Abstract
Objective: Inflammation which is a response of immune system was demonstrated in many disorders such as atherosclerosis, hypertension, diabetes, cancer and rheumatoid arthritis. Metformin, an oral antidiabetic drug, has anti-inflammatory effect apart from blood glucose regulatory effect. However, the mechanism of its anti-inflammatory effect is not clearly understood. In this study, the effect of metformin on the release of cytokines (TNF-α and IL-6) from LPS stimulated rat mononuclear blood cells was investigated. Methods: Blood samples (5ml) were taken from healthy, male, 8-12 weeks old rats (n=5, 200-250g) through cardiac puncture under general anesthesia into sterile EDTA containing tubes. Monocytes were separated by centrifugation and were resuspended in RPMI 1640 media (3.3±0.2x105 /ml). Cells were then incubated with metformin (2.5 μM, 25 μM, 250 μM) for 2,5 hours followed by addition of LPS (100 ng/ml, 1μg/ml) for further 5 hours. After centrifugation, the supernatant was taken and TNF-α level was measured by ELISA. Results: There was no statistically significant change in the amounts of TNF-α in the LPS + metformin groups compared to the 100 ng/ml LPS group (p>0.05). In LPS+metformin groups, compared to 1 μg/ml LPS, 2.5 μM and 250 μM metformin significantly increased TNF-α levels (p<0.05), while 25 μM metformin did not make a significant difference (p>0.05). The amount of IL-6 was not within measurable range in this study. Conclusion: In summary, metformin increased the amount of released TNF-α rather than decreased in our study.
References
- Mullington JM, Hinze-Selch D, Pollmächer T. Mediators of inflammation and their interaction with sleep: relevance for chronic fatigue syndrome and related conditions. Ann N Y Acad Sci. 2001; 933: 201-10.
- Bienvenu J. (1995) Exploration of cytokines in biological fluids. CR Seances Soc Biol Fil 1995; 189:545-55.
- Armstrong L, Jordan N, Millar A. (1996) Interleukin 10 regulation of TNF-α from human alveolar macrophages and pheripheral blood monocytes. Thorax 1996; 51:143-9.
- Durum SK, Openheim JJ. (1993) Proinflammatory cytokines and immunity. In: Paul WE. Fundemental Immunology. 3rd ed. New York Raven Press Ltd 1993; 801- 835.
- Oppenheim JJ, Ruscetti FW, Faltynek C. (1991) Cytokines. In Sites DP, Terr AT. Basic and Clinical Immunology. 7th ed. California: Appleton and Lange 1991; 78- 101.
- Kishimato T.(1989) The biology of IL-6. Blood 1989; 74: 1-10.
- Dinarello CA.(1993) IL-1 and TNF. In: Lachman PJ, Peters DK, Rosen FS, Walport MJ. Clinical Aspects of Immunology. 5th ed. Boston: Blackwell Scientific Publication 1993; 1: 267-313.
- Lau AS.(1994) Cytokines in the pathogenesis and treatment of infectious diseases. In: Aranoff SC, Hughes WT, Kohl S, Speck WT, Wald ER. Advances in Pediatric Infectious Diseases. Chicago: Mosby Year Book 1994; 211-231.
- Groll AH, Meiser A, Weise M, Rettwitz-Volk W, von Loewenich V, Gussetis ES, Kornhuber B. (1992) Interleukin 6 as early mediator in neonatal sepsis. Pediatr Infect Dis J. 1992 Jun;11(6):496-8.
- Hack CE, De Groot ER, Felt-Bersma RJ, Nuijens JH, Strack Van Schijndel RJ, Eerenberg-Belmer AJ, Thijs LG, Aarden LA. (1989) Increased plasma levels of interleukin-6 in sepsis. Blood. 1989 Oct;74(5):1704-10.
- Rusconi F, Parizzi F, Garlaschi L, Assael BM, Sironi M, Ghezzi P, Mantovani A. (1991) Interleukin 6 activity in infants and children with bacterial meningitis. The Collaborative Study on Meningitis. Pediatr Infect Dis J. 1991 Feb;10(2):117-21.
- Sullivan JS, Kilpatrick L, Castarino AT Jr, Lee SC, Harris MC. (1992) Correlation of plasma cytokine elevations with mortality rate in children with sepsis. J Pediatr 1992; 120: 510-515.
- Alam R.(2003) Chemokines in cell movement and inflammation. Rosenwasser LJ, Borish L. Cytokines in allergic inflammation. Church MK, Shute JK, Sampson AP. Mast cell-derivated mediators. Hirota K, Adolphson CR, Gleich GC. Biology of eosinophils. In: Adkinson NF, Yunginger JW, Busse WW, Bachner BS, Holgate ST, Simons FER. Middleton‟s Allergy. 6th ed. USA: Mosby 2003; 164- 165, 138-139, 205, 314.
- Kriegler M, Perez C, DeFay K, Albert I, Lu SD. (1988) A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988 Apr 8;53(1):45-53.
- Tang P, Hung M-C, Klostergaard J (1996). "Human pro-tumor necrosis factor is a homotrimer". Biochemistry 35 (25):8216–25.doi:10.1021/bi952182t. PMID 8679576
- Bienvenu J.(1995) Exploration of cytokines in biological fluids. CR Seances Soc Biol Fil 1995; 189:545-55
- Armstrong L, Jordan N, Millar A.(1996) Interleukin 10 regulation of TNF-α from human alveolar macrophages and pheripheral blood monocytes. Thorax 1996; 51:143-9
- Adler, A. I., Shaw, E. J., Stokes, T. and Ruiz, F. (2009) Newer agents for blood glucose control in type 2 diabetes: summary of NICE guidance. BMJ 338, b1668
- Nathan, D. M., Buse, J. B., Davidson, M. B., Ferrannini, E., Holman, R. R., Sherwin, R. And Zinman, B. (2009) Medical management of hyperglycemia in type 2 diabetes: a consensus algorithm for the initiation and adjustment of therapy: a consensus statement of the American Diabetes Association and the European Association for the Study of Diabetes. Diabetes Care 32, 193–203
- Selvin, E., Bolen, S., Yeh, H. C., Wiley, C., Wilson, L. M., Marinopoulos, S. S., Feldman, L., Vassy, J., Wilson, R., Bass, E. B. and Brancati, F. L. (2008) Cardiovascular outcomes in trials of oral diabetes medications: a systematic review. Arch. Intern. Med. 168, 2070–2080
- Bergheim I, Luyendyk JP, Steele C, Russell GK, Guo L, Roth RA, Arteel GE. (2006) Metformin prevents endotoxin-induced liver injury after partial hepatectomy. J Pharmacol Exp Ther. 2006 Mar;316(3):1053-61. Epub 2005 Dec 1
- Raso GM, Esposito E, Iacono A, Pacilio M, Cuzzocrea S, Canani RB, Calignano A, Meli R., (2009) Comparative therapeutic effects of metformin and vitamin E in a model of non-alcoholic steatohepatitis in the young rat. Eur J Pharmacol. 2009 Feb 14;604(1-3):125-31. Epub 2008 Dec 14.
- Lin HZ, Yang SQ, Chuckaree C, Kuhajda F, Ronnet G, Diehl AM.(2000) Metformin reverses fatty liver disease in obese, leptin-deficient mice. Nat Med. 2000 Sep;6(9):998-1003.
- Marchesini G, Brizi M,Bianchi G, Tomassetti S, Zoli M, Melchionda N.(2001) Metformin in non-alcoholic steatohepatitis. Lancet. 2001 Sep 15;358(9285):893- 4.
- Jaroslaw W. Zmijewski1,2, Emmanuel Lorne1,3*, Xia Zhao1*, Yuko Tsuruta1, Yonggang Sha1, Gang Liu1, Gene P. Siegal4, and Edward Abraham.(2008) Mitochondrial Respiratory Complex I Regulates Neutrophil Activation and Severity of Lung Injury. Am J Respir Crit Care Med Vol 178. pp 168–179, 2008.
Abstract
Amaç: İmmün sistem yanıtının bir sonucu olan inflamasyon, pek çok hastalıkta ortaya çıkan bir tablodur (ateroskleroz, hipertansiyon, diyabet, kanser, romatoid artrit gibi). Diyabet tedavisinde kullanılan metforminin, kan şekerini düzenleyici etkisinden bağımsız olarak, antienflamatuvar etkisininde olduğu bulunmuştur. Ancak etki mekanizması tam olarak açıklanamamıştır. Bu çalışmada, metforminin, lipopolisakkarit (LPS) ile inkübe edilmiş sıçan monositlerinden proinflamatuar sitokinler olan TNF-α ve IL-6 sentez ve salınımı üzerindeki etkisi araştırıldı. Yöntem: Erkek 8-12 haftalık sağlıklı sıçanlardan (n=5, 200-250 g), genel anestezi (Xylazin 3 mg/kg + Ketamin 90 mg/kg) altında kardiyak puncture yöntemiyle 5 ml kan alındı ve steril EDTA’lı tüplere konuldu. Santrifuj (ficoll 3ml, 400xg, 30dk) yardımıyla mononükleer hücreler ayrıştırıldı. Hücreler RPMI 1640 media ile sulandırıldı (3.3±0.2x105 /ml). Hücreler önce metformin (2.5 μM, 25 μM, 250 μM) ile 2,5 saat inkübe edildi. Bu sürenin sonunda LPS (100 ng/ml, 1μg/ml) eklenerek 5 saat daha inkübe edildi. Daha sonra santrifuj (400xg, 20dk.) yapıldı, süpernatantlar -80oC’de, ELISA yöntemi ile TNF-α ve IL-6 düzeyleri ölçülene kadar saklandı. Bulgular: Hücrelerin deneyler sonunda canlılık oranları %98’in üzerindeydi. TNF-α miktarları arasındaki fark, LPS 100 ng/ml ile, LPS + metformin (2.5μM, 25μM, 250μM) grupları karşılaştırıldığında (2179±359 ve 1613±437, 2915±572, 6059±948 pg/ml), istatistiksel olarak anlamlı bulunmadı (p>0.05). LPS 1μgr/ml ile, LPS+metformin (2,5μM, 25μM, 250μM) grupları karşılaştırıldığında (1752±553 ve 3023±745, 2344±598, 6238±841 pg/ml), 2.5μM (p0.05). IL-6 miktarları ELISA yöntemi ile ölçülecek miktarda bulunmadı. Bu konuda yapılan diğer çalışmalardan elde edilen sonuçlara göre, metforminin proinflamatuar sitokinlerin sentezini inhibe ettiği bulunmuştur. Sonuç: Bu çalışmada, metformin, LPS ile indüklenen sıçan mononükleer hücrelerinden, TNF-α sentez ve salınımını, azaltmaktan ziyade artırmaya yönelik bir etki gösterdi.
References
- Mullington JM, Hinze-Selch D, Pollmächer T. Mediators of inflammation and their interaction with sleep: relevance for chronic fatigue syndrome and related conditions. Ann N Y Acad Sci. 2001; 933: 201-10.
- Bienvenu J. (1995) Exploration of cytokines in biological fluids. CR Seances Soc Biol Fil 1995; 189:545-55.
- Armstrong L, Jordan N, Millar A. (1996) Interleukin 10 regulation of TNF-α from human alveolar macrophages and pheripheral blood monocytes. Thorax 1996; 51:143-9.
- Durum SK, Openheim JJ. (1993) Proinflammatory cytokines and immunity. In: Paul WE. Fundemental Immunology. 3rd ed. New York Raven Press Ltd 1993; 801- 835.
- Oppenheim JJ, Ruscetti FW, Faltynek C. (1991) Cytokines. In Sites DP, Terr AT. Basic and Clinical Immunology. 7th ed. California: Appleton and Lange 1991; 78- 101.
- Kishimato T.(1989) The biology of IL-6. Blood 1989; 74: 1-10.
- Dinarello CA.(1993) IL-1 and TNF. In: Lachman PJ, Peters DK, Rosen FS, Walport MJ. Clinical Aspects of Immunology. 5th ed. Boston: Blackwell Scientific Publication 1993; 1: 267-313.
- Lau AS.(1994) Cytokines in the pathogenesis and treatment of infectious diseases. In: Aranoff SC, Hughes WT, Kohl S, Speck WT, Wald ER. Advances in Pediatric Infectious Diseases. Chicago: Mosby Year Book 1994; 211-231.
- Groll AH, Meiser A, Weise M, Rettwitz-Volk W, von Loewenich V, Gussetis ES, Kornhuber B. (1992) Interleukin 6 as early mediator in neonatal sepsis. Pediatr Infect Dis J. 1992 Jun;11(6):496-8.
- Hack CE, De Groot ER, Felt-Bersma RJ, Nuijens JH, Strack Van Schijndel RJ, Eerenberg-Belmer AJ, Thijs LG, Aarden LA. (1989) Increased plasma levels of interleukin-6 in sepsis. Blood. 1989 Oct;74(5):1704-10.
- Rusconi F, Parizzi F, Garlaschi L, Assael BM, Sironi M, Ghezzi P, Mantovani A. (1991) Interleukin 6 activity in infants and children with bacterial meningitis. The Collaborative Study on Meningitis. Pediatr Infect Dis J. 1991 Feb;10(2):117-21.
- Sullivan JS, Kilpatrick L, Castarino AT Jr, Lee SC, Harris MC. (1992) Correlation of plasma cytokine elevations with mortality rate in children with sepsis. J Pediatr 1992; 120: 510-515.
- Alam R.(2003) Chemokines in cell movement and inflammation. Rosenwasser LJ, Borish L. Cytokines in allergic inflammation. Church MK, Shute JK, Sampson AP. Mast cell-derivated mediators. Hirota K, Adolphson CR, Gleich GC. Biology of eosinophils. In: Adkinson NF, Yunginger JW, Busse WW, Bachner BS, Holgate ST, Simons FER. Middleton‟s Allergy. 6th ed. USA: Mosby 2003; 164- 165, 138-139, 205, 314.
- Kriegler M, Perez C, DeFay K, Albert I, Lu SD. (1988) A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988 Apr 8;53(1):45-53.
- Tang P, Hung M-C, Klostergaard J (1996). "Human pro-tumor necrosis factor is a homotrimer". Biochemistry 35 (25):8216–25.doi:10.1021/bi952182t. PMID 8679576
- Bienvenu J.(1995) Exploration of cytokines in biological fluids. CR Seances Soc Biol Fil 1995; 189:545-55
- Armstrong L, Jordan N, Millar A.(1996) Interleukin 10 regulation of TNF-α from human alveolar macrophages and pheripheral blood monocytes. Thorax 1996; 51:143-9
- Adler, A. I., Shaw, E. J., Stokes, T. and Ruiz, F. (2009) Newer agents for blood glucose control in type 2 diabetes: summary of NICE guidance. BMJ 338, b1668
- Nathan, D. M., Buse, J. B., Davidson, M. B., Ferrannini, E., Holman, R. R., Sherwin, R. And Zinman, B. (2009) Medical management of hyperglycemia in type 2 diabetes: a consensus algorithm for the initiation and adjustment of therapy: a consensus statement of the American Diabetes Association and the European Association for the Study of Diabetes. Diabetes Care 32, 193–203
- Selvin, E., Bolen, S., Yeh, H. C., Wiley, C., Wilson, L. M., Marinopoulos, S. S., Feldman, L., Vassy, J., Wilson, R., Bass, E. B. and Brancati, F. L. (2008) Cardiovascular outcomes in trials of oral diabetes medications: a systematic review. Arch. Intern. Med. 168, 2070–2080
- Bergheim I, Luyendyk JP, Steele C, Russell GK, Guo L, Roth RA, Arteel GE. (2006) Metformin prevents endotoxin-induced liver injury after partial hepatectomy. J Pharmacol Exp Ther. 2006 Mar;316(3):1053-61. Epub 2005 Dec 1
- Raso GM, Esposito E, Iacono A, Pacilio M, Cuzzocrea S, Canani RB, Calignano A, Meli R., (2009) Comparative therapeutic effects of metformin and vitamin E in a model of non-alcoholic steatohepatitis in the young rat. Eur J Pharmacol. 2009 Feb 14;604(1-3):125-31. Epub 2008 Dec 14.
- Lin HZ, Yang SQ, Chuckaree C, Kuhajda F, Ronnet G, Diehl AM.(2000) Metformin reverses fatty liver disease in obese, leptin-deficient mice. Nat Med. 2000 Sep;6(9):998-1003.
- Marchesini G, Brizi M,Bianchi G, Tomassetti S, Zoli M, Melchionda N.(2001) Metformin in non-alcoholic steatohepatitis. Lancet. 2001 Sep 15;358(9285):893- 4.
- Jaroslaw W. Zmijewski1,2, Emmanuel Lorne1,3*, Xia Zhao1*, Yuko Tsuruta1, Yonggang Sha1, Gang Liu1, Gene P. Siegal4, and Edward Abraham.(2008) Mitochondrial Respiratory Complex I Regulates Neutrophil Activation and Severity of Lung Injury. Am J Respir Crit Care Med Vol 178. pp 168–179, 2008.
Details
| Primary Language | English |
|---|---|
| Subjects | Medical Education |
| Journal Section | Research Article |
| Authors | |
| Publication Date | March 29, 2024 |
| Submission Date | January 2, 2024 |
| Acceptance Date | January 22, 2024 |
| Published in Issue | Year 2024 |