曲尼司特对病毒性心肌炎小鼠心肌纤维化的作用

黄林枫; 文纯; 谢圭; 陈淳媛

doi:10.7499/j.issn.1008-8830.2014.11.017

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中国当代儿科杂志 ›› 2014, Vol. 16 ›› Issue (11) : 1154-1161. DOI: 10.7499/j.issn.1008-8830.2014.11.017

论著·实验研究

曲尼司特对病毒性心肌炎小鼠心肌纤维化的作用

黄林枫, 文纯, 谢圭, 陈淳媛

作者信息 +

Effect of tranilast on myocardial fibrosis in mice with viral myocarditis

HUANG Lin-Feng, WEN Chun, XIE Gui, CHEN Chun-Yuan

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文章历史 +

摘要

目的探讨曲尼司特在病毒性心肌炎心肌纤维化中的作用.方法 72 只BALB/C 小鼠随机分为对照组、模型组和干预组(n=24).模型组和干预组小鼠感染柯萨奇病毒B3 建立病毒性心肌炎模型,其中干预组小鼠造模后即用曲尼司特(200 mg/kg)灌胃给药直到取材前一天;对照组和模型组小鼠予等量生理盐水灌胃.造模后第7、14、28 天取各组8 只小鼠心肌组织,甲苯胺蓝染色、硫堇染色检测心脏肥大细胞数目,苏木精-伊红染色观察心肌组织病理学变化,Masson 三色染色观察心肌纤维化,RT-PCR、免疫组化检测骨桥蛋白(OPN)、转化生长因子-β1(TGF-β1)的mRNA 及蛋白表达,并对OPN mRNA 与肥大细胞数目作相关性分析.结果模型组和干预组小鼠心肌组织OPN mRNA 及蛋白的表达于第7 天达最高,第14 天次之,第28 天最低;TGF-β1 mRNA 及蛋白的表达第7 天升高,第14 天达高峰,第28 天较前稍下降;干预组小鼠心肌组织TGF-β1、OPN mRNA 及蛋白在各时间点的表达均低于模型组(均P<0.05),但仍高于对照组(均P<0.05).肥大细胞的数目与OPN 的表达呈正相关.结论曲尼司特可能通过减少肥大细胞数,抑制TGF-β1 及OPN 表达,在抗心肌纤维化过程中起重要作用.

Abstract

Objective To study the role of tranilast in the pathogenesis of myocardiac fibrosis in viral myocarditis.Methods Seventy-two BALB/C mice were randomly divided into control, model and intervention groups (n=24 each). Mice in the model and intervention groups were infected with Coxsackievirus B3 to induce viral myocarditis.The intervention group was given with tranilast (200 mg/kg) by gavage until sacrifice for sampling, while the other twogroups were administered with the same volume of normal saline. Cardiac tissues were obtained from 8 mice on 7, 14and 28 days after modeling. The mast cell number was observed by toluidine blue staining and thionine staining. Thecardiac tissues were stained with hematoxylin and eosin as well as masson trichrome to observe the pathological changesin cardiac tissues. The mRNA and protein expression of osteopontin and transforming growth factor-β1 was measuredby RT-PCR and immunohistochemistry respectively. Results In the model group, the mRNA and protein expression ofosteopontin reached the highest level on the 7th day, decreasing from the 14th day, and became to the least on the 28thday; while the expression of TGF-β1 increased from the 7th day, reaching a peak on the 14th day, and decreased slightlyon the 28th day. The mRNA and protein expression of TGF-β1 and OPN was lower in the intervention group thanthe model group (P<0.05), but higher than the control group (P<0.05). The expression of OPN mRNA was positivelycorrelated to the number of mast cells. Conclusions Tranilast can reduce myocardial fibrosis by decreasing the numberof mast cells, inhibiting the expression of TGF-β1 and OPN.

导出引用

黄林枫, 文纯, 谢圭, 陈淳媛. 曲尼司特对病毒性心肌炎小鼠心肌纤维化的作用[J]. 中国当代儿科杂志. 2014, 16(11): 1154-1161 https://doi.org/10.7499/j.issn.1008-8830.2014.11.017

HUANG Lin-Feng, WEN Chun, XIE Gui, CHEN Chun-Yuan. Effect of tranilast on myocardial fibrosis in mice with viral myocarditis[J]. Chinese Journal of Contemporary Pediatrics. 2014, 16(11): 1154-1161 https://doi.org/10.7499/j.issn.1008-8830.2014.11.017

参考文献

[1] Hua W, Jiang JB, Rong X, et al. The dual role of the cystathionine γ-lyase/hydrogen sulfide pathway in CVB3-induced myocarditis in mice[J]. Biochem Biophys Res Commun, 2009, 388(3):595-600.
[2] Said E, Said SA, Elkashef WF, et al. Tranilast ameliorates impaired hepatic functions in Schistosoma mansoni-infected mice[J]. Inflammophar, 2012, 20(2):77-87.
[3] Tan SM, Zhang Y, Cox AJ, et al. Tranilast attenuates the upregulation of thioredoxin-interacting protein and oxidative stress in an experimental model of diabetic nephropathy[J]. Nephrol Dial Transplant, 2011, 26(1):100-110.
[4] See F, Watanabe M, Kompa AR, et al. Early and delayed tranilast Treatment reduces pathological fibrosis following myocardial infarction[J]. Heart Lung Circ, 2013, 22(2):122-132.
[5] Fairweather D, Rose NR. Coxsackievirus-induced myocarditis in mice:a model of autoimmune disease for studying immunotoxicity[J]. Methods, 2007, 41(1):118-122.
[6] Rezkalla S, Kloner RA, Khatib G, et al. Effect of metoprolol on the acute phase of coxsackievirus B3 murine myocarditis[J]. J Am Coll Cardiol, 1988, 12(2):412-414.
[7] Azuma H, K.Banno T, Yoshimura, et al. Pharmacological properties of N-(3', 4'-dimethoxycinnamoyl) anthranilic acid (N-5'), a new anti-atopic agent[J]. Br J Pharmacol, 1976, 58(4):483-488.
[8] 李辉, 黄林枫, 文纯, 等. 心脏肥大细胞及TLR4 在小鼠病毒性心肌炎的作用[J]. 中国当代儿科杂志, 2013, 15(10):896-902.
[9] Engels W, Reiters PH, Daemen MJ, et al. Transmural changes in mast cell density in rat heart after infarct induction in vivo[J]. J Pathol, 1995, 177(4):423-429.
[10] Krishnamurthy P, Peterson JT, SubramanianV, et al. Inhibition of matrix metalloproteinases improves left ventricular function in mice lacking osteopontin after myocardial infarction[J]. Mol Cell Biochem, 2009, 322(1-2):53-62.
[11] 孙跃女, 陈淳媛, 杨作成, 等. 柯萨奇病毒B3 诱导体外培养心肌成纤维细胞增殖与骨桥蛋白表达[J]. 中华儿科杂志, 2008, 46(12):930-931.
[12] 蔡姿丽, 杨敏, 黄林枫, 等. 病毒性心肌炎小鼠心肌骨桥蛋白和Ⅰ型胶原表达的动态变化[J]. 中南大学学报医学版, 2012, 37(3):271-277.
[13] 成建定, 陈玉川, 胡丙杰, 等. 病毒性心肌炎急性期心肌胶原增生与TGF-β1 的表达[J]. 法医学杂志, 2000, 16(4):208-216.
[14] 聂宏刚, 关振中, 侯桂英, 等. 反复CVB3 感染对小鼠心肌的影响[J]. 中国地方病学杂志, 2003, 22(4):295-297.
[15] 韩福生, 孙辉, 刘立志, 等. TIMP-1 mRNA 表达在病毒性心肌炎小鼠心肌胶原重构中的作用及其与TGF-β1 的关系[J]. 中国地方病学杂志, 2005, 24(6):604-606.
[16] Pinto YM, Pinto-Sietsma SJ, Philipp T, et al. Reduction in left ventricular messenger RNA for transforming growth fact or beta(1) attenuates left ventricular fibrosis and improves survival without lowering blood pressure in the hypertensive TGR(mRen2) 27 rat[J]. Hypertens, 2000, 36(5):747-754.
[17] Hocher B, Godes M, Oliver J, et al. Inhibition of left ventricular fibrosis by tranilast in rats with renovascular hypertension[J]. J Hypertens, 2002, 20(4):745-751.
[18] Schipper ME, Scheenstra MR, van Kuik J, et al. Osteopontin:a potential biomarker for heart failure and reverse remodeling after left ventricular assist device support[J]. J Heart Lung Transplant, 2011, 30(7):805-810.
[19] Nagasaka A, Matsue H, Matsushima H, et al. Osteopontin is produced by mast cells and affects IgE-mediated degranulation and migration of mast cells[J]. Eur J Immunol, 2008, 38(2):489-499.
[20] Bulfone-Paus S, Paus R. Osteopontin as a new player in mast cell biology[J]. Eur J Immunol, 2008, 38(2):338-341.
[21] Ström Å, Franzén A, Wängnerud C, et al. Altered vascular remodeling in osteopontin-deficient atherosclerotic mice[J]. J Vasc Res, 2004, 41(4):314-322.
[22] Trueblood NA, Xie Z, Communal C, et al. Exaggerated left ventricular dilation and reduced collagen deposition after myocardial infarction in mice lacking osteopontin[J]. Circ Res, 2001, 88(10):1080-1087.