STZ诱导糖尿病联合正常饮食促ApoE-/-小鼠动脉粥样硬化的作用

doi:10.6040/j.issn.1671-7554.0.2014.273

摘要/Abstract

摘要： 目的探讨链脲佐菌素（STZ）诱导糖尿病联合正常饮食对ApoE-/-小鼠动脉粥样硬化病变的作用。方法取60只8周龄雄性ApoE-/-小鼠正常饮食饲养2周后，随机分为糖尿病组（采用小剂量STZ持续腹腔注射5 d）和非糖尿病对照组（柠檬酸盐缓冲液注射5 d），每组30只，正常饮食喂养13周后处死。分别检测小鼠血脂、炎症因子和血糖变化；HE和油红O染色检测各组小鼠大体和主动脉根部斑块病变；免疫组化、RT-PCR和Western blotting技术分别检测斑块内血管平滑肌细胞、巨噬细胞、基质金属蛋白酶-9/基质金属蛋白酶抑制剂-1（MMP-9/TIMP-1）、肿瘤坏死因子（TNF-α）和巨噬细胞趋化性因子（MCP-1）基因或蛋白含量变化。结果非糖尿病对照组主动脉及其根部病变轻微。与非糖尿病对照组比较，糖尿病组斑块面积明显增加，其中脂质、巨噬细胞、Ⅰ和Ⅲ型胶原、血管壁平滑肌细胞含量增加，但纤维帽处平滑肌细胞减少，斑块易损指数增加；血清MIF和IL-6水平升高，血管组织TNF-α、MCP-1、MMP-2和MMP-9基因表达或蛋白含量增加，而TIMP-1含量下降。结论 STZ诱导糖尿病ApoE-/-小鼠联合正常饮食有促动脉粥样硬化作用，并增加斑块易损性和机体炎症状态。

关键词: 链脲佐菌素, 动脉粥样硬化, ApoE-/-小鼠, 糖尿病

Abstract: Objective To explore the effects of streptozotocin(STZ)-induced diabetes combined with chow diet on atherosclerotic lesions of apoE-/-mice. Methods After 2 weeks of chow diet, 60 male apoE-/-mice (10 weeks old) were randomly divided into two groups. Diabetic apoE-/-mice (n=30) were constructed by intraperitoneal injection of low-dose STZ for 5 consecutive days. ApoE-/-mice (n=30) injected with citrate buffer liquid served as non-diabetic controls. All mice were fed with chow diet until sacrifice (23 weeks old). Serum lipid, inflammation cytokines and blood glucose were measured. Atherosclerotic plaques in the aorta and aortic root were examined with HE staining and Oil-Red-O staining. The mRNA and protein levels of vascular smooth muscle cells (SMCs), macrophages, MMP-9/TIMP-1, TNF-α and MCP-1 in the plaques were detected with immunohistochemical staining, RT-PCR and Western blotting, respectively. Results The non-diabetic apoE-/-mice showed slight atherosclerotic lesions, while diabetic apoE-/-mice had significantly aggravated atherosclerotic lesions. Compared with non-diabetic mice, diabetic apoE-/-mice had markedly larger size of plaques, in which lipid, macrophages, type Ⅰ and Ⅲ collagens and SMCs increased,while SMCs in fiber cap decreased, and atherosclerotic plaque instability index rose. Moreover, serum IL-6 and MIF levels, TNF-α, MCP-1, MMP-2 and MMP-9 mRNA all increased while TIMP-1 decreased. Conclusion STZ-induced diabetic apoE-/-mice fed with chow diet show deteriorated atherosclerotic lesions and relatively unstable plaques.

Key words: Diabetes mellitus, Atherosclerosis, ApoE-/-mice, Streptozotocin

中图分类号:

R541.4

孙慧, 赵磊, 甄茜, 王莎莎, 黄山英, 贺红, 胡琴. STZ诱导糖尿病联合正常饮食促ApoE-/-小鼠动脉粥样硬化的作用[J]. 山东大学学报（医学版）, 2014, 52(10): 1-8,28.

SUN Hui, ZHAO Lei, ZHEN Xi, WANG Shasha, HUANG Shanying, HE Hong, HU Qin. Pro-atherosclerotic effects of streptozotocin-induced diabetes combined with chow diet on apoE-/-mice[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(10): 1-8,28.

参考文献

[1] Juutilainen A, Lehto S, Ronnemaa T, et al. Similarity of the impact of type 1 and type 2 diabetes on cardiovascular mortality in middle-aged subjects[J]. Diabetes Care, 2008, 31(4): 714-719.
[2] Grundy S M, Howard B, Smith S Jr, et al. Prevention conference VI: Diabetes and cardiovascular disease executive summary conference proceeding for healthcare professionals from a special writing group of the American heart association[J]. Circulation, 2002, 105(18): 2231-2239.
[3] Toso C, Emamaullee J A, Merani S, et al. The role of macrophage migration inhibitory factor on glucose metabolism and diabetes[J]. Diabetologia, 2008, 51(11): 1937-1946.
[4] Tse J, Martin-McNaulty B, Halks-Miller M, et al. Accelerated atherosclerosis and premature calcified cartilaginous metaplasia in the aorta of diabetic male Apo E knockout mice can be prevented by chronic treatment with 17 beta-estradiol[J]. Atherosclerosis, 1999, 144(2): 303-313.
[5] Jawień J, Nastałek P, Korbut R. Mouse models of experimental atherosclerosis[J]. J Physiol Pharmacol, 2004, 55(3): 503-517.
[6] Hsueh W, Abel E D, Breslow J L, et al. Recipes for creating animal models of diabetic cardiovascular disease[J]. Circ Res, 2007, 100(10): 1415-1427.
[7] Nakashima Y, Plump A S, Raines E W, et al. ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree[J]. Arterioscler Thromb, 1994, 14(1): 133-140.
[8] Abel E D, Kaulbach H C, Tian R, et al. Cardiac hypertrophy with preserved contractile function after selective deletion of GLUT4 from the heart[J]. J Clin Invest, 1999, 104(12): 1703-1714.
[9] Gallagher E J, Sun H, Kornhauser C, et al. The effect of dipeptidyl peptidase-IV inhibition on bone in a mouse model of type 2 diabetes[J]. Diabetes Metab Res Rev, 2014, 30(3): 191-200.
[10] Naveiras O, Nardi V, Wenzel P L, et al. Bone-marrow adipocytes as negative regulators of the haematopoietic microenvironment[J]. Nature, 2009, 460(7252): 259-264.
[11] Kadoglou N P, Moustardas P, Kapelouzou A, et al. The anti-inflammatory effects of exercise training promote atherosclerotic plaque stabilization in apolipoprotein E knockout mice with diabetic atherosclerosis[J]. Eur J Histochem, 2013, 57(1): 3.
[12] Goldberg I J, Dansky H M. Diabetic vascular disease: an experimental objective[J]. Arterioscler Thromb Vasc Biol, 2006, 26(8): 1693-701.
[13] Tong C, Morrison A, Yan X, et al. Macrophage migration inhibitory factor deficiency augments cardiac dysfunction in Type 1 diabetic murine cardiomyocytes[J]. J Diabetes, 2010, 2(4): 267-274.
[14] Kunjathoor V V, Wilson D L, LeBoeuf R C. Increased atherosclerosis in streptozotocin-induced diabetic mice[J]. J Clin Invest, 1996, 97(7): 1767-1773.
[15] Candido R, Jandeleit-Dahm K A, Cao Z, et al. Prevention of Accelerated Atherosclerosis by Angiotensin-Converting Enzyme Inhibition in Diabetic Apolipoprotein E-Deficient Mice[J]. Circulation, 2002, 106(2): 246-253.
[16] Koïtka A, Cao Z, Koh P, et al. Angiotensin Ⅱ subtype 2 receptor blockade and deficiency attenuate the development of atherosclerosis in an apolipoprotein E-deficient mouse model of diabetes[J]. Diabetologia, 2010, 53(3): 584-592.
[17] Shen X, Bornfeldt K E. Mouse models for studies of cardiovascular complications of type 1 diabetes[J]. Ann N Y Acad Sci, 2007, 1103: 202-217.
[18] Libby P. Inflammation in atherosclerosis[J]. Nature, 2002, 420(6917): 868-874.
[19] Göran K, Hansson P L, Yan Usa Z. Innate and Adaptive Immunity in the Pathogenesis of Atherosclerosis[J]. Circulation Research, 2002, 91(4): 281-291.
[20] van der Wal A C, Becker A E, van der Loos C M, et al. Site of intimal rupture or erosion of thrombosed coronary atherosclerotic plaques is characterized by an inflammatory process irrespective of the dominant plaque morphology[J]. Circulation, 1994, 89(1): 36-44.
[21] Boring L, Gosling J, Cleary M, et al. Decreased lesion formation in CCR2-/-mice reveals a role for chemokines in the initiation of atherosclerosis[J]. Nature, 1998, 394(6696): 894-897.
[22] Huber S A, Sakkinen P, Conze D, et al.Interleukin-6 exacerbates early atherosclerosis in mice[J]. Arterioscler Thromb Vasc Biol, 1999, 19(10): 2364-2367.
[23] Wong N D, Patao C, Malik S, et al. Preventable coronaryheart disease events from control of cardiovascular risk factors in US adults with diabetes (Projections from Utilizing the UKPDS Risk Engine)[J]. Am J Cardiol, 2014, 113(8): 1356-1361.
[24] Sherry C L, O'Connor J C, Kramer J M, et al. Augmented lipopolysaccharide-induced TNF-alpha production by peritoneal macrophages in type 2 diabetic mice is dependent on elevated glucose and requires p38 MAPK[J]. J Immunol, 2007, 178(2): 663-670.
[25] Gough P J, Gomez I G, Wille P T, et al. Macrophage expression of active MMP-9 induces acute plaque disruption in apoE-deficient mice[J]. J Clin Invest, 2006, 116(1): 59-69.
[26] Ding S, Zhang M, Zhao Y, et al. The role of carotid plaque vulnerability and inflammation in the pathogenesis of acute ischemic stroke[J]. Am J Med Sci, 2008, 336(1): 27-31.
[27] Matsumura S, Iwanaga S, Mochizuki S, et al. Targeted deletion or pharmacological inhibition of MMP-2 prevents cardiac rupture after myocardial infarction in mice[J]. J Clin Invest, 2005, 115(3): 599-609.
[28] Newby A C. Matrix metalloproteinases regulate migration, proliferation, and death of vascular smooth muscle cells by degrading matrix and non-matrix substrates[J]. Cardiovasc Res, 2006, 69(3): 614-624.
[29] Allaire E, Forough R, Clowes M, et al. Local overexpression of TIMP-1 prevents aortic aneurysm degeneration and rupture in a rat model[J]. J Clin Invest, 1998, 102(7): 1413-1420.
[30] Rouis M, Adamy C, Duverger N, et al. Adenovirus-mediated overexpression of tissue inhibitor of metalloproteinase-1 reduces atherosclerotic lesions in apolipoprotein E–deficient mice[J]. Circulation, 1999, 100(5): 533-540.
[31] Lemaître V, Soloway P D, D'Armiento. Increased medial degradation with pseudo-aneurysm formation in apolipoprotein E-knockout mice deficient in tissue inhibitor of metalloproteinases-1[J]. Circulation, 2003, 107(2): 333-338.

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