Proof of Concept: Matrix metalloproteinase inhibitor decreases inflammation and improves muscle insulin sensitivity in people with type 2 diabetes

Karen Frankwich, Courtney Tibble, Moises Torres-Gonzalez, Mariah Bonner, Roy Lefkowitz, Matt Tyndall, Geert W Schmid-Schönbein, Francisco Villarreal, Mike Heller, Karen Herbst, Karen Frankwich, Courtney Tibble, Moises Torres-Gonzalez, Mariah Bonner, Roy Lefkowitz, Matt Tyndall, Geert W Schmid-Schönbein, Francisco Villarreal, Mike Heller, Karen Herbst

Abstract

Background: Obesity is a state of subclinical inflammation resulting in loss of function of insulin receptors and decreased insulin sensitivity. Inhibition of the inflammatory enzymes, matrix metalloproteinases (MMPs), for 6 months in rodent models restores insulin receptor function and insulin sensitivity.

Methods: This 12-week double-blind, randomized, placebo (PL)-controlled proof-of-concept study was performed to determine if the MMP inhibitor (MMPI), doxycycline, decreased global markers of inflammation and enhanced muscle insulin sensitivity in obese people with type 2 diabetes (DM2). The study included non-DM2 controls (n = 15), and DM2 subjects randomized to PL (n = 13) or doxycycline 100 mg twice daily (MMPI; n = 11). All participants were evaluated on Day 1; MMPI and PL groups were also evaluated after 84 days of treatment.

Results: There was a significant decrease in inflammatory markers C-reactive protein (P < 0.05) and myeloperoxidase (P = 0.01) in the MMPI but not PL group. The MMPI also significantly increased skeletal muscle activated/total insulin signaling mediators: 3'phosphoinositide kinase-1 (PDK1) (p < 0.03), protein kinase B (PKB/Akt) (p < 0.004), and glycogen synthase kinase 3ß (GSK3ß) (p < 0.03).

Conclusions: This study demonstrated short term treatment of people with diabetes with an MMPI resulted in decreased inflammation and improved insulin sensitivity. Larger, longer studies are warranted to determine if doxycycline can improve glucose control in people with diabetes.

Trial registration: Clinicaltrials.gov NCT01375491.

Figures

Figure 1
Figure 1
MPO and CRP levels for the Control, PL, and MMPI groups. MPO and CRP levels for the Control group at D1, and the PL and MMPI groups at D1 and D84. † P < 0.05 versus Control group. # < 0.05 versus D1 of the MMPI group.
Figure 2
Figure 2
Activation of muscle insulin signaling mediators in PL and MMPI groups. MMPI but not PL improves insulin signaling in muscle. Left side panels, A, B and C are digital representations of western immunoblots of key mediators in the insulin signaling pathway (PDK1, Akt, GSK3β. Amounts of phosphorylated and total PDK-1 (A), Akt (B) and GSK3-β (C) in MMPI and PL participants were analyzed at D1 (labeled B on blots) and D84 (labeled A on blots). Specimens for this analysis were obtained from muscle biopsies of seven consecutive age-matched PL and MMPI subjects. In the right side panel, the data is presented as fold change from D1 to D84 in the percentage of phosphorylated (activated) to total molecules in the PL and MMPI groups.
Figure 3
Figure 3
MMP 2/9 activity in the Control, PL, and MMPI groups. MMP 2/9 activity in the Control group (D1) and PL and MMPI groups (D1 and D84). Activity was measured after mixing plasma with a charge-changing peptide substrate for MMP-2 and MMP-9 followed by electrophoresis (see Methods). *P = 0.001 versus the control group.

References

    1. Curat CA, Wegner V, Sengenes C, Miranville A, Tonus C, Busse R, Bouloumie A. Macrophages in human visceral adipose tissue: increased accumulation in obesity and a source of resistin and visfatin. Diabetologia. 2006;49:744–7. doi: 10.1007/s00125-006-0173-z. Epub 2006 Feb 23.
    1. Chavey C, Mari B, Monthouel MN, Bonnafous S, Anglard P, Van Obberghen E, Tartare-Deckert S. Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation. J Biol Chem. 2003;278:11888–96. doi: 10.1074/jbc.M209196200. Epub 2003 Jan 15.
    1. Derosa G, Ferrari I, D'Angelo A, Tinelli C, Salvadeo SA, Ciccarelli L, Piccinni MN, Gravina A, Ramondetti F, Maffioli P, Cicero AF. Matrix metalloproteinase-2 and −9 levels in obese patients. Endothelium. 2008;15:219–24. doi: 10.1080/10623320802228815.
    1. Catalan V, Gomez-Ambrosi J, Rodriguez A, Ramirez B, Silva C, Rotellar F, Gil MJ, Cienfuegos JA, Salvador J, Fruhbeck G. Increased adipose tissue expression of lipocalin-2 in obesity is related to inflammation and matrix metalloproteinase-2 and metalloproteinase-9 activities in humans. J Mol Med. 2009;87:803–13. doi: 10.1007/s00109-009-0486-8. Epub 2009 May 23.
    1. Andarawewa KL, Rio M-C. In: The Cancer Degradome. Edwards D, Høyer-Hansen G, Blasi F, Slo BF, editor. New York: Springer; 2008. New Insights into MMP function in Adipogenesis; pp. 361–72.
    1. Mieczkowska J, Mosiewicz J, Barud W, Kwasniewski W. Changes in the activity of connective tissue matrix enzymes in the metabolic syndrome. Arch Med Sci. 2011;7:634–41.
    1. Ress C, Tschoner A, Ciardi C, Laimer MW, Engl JW, Sturm W, Weiss H, Tilg H, Ebenbichler CF, Patsch JR, Kaser S. Influence of significant weight loss on serum matrix metalloproteinase (MMP)-7 levels. Eur Cytokine Netw. 2010;21:65–70.
    1. Rodrigues SF, Tran ED, Fortes ZB, Schmid-Schonbein GW. Matrix metalloproteinases cleave the beta2-adrenergic receptor in spontaneously hypertensive rats. Am J Physiol Heart Circ Physiol. 2010;299:H25–35. doi: 10.1152/ajpheart.00620.2009. Epub 2010 Apr 9.
    1. DeLano FA, Schmid-Schonbein GW. Proteinase activity and receptor cleavage: mechanism for insulin resistance in the spontaneously hypertensive rat. Hypertension. 2008;52:415–23. doi: 10.1161/HYPERTENSIONAHA.107.104356.
    1. Lucotti P, Monti LD, Setola E, Galluccio E, Gatti R, Bosi E, Piatti P. Aerobic and resistance training effects compared to aerobic training alone in obese type 2 diabetic patients on diet treatment. Diabetes Res Clin Pract. 2011;94:395–403. doi: 10.1016/j.diabres.2011.08.002. Epub 2011 Sep 3.
    1. Madsen EL, Bruun JM, Skogstrand K, Hougaard DM, Christiansen T, Richelsen B. Long-term weight loss decreases the nontraditional cardiovascular risk factors interleukin-18 and matrix metalloproteinase-9 in obese subjects. Metabolism. 2009;58:946–53. doi: 10.1016/j.metabol.2009.02.031.
    1. Zhang P, Yang YJ, Chen X, Ruan YM, Zhou YW, Tian Y, Chen ZJ. Comparison of doxycycline, losartan, and their combination on the expression of matrix metalloproteinase, tissue inhibitor of matrix metalloproteinase, and collagen remodeling in the noninfarcted myocardium after acute myocardial infarction in rats. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2005;27:53–61.
    1. Li DQ, Shang TY, Kim HS, Solomon A, Lokeshwar BL, Pflugfelder SC. Regulated expression of collagenases MMP-1, -8, and −13 and stromelysins MMP-3, -10, and −11 by human corneal epithelial cells. Invest Ophthalmol Vis Sci. 2003;44:2928–36. doi: 10.1167/iovs.02-0874.
    1. Ryan ME, Usman A, Ramamurthy NS, Golub LM, Greenwald RA. Excessive matrix metalloproteinase activity in diabetes: inhibition by tetracycline analogues with zinc reactivity. Curr Med Chem. 2001;8:305–16. doi: 10.2174/0929867013373598.
    1. Chung AW, Yang HH, Radomski MW, van Breemen C. Long-term doxycycline is more effective than atenolol to prevent thoracic aortic aneurysm in marfan syndrome through the inhibition of matrix metalloproteinase-2 and −9. Circ Res. 2008;102:e73–85. doi: 10.1161/CIRCRESAHA.108.174367.
    1. Hackmann AE, Rubin BG, Sanchez LA, Geraghty PA, Thompson RW, Curci JA. A randomized, placebo-controlled trial of doxycycline after endoluminal aneurysm repair. J Vasc Surg. 2008;48:519–26. doi: 10.1016/j.jvs.2008.03.064. discussion 26.
    1. Sorsa T, Ingman T, Suomalainen K, Halinen S, Saari H, Konttinen YT, Uitto VJ, Golub LM. Cellular source and tetracycline-inhibition of gingival crevicular fluid collagenase of patients with labile diabetes mellitus. J Clin Periodontol. 1992;19:146–9. doi: 10.1111/j.1600-051X.1992.tb00454.x.
    1. Dipl-Pharm SG, Zierath JR. Tackling the Insulin-signalling Cascade. Canadian Journal of Diabetes. 2005;29:239–45.
    1. Unal R, Yao-Borengasser A, Varma V, Rasouli N, Labbate C, Kern PA, Ranganathan G. Matrix metalloproteinase-9 is increased in obese subjects and decreases in response to pioglitazone. J Clin Endocrinol Metabol. 2010;95:2993–3001. doi: 10.1210/jc.2009-2623. Epub 2010 Apr 14.
    1. Friese RS, Rao F, Khandrika S, Thomas B, Ziegler MG, Schmid-Schonbein GW, O'Connor DT. Matrix metalloproteinases: discrete elevations in essential hypertension and hypertensive end-stage renal disease. Clin Exp Hypertens. 2009;31:521–33. doi: 10.3109/10641960802668730.
    1. Lefkowitz RB, Schmid-Schonbein GW, Heller MJ. Whole blood assay for elastase, chymotrypsin, matrix metalloproteinase-2, and matrix metalloproteinase-9 activity. Anal Chem. 2010;82:8251–8. doi: 10.1021/ac101462c.
    1. French LR, Boen JR, Martinez AM, Bushhouse SA, Sprafka JM, Goetz FC. Population-based study of impaired glucose tolerance and type II diabetes in Wadena. Minnesota Diabetes. 1990;39:1131–7.
    1. Abdul-Ghani MA, Matsuda M, Balas B, DeFronzo RA. Muscle and Liver Insulin Resistance Indexes Derived From the Oral Glucose Tolerance Test. Diabetes Care. 2007;30:89–94. doi: 10.2337/dc06-1519.
    1. Antuna-Puente B, Faraj M, Karelis AD, Garrel D, Prud'homme D, Rabasa-Lhoret R, Bastard JP. HOMA or QUICKI: is it useful to test the reproducibility of formulas? Diabetes Metab. 2008;34:294–6. doi: 10.1016/j.diabet.2008.02.001. Epub 2008 May 12.
    1. Nagaretani H, Nakamura T, Funahashi T, Kotani K, Miyanaga M, Tokunaga K, Takahashi M, Nishizawa H, Kishida K, Kuriyama H, Hotta K, Yamashita S, Matsuzawa Y. Visceral fat is a major contributor for multiple risk factor clustering in Japanese men with impaired glucose tolerance. Diabetes Care. 2001;24:2127–33. doi: 10.2337/diacare.24.12.2127.
    1. Phillips DI, Clark PM, Hales CN, Osmond C. Understanding oral glucose tolerance: comparison of glucose or insulin measurements during the oral glucose tolerance test with specific measurements of insulin resistance and insulin secretion. Diabet Med. 1994;11:286–92. doi: 10.1111/j.1464-5491.1994.tb00273.x.
    1. Urbaniak GC, Plous S. Research Randomizer (Version 3.0) [Computer software] 2011. Retrieved on April 22, 2010, from .: Social Psychology Network.
    1. Kelishadi R, Sharifi M, Khosravi A, Adeli K. Relationship between C-reactive protein and atherosclerotic risk factors and oxidative stress markers among young persons 10–18 years old. Clin Chem. 2007;53:456–64. doi: 10.1373/clinchem.2006.073668. Epub 2007 Jan 26.
    1. Roman RM, Camargo PV, Borges FK, Rossini AP, Polanczyk CA. Prognostic value of myeloperoxidase in coronary artery disease: comparison of unstable and stable angina patients. Coron Artery Dis. 2010;21:129–36. doi: 10.1097/MCA.0b013e328333f50d.
    1. Karakas M, Koenig W, Zierer A, Herder C, Rottbauer W, Baumert J, Meisinger C, Thorand B. Myeloperoxidase is Associated with Incident Coronary Heart Disease Independently of Traditional Risk Factors: Results from the MONICA/KORA Augsburg Study. J Intern Med. pp. 1365–2796.
    1. Moldoveanu E, Tanaseanu C, Tanaseanu S, Kosaka T, Manea G, Marta DS, Popescu LM. Plasma markers of endothelial dysfunction in type 2 diabetics. Eur J Intern Med. 2006;17:38–42. doi: 10.1016/j.ejim.2005.09.015.
    1. Golub LM, Lee HM, Lehrer G, Nemiroff A, McNamara TF, Kaplan R, Ramamurthy NS. Minocycline reduces gingival collagenolytic activity during diabetes. Preliminary observations and a proposed new mechanism of action. J Periodontal Res. 1983;18:516–26. doi: 10.1111/j.1600-0765.1983.tb00388.x.
    1. Griffin MO, Jinno M, Miles LA, Villarreal FJ. Reduction of myocardial infarct size by doxycycline: a role for plasmin inhibition. Mol Cell Biochem. 2005;270:1–11. doi: 10.1007/s11010-005-2540-3.
    1. Kraus RL, Pasieczny R, Lariosa-Willingham K, Turner MS, Jiang A, Trauger JW. Antioxidant properties of minocycline: neuroprotection in an oxidative stress assay and direct radical-scavenging activity. J Neurochem. 2005;94:819–27. doi: 10.1111/j.1471-4159.2005.03219.x.
    1. Puyana J, Urena V, Quirce S, Fernandez-Rivas M, Cuevas M, Fraj J. Serum sickness-like syndrome associated with minocycline therapy. Allergy. 1990;45:313–5. doi: 10.1111/j.1398-9995.1990.tb00502.x.
    1. Shapiro LE, Knowles SR, Shear NH. Comparative safety of tetracycline, minocycline, and doxycycline. Arch Dermatol. 1997;133:1224–30. doi: 10.1001/archderm.1997.03890460044005.
    1. Golub LM, Lee HM, Stoner JA, Sorsa T, Reinhardt RA, Wolff MS, Ryan ME, Nummikoski PV, Payne JB. Subantimicrobial-dose doxycycline modulates gingival crevicular fluid biomarkers of periodontitis in postmenopausal osteopenic women. J Periodontol. 2008;79:1409–18. doi: 10.1902/jop.2008.070623.
    1. Daniels JM, Schoorl M, Snijders D, Knol DL, Lutter R, Jansen HM, Boersma WG. Procalcitonin vs C-reactive protein as predictive markers of response to antibiotic therapy in acute exacerbations of COPD. Chest. 2010;138:1108–15. doi: 10.1378/chest.09-2927.
    1. Brown DL, Desai KK, Vakili BA, Nouneh C, Lee HM, Golub LM. Clinical and biochemical results of the metalloproteinase inhibition with subantimicrobial doses of doxycycline to prevent acute coronary syndromes (MIDAS) pilot trial. Arterioscler Thromb Vasc Biol. 2004;24:733–8. doi: 10.1161/01.ATV.0000121571.78696.dc. Epub 2004 Feb 12.
    1. Tran ED, Yang M, Chen A, Delano FA, Murfee WL, Schmid-Schonbein GW. Matrix Metalloproteinase Activity Causes VEGFR-2 Cleavage and Microvascular Rarefaction in Rat Mesentery. Microcirculation. pp. 1549–8719.
    1. Dominguez-Rodriguez A, Abreu-Gonzalez P, Garcia-Gonzalez MJ, Kaski JC. High serum matrix metalloproteinase-9 level predict increased risk of in-hospital cardiac events in patients with type 2 diabetes and ST segment elevation myocardial infarction. Atherosclerosis. 2008;196:365–71. doi: 10.1016/j.atherosclerosis.2006.11.012. Epub 2006 Dec 11.
    1. Lewandowski KC, Banach E, Lewinski A. Concentrations of matrix metalloproteinase 2 and 9 in patients with type 2 diabetes and in non-diabetic controls. Endocr Abstr. 2009;19:P121.

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