目的：探讨基质金属蛋白酶-9（MMP-9）在病毒性心肌炎（VMC）小鼠发病过程中的动态变化及意义。方法：以柯萨奇病毒 B3 感染 BALB/C 小鼠建立 VMC 模型，腹腔注射 0.1 mL不含病毒的培养液的小鼠作为对照组。造模后 7 d、14 d、21 d、28 d取心肌组织作石蜡切片，行苏木精-伊红和 Masson 染色观察病理改变，免疫组化（SABC法）检测心肌MMP-9及Ⅰ、Ⅲ型胶原表达。结果：模型组 MMP-9 于 7 d 可见表达，14 d 达高峰（P<0.05），各时间点 MMP-9 表达均高于对照组（P<0.05）；Ⅰ型胶原于 21 d 表达上调，28 d 表达最高（P<0.05），21 d 和 28 d 表达高于对照组（P<0.05）；Ⅲ 型胶原于28 d 表达上调并高于对照组（P<0.05）；MMP-9 与心肌病理积分呈正相关（r=0.832，P<0.05），与 Ⅰ 型胶原呈负相关（r=-0.791，P<0.05）。结论：VMC 小鼠心肌 MMP-9 在早期即表达增高；MMP-9 可能通过介导 Ⅰ 型胶原的降解代谢而参与 VMC 的病理过程，是导致 VMC 胶原重构和心肌纤维化的重要因素之一。
Abstract
OBJECTIVE: To investigate the dynamic changes of expression of matrix metalloproteinases-9 in myocardium of mice with viral myocarditis (VMC) and its significance in the pathogenesis of viral myocarditis. METHODS: VMC model was prepared by an injection of CVB3 in BALB/C mice. The mice receiving an injection of culture solution without virus were used as the control group. Cardiac tissues were obtained 7, 14, 21 and 28 days after injection and made into paraffin sections. Myocardial histopathologic changes were observed by hematoxylin-eosin staining and Masson staining. The expression of MMP-9, type I collagen and type III collagen in cardiac tissues were quantified by SABC immunohistochemical method. RESULTS: The expression of MMP-9 in the VMC model group was observed on the 7th day, reached a peak on the 14th day, and was significantly higher than that in the control group at all time points (P<0.05). Compared with the control group, the expression of type I collagen in the VMC model group was up-regulated on the 21st day and reached a peak on the 28th day (P<0.05). The expression of type III collagen in the VMC model group was significantly higher than that in the control group on the 28th day (P<0.05). The expression of MMP-9 was positively correlated with myocardial histopathologic scores (r=0.832, P<0.05) and negatively correlated with type I collagen expression (r=-0.791, P<0.05). CONCLUSIONS: MMP-9 is over-expressed at the early stage in VMC mice, and participates in the pathological process of VMC through mediating the degradation metabolism of type I collagen. It may be an important factor that leads to myocardial collagen remodeling and myocardial fibrosis.
关键词
病毒性心肌炎 /
基质金属蛋白酶-9 /
Ⅰ型胶原 /
Ⅲ型胶原 /
小鼠
Key words
Viral myocarditis /
Matrix metalloproteinases-9 /
Type I collagen /
Type III collagen /
Mice
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
［1］Maisch B, Alter P, Karatolius K, Ruppert V, Pankuweit S. The heart in cases of viral, bacterial and parasitic infections［J］. Internist(Berl), 2007, 48(3): 255-267.
［2］Jeserich M, Konstantinides S, Pavlik G, Bode C, Geibel A. Non-invasive imaging in the diagnosis of acute viral myocarditis［J］. Clin Res Cardiol, 2009, 98(12):753-763.
［3］蔡小芳，麦根荣，徐之良. 小儿扩张型心肌病与病毒性心肌炎的临床研究［J］. 中国当代儿科杂志，2004，6(4): 325-327.
［4］Szalay G, Sauter M, Haberland M, Zuegel U, Steinmeyer A, Kandolf R, et al. Osteopontin: a fibrosis-related marker molecule in cardiac remodeling of enterovirus myocarditis in the susceptible host［J］. Circ Res, 2009, 104(7):851-859.
［5］Leipner C, Grün K, Müller A, Buchdunger E, Borsi L, Kosmehl H, et al. Imatinib mesylate attenuates fibrosis in coxsackievirus b3-induced chronic myocarditis［J］. Cardiovasc Res, 2008, 79(1): 118-126.
［6］Heymans S, Pauschinger M, De Palma A, Kallwellis-Opara A, Rutschow S, Swinnen M, et al. Inhibition of urokinase-type plasminogen activator or matrix metalloproteinases prevents cardiac injury and dysfunction during viral myocarditis［J］. Circulation, 2006, 114(6): 565-573.
［7］Nelissen I, Martens E, Van den Steen PE, Proost P, Ronsse I, Opdenakker G. Gelatinase B/matrix metalloproteinase-9 cleaves interferon-beta and is a target for immunotherapy［J］. Brain, 2003, 126(Pt 6): 1371-1381.
［8］Li J, Schwimmbeck PL, Tschope C, Leschka S, Husmann L, Rutschow S, et al. Collagen degradation in a murine myocarditis model: relevance of matrix metalloproteinase in association with inflammatory induction［J］. Cardiovas Res, 2002, 56(2): 235-247.
［9］Rezkalla S, Kloner RA, Khatib G, Smith FE, Khatib R. Effect of metoprolol on the acute phase of coxsackievirus B3 murine myocarditis［J］. J Am Coll Cardiol, 1988, 12(2): 412-414.
［10］孟晓慧，汪翼. 基质金属蛋白酶与小儿心血管疾病的关系［J］. 国外医学儿科学分册，2003，30(3): 144-146.
［11］Meng XH, Wang Y, Zhuang JX, Han XZ, Chen Y, Jin YP, et al. Dynamic changes in myocardial matrix metalloproteinase activity in mice with viral myocarditis［J］. Chin Med J (Engl), 2004, 117(8): 1195-1199.
［12］Sch-nbeck U, Mach F, Libby P. Generation of biologically active IL-1 beta by matrix metalloproteinases: a novel caspase-1-independent pathway of IL-1 beta processing［J］. J Immunol, 1998, 161(7): 3340-3346.
［13］Higuchi H, Hara M, Yamamoto K, Miyamoto T, Kinoshita M, Yamada T, et al. Mast cells play a critical role in the pathogenesis of viral myocarditis［J］. Circulation, 2008, 118(4): 363-372.
［14］Kim JY, Kim WJ, Kim H, Suk K, Lee WH. The stimulation of CD147 induces MMP-9 expression through ERK and NF-kappaB in macrophages: implication for atherosclerosis［J］. Immune Netw, 2009, 9(3): 90-97.
［15］张小佛，于小华，郭富强，李双杰. 病毒性心肌炎患儿血清巨噬细胞移动抑制因子、肿瘤坏死因子-α、白细胞介素-1β表达及其意义［J］. 实用儿科临床杂志，2008，23(1)：35-36.
［16］孟晓慧，汪翼，于永慧，陈瑶，马沛然. 病毒性心肌炎小鼠血管紧张素的改变与心肌胶原的关系［J］. 医学临床研究，2004，21(4): 340-344.
PDF(1530 KB)
