Background/Aim: Because the pericapillary basement membrane in skeletal muscles of patients with chronic critical limb ischemia (CLI) is thickened, we determined the expression patterns of genes involved in collagen metabolism, using samples from 9 CLI patients, 4 patients with acute limb ischemia and 4 healthy controls. Methods: Gene array analysis, quantitative RT-PCR and semiquantitative grading of immunohistochemical reactivity were performed to determine mRNA/cDNA and protein concentrations. Results: In CLI patients compared to controls, cDNA levels of matrix metalloproteinase (MMP)-9 and MMP-19 were higher, collagen type IV chains A1 and A2, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2 were similar and MMP-2 were lower. On the protein level, MMP-2, MMP-9, MMP-19 and TIMP-1 were more abundantly expressed. In skeletal muscles from patients with acute limb ischemia, cDNA and protein levels of MMP-9, MMP-19, collagen type IV chains, TIMP-1 and TIMP-2 were high. MMP-2 was elevated at the protein but decreased on the cDNA level. Conclusion: Expression of basement membrane components in skeletal muscles of CLI and acute limb ischemia patients is altered, possibly contributing to the pathogenesis of peripheral arterial disease.

Keywords:
Basement remodeling, Limb ischemia, Matrix metalloproteinase, Peripheral arterial disease, Skeletal muscle, Tissue inhibitor of matrix metalloproteinase
1.
Meru AV, Mittra S, Thyagarajan B, Chugh A: Intermittent claudication: an overview. Atherosclerosis 2006;187:221–237.
2.
Baumgartner I, Schainfeld R, Graziani L: Management of peripheral vascular disease. Annu Rev Med 2005;56:249–272.
3.
Cao R, Brakenhielm E, Pawliuk R, Wariaro D, Post MJ, Wahlberg E, Leboulch P, Cao Y: Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat Med 2003;9:604–613.
4.
Tang GL, Chang DS, Sarkar R, Wang R, Messina LM: The effect of gradual or acute arterial occlusion on skeletal muscle blood flow, arteriogenesis, and inflammation in rat hindlimb ischemia. J Vasc Surg 2005;41:312–320.
5.
Baum O, Djonov V, Ganster M, Widmer M, Baumgartner I: Arteriolization of capillaries and FGF-2 upregulation in skeletal muscles of patients with chronic peripheral arterial disease. Microcirculation 2005;12:527–537.
6.
Kalluri R: Basement membranes: structure, assembly and role in tumour angiogenesis. Nat Rev Cancer 2003;3:422–433.
7.
Kovanen V: Intramuscular extracellular matrix: complex environment of muscle cells. Exerc Sport Sci Rev 2002;30:20–25.
8.
Visse R, Nagase H: Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 2003;92:827–839.
9.
Galis ZS, Khatri JJ: Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly. Circ Res 2002;90:251–262.
10.
Brown MD, Hudlicka O: Modulation of physiological angiogenesis in skeletal muscle by mechanical forces: involvement of VEGF and metalloproteinases. Angiogenesis 2003;6:1–14.
11.
Carmeli E, Moas M, Reznick AZ, Coleman R: Matrix metalloproteinases and skeletal muscle: a brief review. Muscle Nerve 2004;29:191–197.
12.
Second European consensus document on chronic critical leg ischemia. Circulation 1991;84(suppl IV):IV1–IV26.
13.
Gehlenborg N, Dietzsch J, Nieselt K: A framework for visualization of microarray data and integrated meta information. Inf Vis 2005;4:164–175.
14.
Sadowski T, Dietrich S, Muller M, Havlickova B, Schunck M, Proksch E, Muller MS, Sedlacek R: Matrix metalloproteinase-19 expression in normal and diseased skin: dysregulation by epidermal proliferation. J Invest Dermatol 2003;121:989–996.
15.
McLennan IS: Degenerating and regenerating skeletal muscles contain several subpopulations of macrophages with distinct spatial and temporal distributions. J Anat 1996;188:17–28.
16.
Breitling R, Armengaud P, Amtmann A, Herzyk P: Rank products: a simple, yet powerful, new method to detect differentially regulated genes in replicated microarray experiments. FEBS Lett 2004;573:83–92.
17.
Tuomisto TT, Rissanen TT, Vajanto I, Korkeela A, Rutanen J, Yla-Herttuala S: HIF-VEGF-VEGFR-2, TNF-α and IGF pathways are upregulated in critical human skeletal muscle ischemia as studied with DNA array. Atherosclerosis 2004;174:111–120.
18.
Feingold KR, Browner WS, Siperstein MD: Prospective studies of muscle capillary basement membrane width in prediabetics. J Clin Endocrinol Metab 1989;69:784–789.
19.
Williamson JR, Hoffmann PL, Kohrt WM, Spina RJ, Coggan AR, Holloszy O: Endurance exercise training decreases capillary basement membrane width in older nondiabetic and diabetic adults. J Appl Physiol 1996;80:747–753.
20.
Dalton SJ, Mitchell DC, Whiting CV, Tarlton JF: Abnormal extracellular matrix metabolism in chronically ischemic skin: a mechanism for dermal failure in leg ulcers. J Invest Dermatol 2005;125:373–379.
21.
Lee CW, Stabile E, Kinnaird T, Shou M, Devaney JM, Epstein SE, Burnett MS: Temporal patterns of gene expression after acute hindlimb ischemia in mice: insights into the genomic program for collateral vessel development. J Am Coll Cardiol 2004;43:474–482.
22.
Roach DM, Fitridge RA, Laws PE, Millard SH, Varelias A, Cowled PA: Up-regulation of MMP-2 and MMP-9 leads to degradation of type IV collagen during skeletal muscle reperfusion injury; protection by the MMP inhibitor, doxycycline. Eur J Vasc Endovasc Surg 2002;23:260–269.
23.
Perhonen M, Wang W, Han X, Ruskoaho H, Takala TE: Right ventricular collagen type III and IV gene expression increases during early phases of endurance training in hypobaric hypoxic condition. Basic Res Cardiol 1997;92:299–309.
24.
Tayebjee MH, Tan KT, MacFadyen RJ, Lip GY: Abnormal circulating levels of metalloprotease 9 and its tissue inhibitor 1 in angiographically proven peripheral arterial disease: relationship to disease severity. J Intern Med 2005;257:110–116.
25.
Signorelli SS, Malaponte G, Libra M, Di Pino L, Celotta G, Bevelacqua V, Petrina M, Nicotra GS, Indelicato M, Navolanic PM, Pennisi G, Mazzarino MC: Plasma levels and zymographic activities of matrix metalloproteinases 2 and 9 in type II diabetics with peripheral arterial disease. Vasc Med 2005;10:1–6.
26.
Blankenberg S, Rupprecht HJ, Poirier O, Bickel C, Smieja M, Hafner G, Meyer J, Cambien F, Tiret L: Plasma concentrations and genetic variation of matrix metalloproteinase 9 and prognosis of patients with cardiovascular disease. Circulation 2003;107:1579–1585.
27.
Kolb C, Mauch S, Krawinkel U, Sedlacek R: Matrix metalloproteinase-19 in capillary endothelial cells: expression in acutely, but not in chronically, inflamed synovium. Exp Cell Res 1999;250:122–130.
28.
Puhakka HL, Turunen P, Rutanen J, Hiltunen MO, Turunen MP, Yla-Herttuala S: Tissue inhibitor of metalloproteinase 1 adenoviral gene therapy alone is equally effective in reducing restenosis as combination gene therapy in a rabbit restenosis model. J Vasc Res 2005;42:361–367.
29.
Bustin SA: Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 2002;29:23–39.
30.
Mahoney DJ, Carey K, Fu MH, Snow R, Cameron-Smith D, Parise G, Tarnopolsky MA: Real-time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise. Physiol Genomics 2004;18:226–231.
31.
Milkiewicz M, Haas TL: Effect of mechanical stretch on HIF-1α and MMP-2 expression in capillaries isolated from overloaded skeletal muscles: laser capture microdissection study. Am J Physiol Heart Circ Physiol 2005;289:H1315–H1320.
32.
Rajashekhar G, Willuweit A, Patterson CE, Sun P, Hilbig A, Breier G, Helisch A, Clauss M: Continuous endothelial cell activation increases angiogenesis: evidence for the direct role of endothelium linking angiogenesis and inflammation. J Vasc Res 2006;43:193–204.
33.
Hoppeler H, Hudlicka O, Uhlmann E, Claassen H: Capillaries and mitochondria in rat muscles with limited blood supply: a report on a morphometric study in progress; in Okyayuz-Bakluti I, Hudlicka O (eds): Muscle Ischaemia: Functional and Metabolic Aspects. Munich, Wolf und Sohn, 1988, pp 89–98.
34.
Yu J, deMuinck ED, Zhuang Z, Drinane M, Kauser K, Rubanyi GM, Qian HS, Murata T, Escalante B, Sessa WC: Endothelial nitric oxide synthase is critical for ischemic remodeling, mural cell recruitment, and blood flow reserve. Proc Natl Acad Sci USA 2005;102:10999–11004.
35.
Brown MD, Kelsall CJ, Milkiewicz M, Anderson S, Hudlicka O: A new model of peripheral arterial disease: sustained impairment of nutritive microcirculation and its recovery by chronic electrical stimulation. Microcirculation 2005;12:373–381.
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2007
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