By Mark Kaye, DC, Director of Clinical Services, Metagenics Inc.
Nutrigenomic research has yielded the discovery of phytocompounds derived from Humulus lupulus (hops) and berberine with a high degree of predicted safety that may help reduce overexpression of specific matrix metalloproteinases (MMPs) associated with degenerative joint pathologies. MMPs play a key role in the breakdown of collagens that compose the tendons, ligaments, cartilage, and other tissues collectively called the extracellular matrix (ECM).1-17 These novel phytocompounds have been shown to selectively inhibit inflammatory mediators that foster greater production of specific MMPs that lead to cartilage and collagen degradation.
The ECM is the mesh-like extracellular milieu that distinguishes animal tissue and provides support and anchorage for cells. This dynamic structure — composed of collagen, elastin, chondrocytes, osteoblasts, and more — allows for the body’s ability to adapt to gross structural and physiological stressors, such as mechanical loading. The ECM also plays a key role in regulating intercellular communication, and is responsible for the circulation of nutrients and the removal of cellular waste products.12
ECM proteins are regulated by a family of more than 20 zinc-binding MMPs that are further subdivided into groups (e.g., gelatenases, collagenases, stromelysins).11-13,16-18 MMPs are key in the normal turnover activities in ECM components — and are also networked to inflammatory and immune processes.11,13,15,18,19 In lower concentration, MMPs are beneficial to normal growth, tissue repair, and reproduction.11-13,15,17,18 But in higher concentration, MMPs have been implicated in numerous degenerative pathologies.10-15,20-21 Elevated MMP expression has also been associated with tendon pathologies (acute tendon injuries, tendonitis, torn rotator cuffs), degenerative discs, and sites of repeated injury or mechanical strain.22-26
MMP-13, sometimes referred to as collagenase-3, exhibits high activity against type II collagen, the primary collagen found in cartilage, and has been expressed in pathologies associated with excessive ECM degradation.12,13,15,17,27 (MMP-13 also degrades types I, III, IV, X, and XIV collagen, along with aggrecan core protein.)12,13 MMP-13 also plays a role in activation of MMP-2 (gelatinase-A) and MMP-9 (gelatinase-B), which can degrade basement membrane components and are thought to play an important role in final collagen degradation following MMP-13-mediated damage.13,15,18
MMPs are produced by structural cells (fibroblasts, endothelial/epithelial cells) and inflammatory cells (macrophages, lymphocytes, neutrophils, eosinophils).15 The following mediators are pivotal in the regulation of MMP production, expression, and activity:
• Proinflammatory cytokines, such as tumor necrosis factor-a (TNF-?), interleukin-6 (IL-6), oncostatin M (OSM), and interleukin-1? (IL-1?).12,18,19,27-31
• Oxidative stress and reactive oxygen species (ROS), such as hydrogen peroxide.7,9,15,16,32-33
• Transcription factors, such as nuclear factor-?B (NF-?B), activator protein-1 (AP-1), and runt-related transcription factor 2 (RUNX-2).15,18,19,27,30-31,34-35
• Mitogen-transducing signal proteins, including protein kinase C (PKC).15,16,18,31
• Protein kinase B (PKB/Akt) and protein kinase A (PKA).18
• Mitogen-activated protein kinase (MAPK) pathways, including p38 MAPK, the Jun kinase pathway (JNK), and the extracellular signal-regulated kinases 1 and 2 pathway (ERK 1/2).13,15,16,18,27,34-35
• Phosphatidylinositol 3-kinase signaling pathway (PI3K/Akt).18,19,29
• Poly (ADP-ribose) polymerase (PARP).36-38
• Cyclooxygenase-2 (COX-2) and prostaglandins (PGE2).18,34
• Elevated homocysteine.16,32,39
Additionally, MMPs are capable of inducing expression of other MMPs (e.g., MMP-13 can activate MMP-2 and vice versa) and have the ability to produce or modulate precursors to proinflammatory cytokines and transcription factors, thereby contributing to inflammation and further MMP expression.17 Inhibition of signaling pathways, transcription factors, and associated cytokines has therefore been suggested in a growing amount of scientific evidence as a therapeutic approach to conditions associated with MMP overexpression.6-8,10-13,17-18,21,23-25,29-30,37-38,40-42
Natural agents for down-regulating MMP enzymes
The following nutrients and derivatives have been shown to support inhibition of collagen-damaging expression of MMPs:
THIAA. In an extensive screen for kinase activity at the MetaProteomics LLC Nutrigenomics Research Center — the proteomics research facility of Metagenics Inc. —THIAA (tetrahydro iso-alpha acids derived from hops), favorably modulated PKC beta and gamma, two important kinases involved in cell inflammatory processes.1
Berberine. This plant alkaloid has a long history of use in Ayurvedic and traditional Chinese medicine, as well as in a variety of applications in modern clinical use.3,5-9,43-44 Berberine has been shown to down-regulate the activity of MMP-1 (which also acts against type II collagen and activates MMP-2) and MMP-9, as well as modulating the expression or activity associated with ROS, IL-1?, IL-6, NF-?B, AP-1, TNF-?, and kinase pathways.4-9
Selenium. This essential trace element provides defense against ROS and inflammation, and has also been shown to reduce MMP-2 and MMP-9 expression through modulation of ROS and NF-?B, as well as through possible interference with the p38 MAPK pathway.44-46
Zinc. Zinc supplementation may be effective in reducing spontaneous cytokine release and inflammatory activity, as well as support rather than suppress immune response.47-48 In a recent study, an increase in circulating zinc (among subjects who started with low or borderline-normal levels) favorably modulated IL-6 and monocyte chemotactic protein-1 activity (MCP-1, a marker associated with inflammation), as well as natural killer cell activity.48
Biotin. Status of this vitamin may play a role in inflammatory disease. A deficiency in biotin has been suggested to up-regulate TNF-? production.49 One clinical study suggests that biotin supplementation modulates IL-1? and IL-2 expression.50
Niacinamide. Niacinamide has been shown to modulate expression of PARP [Poly (ADP-ribose) polymerase, a protein involved in DNA repair and cellular apoptosis] with a corresponding decrease in activity of transcription factors and cytokines.36-38 An in vitro study demonstrated the effectiveness of niacinamide in inhibiting IL-1?-induced cartilage degeneration.51
Folic acid and vitamins B6 and B12. Folic acid, alone or in combination with vitamins B6 and B12, has been shown to reduce elevated homocysteine levels, associated with inflammatory conditions and the disturbance of collagen synthesis.15-16,39,41,52-54 In a six-week clinical trial with folic acid supplementation, subjects whose elevated homocysteine levels normalized also had a significant reduction in MMP-9 levels.41 Some evidence suggests that homocysteine triggers the ERK 1/2 pathway that regulates MMP-9 expression.16
Nutrigenomic and clinical research
Cell studies conducted by MetaProteomics LLC indicated that a combination formula containing a 1:1 ratio of THIAA to berberine plus biotin, folic ac
id, niacinamide, selenium, zinc, and vitamins B6 and B12 may support the ECM by reducing TNF-?- and IL-1?-induced expression of MMP-13, one of the main MMPs found in cartilage.2
Additionally, the Functional Medicine Research Center — the clinical research arm of Metagenics — measured the efficacy of this combination formula in a small, open-label, case study series that was conducted with offsite practitioners who were asked to select subjects with specific clinical and examination history:
• Patients, for whom bodywork had only been of brief help, previously requiring repeated adjustments
• Patients with active inflammatory challenges, including chronic and acute pain states
• Patients with poor tissue integrity secondary to chronicity of symptoms, fibrosis (fibromyalgia), and hypothyroidism
Subjects (n=12) took two tablets of the THIAA/berberine formula one hour before bodywork, and then one to two tablets as needed three times daily up to 10 tablets per day. Questionnaires were administered at baseline prior to bodywork and administration of formula, and at specific time points: immediately after bodywork and then one hour, six hours, 24 hours, and seven days afterward. Subjects were asked to score the severity of their pain and lack of flexibility using Likert psychometric scales of one to 10. On the pain scale, a score of 10 represented the highest level of pain. On the flexibility scale, a score of one represented the least level of flexibility.
Pain scores over the course of seven days were dramatically decreased (range: two to four) relative to the baseline median score of seven. The median improvement in pain averaged 71 percent immediately after treatment and 70 percent for 24 hours after bodywork and formula administration. The median flexibility score of three at baseline improved immediately after treatment to a score of seven, and was maintained at nearly that level 24 hours later. One week later, the median score for pain remained at a 43 percent reduction from initial score, suggesting lasting benefits from the combination of bodywork and the continuation of nutritional supplementation (Figures 3-5).
Overall tolerance of the product was good. Two subjects noted some gastrointestinal (GI) discomfort after taking the product on an empty stomach, which was addressed by taking the tablets with food. One subject had more persistent GI discomfort, including a presumed episode of gastrointestinal reflux disease.
The collective results from cell line testing and clinical observations suggest that the combination THIAA/berberine formula may offer nutritional support to help modulate cartilage-damaging MMP expression and provide a complement to bodywork to produce noticeable changes in pain and flexibility. This formula may serve as a potential adjunctive approach to therapies specific to the ECM (chiropractic/osteopathic adjustments), as well as inflammatory conditions associated with cartilage degradation, to help enhance mobility.
1 Konda VR, et al. unpublished results.
2 Lamb JJ. THIAA/Berberine Off-Site Study Group. Tripp ML, Bland JS. Unpublished results.
3 Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett. 2004;203(2):127-137.
4 Lee CH, Chen JC, Hsiang CY, Wu SL, Wu HC, Ho TY. Berberine suppressed inflammatory agents-induced interleukin-1 beta and tumor necrosis factor-alpha productions via the inhibition of IkappaB degradation in human lung cells. Pharmacol Res. 2007;56(3):193-201.
5 Kim S, Chung JH. Berberine prevents UV-induced MMP-1 and reduction of type 1 procollagen expression in human dermal fibroblasts. Phytomedicine. 2008 Sep;15(9):749-53.
6 Kim S, Kim Y, Kim JE, Cho KH, Chung JH. Berberine inhibits TPA-induced MMP-9 and IL-6 expression in normal human keratinocytes. Phytomedicine. 2008;15(5):340-347.
7 Lin JP, Yang JS, Wu CC, et al. Berberine induced down-regulation of matrix metallopreoteinase-1, -2, and -9 in human gastric cancer cells (SNU-5) in vitro. In Vivo. 2008;22(2):223-230.
8 Peng PL, Hsieh YS, Wang CJ, Hus JL, Chou FP. Inhibitory effect of berberine on the invasion of human lung cancer cells via decreased productions of urikinase-plasminogen activator and matrix metalloproteinase-2. Toxicol Appl Pharmacol. 2006;214(1):8-15.
9 Wartenberg M, Budde P, De Mareés M, et al. Inhibition of tumor-induced angiogenesis and matrix metalloproteinase expression in confrontation cultures of embryoid bodies and tumor spheroids by plant ingredients used in traditional Chinese medicine. Lab Invest. 2003;83(1):87-98.
10 Burrage PS, Mix KS, Brinckerhoff CE. Matrix metalloproteinases: role in arthritis. Front Biosci. 2006;11:529-543.
11 Malemud CJ. Matrix metalloproteinases (MMPs) in health and disease: an overview. Front Biosci. 2006;11:1(1696-1701).
12 Mix KS, Mengshol JA, Benbo U, Vincenti MP, Sporn MB, Brinckerhoff CE. A synthetic triterpenoid selectively inhibits the induction of matrix metalloproteinases 1 and 13 by inflammatory cytokines. Arthritis Rheum. 2001;44(5):1096-1104.
13 Ravanti L, Kahari VM. Matrix metalloproteinases in wound repair. Int J Mol Med. 2000;6(4):391-407.
14 Smith GN Jr. The role of collagenolytic matrix metalloproteinases in the loss of articular cartilage in osteorarthritis. Front Biosci. 2006;11:3081-3095.
15 Woo CH, Lim JH, Kim JH. Lipopolysaccharide induces matrix metalloproteinase-9 expression via a mitochondrial reactive oxygen species-p38 kinase activator protein-1 pathway in raw 264.7 cells. J Immunol. 2004;173(11):6973-6980.
16 Moshal KS, Sen U, Tyagi N, et al. Regulation of homocysteine–induced MMP-9 by ERK 1/2 pathway. Am J Physiol Cell Physiol. 2006;290(3):C883-C891.
17 Parks WC, Wilson CL, Lopéz-Boado YS. Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol. 2004;4(8):617-629.
18 Reuben PM, Cheung HS. Regulation of matrix metalloproteinase (MMP) gene expression by protein kinases. Front Biosci. 2006;11:1193-1215.
19 Litherland GJ, Dixon C, Lakey RL, et al. Synergistic collagenase expression and cartilage collagenolysis are phosphatidylinositol 3-kinase/Akt signaling dependent. J Biol Chem. 2008;283(21):14221-14229.
20 Gu Z, Kaul M, Yan B, et al. S-nitrosylation of matrix metalloproteinases: signaling pathway to neuronal cell death. Science. 2002;297(5584):1186-1190.
21 Katiyar SK. Matrix metalloproteinases in cancer metastasis: molecular targets for prostate cancer prevention by green tea polyphenols and grape seed proanthocyanidins. Endocr Metab Immune Disord Drug Targets. 2006;6(1):17-24.
22 Riley GP, Curry V, DeGroot J, et al. Matrix metalloproteinase activities and their relationship with collagen remodeling in tendon pathology. Matrix Biol. 2002; 297(5584):1186-1190.
23 Le Maitre CL, Freemont AJ, Hoyland JA. Localization of degradative enzymes and their inhibitors in the degenerate human intervertabral disc. J Pathol. 2004;204(1):47-54.
24 Arnoczky SP, Kavagnino M, Egerbacher M, Caballero O, Gardner K. Matrix metalloproteinase inhibitors of stress-deprived tendons: an in vitro experimental study. Am J Sports Med. 2007;35(5):763-769.
25 Riley G. Chronic tendon pathology: molecular basis and therapeutic implications. Expert Rev Mol Med. 2005;7(5):1-25.
26 Lo IK, Marchuk LL, Hollinshead R, Hart DA, Frank CB. Matrix metallopreoteinase and tissue inhibitor of matrix metalloproteinase
mRNA levels are specifically altered in torn rotator cuff tendons. Am J Sports Med. 2004;32(5):1223-1229.
27 Mengshol JA, Vincenti MP, Brinckerhoff CE. IL-1 induces collagenase-3 (MMP-13) promoter activity in stably transfected chondrocytic cells: requirement for Runx-2 and activation by -38 MAPK and JNK pathways. Nucleic Acids Res. 2001: 29(21):4361-4372.
28 Redlich K, Hayer S, Ricci R, et al. Osteoclasts are essential for TNF-alpha-mediated joint destruction. J Clin Invest. 2002;110(10):1419-1427.
29 O’Keefe RJ, Rosier RN, Teot LA, Stewart JM, Hicks DG. Cytokine and matrix metalloproteinase expression in pigmented villonodular synovitis may mediate bone and cartilage destruction. Iowa Orthop J. 1998;18:26-34.
30 Abramson SB, Amin A. Blocking the effects of IL-1 in rheumatoid arthritis protects bone and cartilage. Rheumatology (Oxford). 2002;41(9):972-980.
31 Estève PO, Chicoine E, Robledo O, et al. Protein kinase C-zeta regulates transcription of the matrix metalloproteinase-9 gene induced by IL-1 and TNF-alpha in glioma cells via NF-kappa B. J Biol Chem. 2002;277-(30):35150-35155.
32 Mujumdar VS, Aru GM, Tyagi SC. Induction of oxidative stress by homocyst(e)ine impairs endothelial function. J Cell Biochem. 2001;82(3):491-500.
33 Nelson KK, Melendez JA. Mitochondrial redox control of matrix metalloproteinases. Free Radic Biol Med. 2004;37(6):768-784.
34 Gomez PF, Pillinger MH, Attur M, et al. Resolution of inflammation: prostaglandin E2 dissociates nuclear trafficking of individual NF-?B subunits (p65, p50) in simulated rheumatoid synovial fibroblasts. J Immunol. 2005;175(10):6924-6930.
35 Wang X, Manner PA, Horner A, Shum L, Tuan RS, Nuckolls G. Regulation of MMP-13 expression by RUNX2 and FGF2 in osteoarthritic cartilage. Osteoarthritis Cartilage. 2004;12(12):963-973.
36 Kao SJ, Liu DD, Su CF, Chen HI. Niacinamide abrogates the organ dysfunction and acute lung injury caused by endotoxin. J Cardiovasc Pharmacol. 2007;50(3):333-342.
37 Ungerstedt JS, Heimersson K, Söderström T, Hansson M. Nicotinamide inhibits endotoxin-induced monocyte tissue factor expression. J Thromb Haemost. 2003;1(12):2554-2560.
38 Garcia S, Bodaño A, Pablos JL, Gómez-Reino JJ, Conde C. Poly(ADP-ribose) polymerase inhibition reduces tumor necrosis factor-induced inflammatory response in rheumatoid synovial fibroblasts. Ann Rheum Dis. 2008;67(5):631-637.
39 Chaussalet M, Lamy E, Foucault-Bertaud A, et al. Homocysteine modulates the proteolytic potential of human vascular endothelial cells. Biochem Biophys Res Commun. 2004;316(1):170-176.
40 Vidal A, Sabatini M, Rolland-Valognes G, Renard P, Madelmont JC, Mounetou E. Synthesis and in vitro evaluation of targeted tetracycline derivatives: effects on inhibition of matrix metalloproteinases. Bioorg Med Chem. 2007;15(6):3468-2374.
41 Holven KB, Halvorsen B, Schulz H, Aukrust P, Ose L, Nenseter MS. Expression of matrix metalloproteinase-9 in mononuclear cells of hyperhomocysteinaemic subjects. Eur J Clin Invest. 2003;33(7):555-560.
42 Takaishi H, Kimura T, Dalal S, Okadda Y, D’Armiento J. Joint diseases and matrix metalloproteinases: a role for MMP-13. Curr Pharm Biotechnol. 2008;9(1):47-54.
43 Berberine. Altern Med Rev. 2000;5(2):175-177.
44 Park JM, Kim A, Oh JH, Chung AS. Methylseleninic acid inhibits PMA-stimulated pro-MMP-2 activation mediated by MT1-MMP expression and further tumor invasion through suppression of NF-kappaB activation. Carcinogenesis. 2007;28(4):837-847.
45 Jiang C, Ganther H, Lu J. Monomethyl selenium—specific inhibition of MMP-2 and VEGF expression: implications for angiogenic switch regulation. Mol Carcinog. 2000;29(4):236-250.
46 Conley SM, Bruhn RL, Morgan PV, Stamer WD. Selenium’s effects on MMP-2 and TIMP-1 secretion by human trabecular meshwork cells. Invest Opthalmol Vis Sci. 2004;45(2):473-479.
47 Kahmann L, Uciechowski P, Warmuth S, et al. Zinc supplementation in the elderly reduces spontaneous inflammatory cytokine release and restores T cell functions. Rejuvenation Res. 2008;11(1):227-237.
48 Mariani E, Neri S, Cattini L, et al. Effect of zinc supplementation in the elderly reduces spontaneous inflammatory cytokine release and restores T cell functions. Rejuvenation Res. 2008;11(1):227-237.
49 Kuroishi T, Endo Y, Muramoto K, Sugawara S. Biotin deficiency up-regulates TNF-alpha production in murine macrophages. J Leukoc Biol. 2008;83(4):912-920.
50 Zempleni J, Helm RM, Mock DM. In vivo biotin supplementation at a pharmacologic dose decreases proliferation rates of human peripheral blood mononuclear cells and cytokine release. J Nutr. 2001;131(5):1479-1484.
51 Duan D, Yang S, Shao Z, Whang H. Xiong X. Protective effect of niacinamide on interleukin-1 beta-induced annulus fibrosus type II collagen degeneration in vitro. J Huazhong Univ Sci Technolog Med Sci. 2007;27(1):68-71.
52 McNulty H, Pentieva K, Hoey L, Ward M. Homocysteine, B-vitamins and CVD. Proc Nutr Soc. 2008;67(2):232-237.
53 Abularrage CJ, Sidawy AN, White PW, et al. Effect of folic acid and vitamins B6 and B12 on microcirculatory vasoreactivity in patients with hyperhomocysteinemia. Vasc Endovascular Surg. 2007;41(4):339-345.
54 Herrmann M, Peter Schmidt J, Umanskaya N, et al. The role of hyperhomocysteinemia as well as folate, vitamin B6 and B12 deficiencies in osteoporosis: a systematic review. Clin Chem Lab Med. 2007;45(12):1621-1632.
This research was provided by Metagenics Inc.