗（胰岛素抵抗（Insulin resistance, IR)Insulin resistance, IR)Reaven,1988,DiabetesReaven,
2、1988,Diabetes胰岛素（胰岛素（Insulin)Insulin)的生理功能的生理功能Saltiel AR, Nature, 2001 n 胰岛胰岛 细胞分泌的促合成激素细胞分泌的促合成激素;n 促进碳水化合物、脂类和蛋白质合成：刺激葡萄糖、氨基促进碳水化合物、脂类和蛋白质合成：刺激葡萄糖、氨基酸和脂肪酸转运入细胞，提高糖原、脂类和蛋白质合成相酸和脂肪酸转运入细胞，提高糖原、脂类和蛋白质合成相关酶的表达和活性；关酶的表达和活性；n 抑制碳水化合物、脂类和蛋白质降解相关酶类的表达，抑抑制碳水化合物、脂类和蛋白质降解相关酶类的表达，抑制其分解及释放入血。制其分解及释放入血。胰岛素抵抗综合征
3、（胰岛素抵抗综合征（IRS)IRS) 的特征的特征肥胖肥胖 （尤腹型肥胖（尤腹型肥胖 visceralvisceral obesityobesity）糖耐量减低糖耐量减低 (Glucose(Glucose intorlerance:intorlerance: impairedimpaired glucoseglucose intolerance;intolerance; impairedimpaired fastingfasting glucose,glucose, T2DM)T2DM) 脂代谢紊乱脂代谢紊乱（highhigh triacylglycerol,triacylglycerol,
4、lowlow HDL,HDL, small dense LDL small dense LDL particles)particles)高血压高血压内皮功能紊乱内皮功能紊乱动脉粥样硬化性冠心病动脉粥样硬化性冠心病 (Atherosclerotic(Atherosclerotic CVD)CVD)高胰岛素血症高胰岛素血症胰岛素抵抗胰岛素抵抗Defronzo RA et al, 2010,Diabetologia 生理及生理及IRIR时的胰岛素受体后信号通路时的胰岛素受体后信号通路IRS：胰岛素受体底物胰岛素受体底物（Insulin receptor substrate）PI3K: 磷脂酰肌醇磷脂
5、酰肌醇-3-激酶激酶GLUT-4：葡萄糖转运体葡萄糖转运体 4 (Glucose transporters) HealthyIRDefronzo RA et al, 2010,DiabetologiaHypertensionAthersclerosis u胰岛素抵抗人群胰岛素抵抗人群CVDCVD发病率显著升高发病率显著升高u可能机制：可能机制：1.1.代偿性高胰岛素血症代偿性高胰岛素血症抑制肝脏产生和释放过多葡萄糖，但内抑制肝脏产生和释放过多葡萄糖，但内脏脂肪有过多脂肪分解导致游离脂肪酸释放，并抑制肌肉摄脏脂肪有过多脂肪分解导致游离脂肪酸释放，并抑制肌肉摄取和利用葡萄糖；取和利用葡萄糖；FFA
6、FFA通过脂毒性促进通过脂毒性促进ASAS的发生的发生2.2.胰岛素抵抗伴随的高血压、高血糖、高血脂等因素导致胰岛素抵抗伴随的高血压、高血糖、高血脂等因素导致内皮内皮功能失调功能失调为起始环节的为起始环节的ASAS的发生。的发生。3.3.颈动脉内膜、中膜增厚颈动脉内膜、中膜增厚是糖尿病患者亚临床是糖尿病患者亚临床ASAS的标志，胰岛的标志，胰岛素敏感性与颈总动脉内膜、中膜厚度呈负相关。素敏感性与颈总动脉内膜、中膜厚度呈负相关。 胰岛素抵抗与动脉粥样硬化性冠心病相关胰岛素抵抗与动脉粥样硬化性冠心病相关Defronzo RA et al, 2010,Diabetologia 肥胖肥胖 (Obesi
7、ty(Obesity），），IRIR 和和T2DMT2DM 肥胖尤其是腹型肥胖，是引起IR和T2DM最重要的危险因素。 循证医学研究证据：循证医学研究证据：1.美国肥胖青少年中IR发生率约50%；2.临床T2DM病例约80% 超重或肥胖；减重、减少内脏脂肪体积等措施可切实改善IR3.即使体脂肪分布“正常”，脂肪功能失调脂肪功能失调（adipose tissue dysfunction） 与IR及T2DM的发生发展密切相关。Chan JM,1994, S.Matthaei et al, 2000, Goossens GH,2008腹部脂肪堆积腹部脂肪堆积（Visceral Fat Deposit
8、ion）Foster MT, Adipocyte,2012Visceral ( 10% of body total，VAT), is highly associated with obesity-related health consequences中心型肥胖中心型肥胖 （Central Obesity）与与IRIR Foster MT, Adipocyte,2012VazquezVela M E, 2008Planat-Benard V,2004Masoodi M, 2014uPluripotential fibroblasts myocytes, chondrocytes and adip
9、ocytesCD34, CD13 positive TZDs: ( 噻唑烷二酮类噻唑烷二酮类) PPAR agonist 如罗格列酮，匹格列酮如罗格列酮，匹格列酮 PPAR: Peroxisome proliferators -activated receptor 过氧化物酶增殖体激活的受体过氧化物酶增殖体激活的受体 PGC-1：peroxisome proliferator -activated receptor- coactivator-1 过氧化物酶体增殖物激活受体过氧化物酶体增殖物激活受体 辅激活因子辅激活因子 C/EBP：CCAAT增强子结合蛋白增强子结合蛋白Adipogenesis
10、,lipogenesis SREBP-1c: adipocyte determination and differentiation dependent factor 1 (ADD1) 脂肪细胞定向分化因子脂肪细胞定向分化因子1 PGC-1 脂肪生成（脂肪生成（AdipogenesisAdipogenesis）Rosen ED,2006;Masoodi M 2014 脂肪细胞分化的相关信号通路及其细胞内外信号调节脂肪细胞分化的相关信号通路及其细胞内外信号调节促成脂家族促成脂家族:KLFs, STATs1,5A 5B. 抑成脂家族抑成脂家族:GATA2,3; SIRT1Preadipocytes
11、 （前脂肪细胞）（前脂肪细胞）Small adipocytesLarge adipocytesPPAR: Peroxisome proliferators -activated receptor 过氧化物酶增殖体激活的受体过氧化物酶增殖体激活的受体PGC-1：peroxisome proliferator-activated receptor- coactivator-1 过氧化物酶体增殖物激活受体过氧化物酶体增殖物激活受体 辅激活因子辅激活因子TZDs: 噻唑烷二酮类，噻唑烷二酮类，PPAR agonist 如罗格列酮，匹格列酮如罗格列酮，匹格列酮 C/EBP-PPAR脂肪细胞容积调控与脂肪
12、细胞容积调控与IRIR发生密切相关发生密切相关Rosen ED,2006; Lundgren M, Diabetologia,2007增生：增生：Hyperplasia肥大：肥大：HypertrophyVazquezVela M E, 2008LPL： 脂蛋白脂酶脂蛋白脂酶ATGL: 脂肪三酯酰甘油酯酶脂肪三酯酰甘油酯酶HSL: 激素敏感酯酶激素敏感酯酶Peripilipin: 围脂滴蛋白围脂滴蛋白脂肪代谢：生脂作用（脂肪代谢：生脂作用（lipogenesis)lipogenesis) 脂解反应（脂解反应（lipolysis)lipolysis)乙酰辅酶乙酰辅酶A羧化酶羧化酶脂肪酸合成酶脂肪酸
13、合成酶ChREBPEnlargedadipocytesObesity: 肥胖肥胖Adipocyte: 脂肪细胞脂肪细胞Adipokines: 脂肪因子脂肪因子Modified from Goossens GH, 2008; Smith U, 2002(PMID:12080441)Adiponectin: 脂联素脂联素 Leptin: 瘦素瘦素 TNF- : 肿瘤坏死因子肿瘤坏死因子 IL-6: 白介素白介素6 Resistin: 抵抗素抵抗素脂肪功能失调：脂肪因子分泌紊乱和炎症脂肪功能失调：脂肪因子分泌紊乱和炎症 Lean stateIR or obesity stateCao H, 2014
14、Obesity, FFA, inflammation and IRSchenkS,2008;Skurk T, 2007,TLR4 signalingNF B signaling内质网应激是内质网应激是IRIR发生的重要机制发生的重要机制Beck and Kaufman, 2012未折叠反应：未折叠反应： Unfolded protein response Unfolded protein response （UPRUPR）内质网跨膜信号蛋白：内质网跨膜信号蛋白：PERKPERK，ATF6ATF6，IRE1IRE1 内质网应激与细胞凋亡内质网应激与细胞凋亡Carla J. H. van der
15、Kallen，2009，ApoptosisInflammationProliferation, differentiationPreadipocytes Small adipocytesSSensitivity to insulinproliferation, ,apoptosis differentiationThe percentage of small adipocytes, the size of adipocytesModified from Naoto Kubota, Molecular Cell 1999; Ishihara, Y，Atherosclerosis,2010; Le
16、e MH, Kidney Int,2008; Okuno A J Clin Invest ,1998, Skurk T, J Clin Endocrinol Metab 2007e.g.PravastatinPitovastatinSSSenstivity to insulinLLLThe plasma level of saturated free fatty acids (FFA)Adipocyte tissue dysfunctionEvidences:Duan D, et al. Nature. 1997;390(6658):417-421.Yamazaki J, et al. J P
17、hysiol. 1998;507 ( Pt 3):729-736.Wang GL, et al. Circ Res. 2002;91(10):E28-32.Zhou JG, et al. J Biol Chem. 2005;280(8):7301-7308.Zhang HN, et al. Apoptosis. 2006;11(3):327-336.Tang YB, et al. Cell Prolif. 2008;41(5):775-785.Mao J, et al. Biochem Pharmacol. 2008;75(9):1706-1716.Xu B, et al. Acta Bioc
18、him Biophys Sin (Shanghai).42(6):370-380.Zhu L, et al. Biochem Pharmacol.83(3):324-334.Guan YY, et al. Trends Pharmacol Sci. 2006;27(6):290-296.Sasaki S, et al. ClC family in the kidney, Jpn J Physiol. 1994;44 Suppl 2:S3-8.Liu J, et al. Apoptosis 2013; Qian Y, et al. Apoptosis 2011 Shi, XL, et al. H
19、ypertension 2007; Liu YJ，et al: Hypertension 2010 Sham Htn Htn+simv.1 W4 W8 W12 WThomas J. Jentsch, et al. The Journal of Neuroscience, October 15, 2008. 28(42):1058710598Serum insulin levels were significantly lower in ClC-3 KO miceClC-3 protein is important in the regulation of cellular apoptosisZ
20、hang HN, Apoptosis,2006; Qian Y, . Apoptosis 2011; Liu J, et al. Apoptosis 2013 ClC-3 encodes volume-regulated chloride channel in vascular smooth muscle cells and involved in VSMC proliferationWang GL, Cir. Res.2002, Zhou JG, JBC,2005,Guan YY, Trends Pharmacol.Sci.,2006Shi, XL, Hypertension 2007; L
21、iu YJ，Hypertension 2010 Sham Htn Htn+simv.1 W4 W8 W12 WStatinsClC-3Adipose tissue dysfunction during Insulin Resistance？Questions: 1, Whether there is a relation between ClC-3 and IR ? 2, If yes, what is the underlying mechanism? van Tienen FH, et al. Int J Obes (Lond).2011.35(9):1154-1164.Huang YY,
22、 20142 2型糖尿病大鼠肝脏及骨骼肌组织型糖尿病大鼠肝脏及骨骼肌组织ClC-3ClC-3蛋白质表达蛋白质表达与与IRIR无明显相关性无明显相关性 Huang YY, 2014脂肪脂肪ClC-3ClC-3蛋白表达与蛋白表达与TGTG、FFAFFA、磷酸化、磷酸化HSLHSL水平水平呈正相关呈正相关, ,与与adiponectinadiponectin水平呈负相关水平呈负相关 Huang YY, 2014Huang YY, 2014 Data are presented as means S.E., n=12. *P 0.05 vs WT group. *P 0.01 vs WT group.
23、 #P 0.05 vs WT-T2DM group. #P 0.01 vs WT-T2DM group.WTClC-3 KO WT-T2DMClC-3 KO-T2DMGlucose (mM) 5.740.40 6.320.8426.201.40*21.801.46#Insulin (IU/mL) 2.890.15 2.230.23* 3.130.102.610.27HOMA-IR 0.860.11 0.620.10 3.630.31* 2.380.19#AUC (ITT)13.321.2912.891.2936.355.92*23.444.33#Serum FFAs (mM) 0.650.05
24、 0.560.0350.880.08* 0.610.07#Serum TG (mM) 0.660.06 0.800.11 1.380.10* 0.930.17#Data are presented as means S.E., n=12.*P 0.05 vs WT group. *P 0.01 vs WT group. #P 0.05 vs WT-T2DM group. #P 0.01 vs WT-T2DM group.Effect of ClC-3 knockout on metabolic parameters of HSFD/STZ-treated diabetic miceHuang
25、YY, 2014Jentsch, TJ. J. Neurosci., 2008Serum insulin levels were significantly lower, but neither hyperglycemia nor rebound hypoglycemia in ClC-3 KO miceNeither hyperglycemia nor rebound hypoglycemia was shown following a glucose load in ClC-3-/- miceDickerson LW, Brain Res,2002.Huang YY, 2014Huang
26、YY, 2014Huang YY, 2014Huang YY, 2014ClC-3 siRNA significantly improved palmitate-induced apoptosis in 3T3-L1 preadipocytes Huang YY, 2014Huang YY, 2014 Huang YY, 2014van der Kallen, C. J. Apoptosis (2009)Guo,W, Am J Physiol Endocrinol Metab (2007)Back and Kaufman , Annu Rev Biochem(2012)eiF2ATF4CHOP
27、ER LumenGRP 78PERKprocaspase-12IRE1 TRAF2 ASK1caspase-9APOPTOSISJNKBcl-2Bak/Bax caspase-3caspase-12Huang YY, 2014ClC-3 siRNA alleviated palmitate-induced CHOP and ATF4 protein expression in preadipocytes Huang YY, 2014ClC-3 siRNA didnt affect palmitate-induced p-eiF2 or p-PERK protein expression in
28、preadipocytes ClC-3 siRNA alleviated palmitate-induced p-JNK1/2 protein expression in preadipocytes Huang YY, 2014ClC-3 siRNAeiF2ATF4CHOPER LumenGRP 78PERKIRE1 TRAF2 ASK1caspase-9APOPTOSISJNKBcl-2caspase-3PALMITATESummarySummary1.ClC-3 deficiency could improve the enhanced preadipocyte apoptosis, hy
29、perglycemia and hyperlipidmia, and insulin resistance during T2DM2. ClC-3 protein is required for ER-stress dependent preadipocyte apoptosis induced by palmitate, but not oleate. 3.These data suggest that ClC-3 may serve as a novel potential target to prevent metabolic disruption in the development of insulin resistance and T2DM
