Arzneimittelforschung 2010; 60(10): 621-626
DOI: 10.1055/s-0031-1296336
Liver Therapeutics
Editio Cantor Verlag Aulendorf (Germany)

Inhibition by folic acid of tumor necrosis factor alpha and apoptosis following normothermic ischemia-reperfusion

Pronobesh Chattopadhyay
1   Cellular and Microbiology Laboratory, College of Pharmacy, Institute of Foreign Trade and Management, Moradabad, Uttar Pradesh, India
2   Birla Institute of Technology and Sciences, Pilani, Rajasthan, India
,
Aran Kumar Wahi
1   Cellular and Microbiology Laboratory, College of Pharmacy, Institute of Foreign Trade and Management, Moradabad, Uttar Pradesh, India
› Author Affiliations
Further Information

Publication History

Publication Date:
03 December 2011 (online)

Abstract

Folate is necessary for the production and maintenance of new cells and important during periods of rapid cell division and growth. Tumor necrosis factor-a (TNF-α) is known as a stimulant of apoptotic cell death. The present study was aimed to evaluate the efficacy of folic acid (CAS 59-30-3) in prevention of apoptosis by inhibiting TNF-α action in ischemia-reperfusion (I/R) induced liver injury. Eighteen Wistar rats were subjected to 1 h of hepatic ischemia followed by 3 h reperfusion and were divided into sham-operated control Group (I) (n = 6), ischemia and reperfusion group administered 0.9% saline (5 ml/kg, p.o.) for 7 days (II) (n = 6), folic acid treated group (1 mg/kg body weight/treated daily by oral route for 7 days before induced ischemia-reperfusion maneuver) (III) (n = 6). Hepatic damage in rats was assessed in terms of serum alanine transaminases and aspartate transaminases. TNF-α concentration was measured in serum by enzyme linked immuno assay. Necrosis and apoptosis were measured by flow cytometry and fluorescence microscopy. Apoptotic marker Bcl-2 gene expression was measured by reverse transcriptase polymerase chain reaction (RTPCR) and Western Blot Analysis. Pathological changes were measured by transmission electron microscopy (TEM).

Serum alanine transaminase (ALT), aspartate transaminase (AST) and TNF-α concentration increased significantly (p < 0.05) in rats with I/R induced injury as compared to sham operated control rats. Pretreatment with folic acid effectively counteracted the alternation in hepatic enzymes. TEM, expression of Bcl-2 protein and flow cytometry studies confirmed the restoration of cellular normalcy and accredits the cytoprotective role of folic acid against I/R induced hepatic injury. The present study demonstrated that elevated TNF-α activity directly related to apoptosis and folic acid inhibits apoptosis by inhibiting the action of TNF-α.

 
  • References

  • 1 Fondevilia C, Busuttil RW, Kupiec-Weglinski JW. Hepatic ischemia/reperfusion injury: a fresh look. Exp Mol Pathol. 2003; 74: 86-93
  • 2 Jaeschke H, Farhood A. Neutrophils and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver. Am J Physiol. 1991; 260: G355-62
  • 3 El-Ghoneimi A, Cursio R, Schmid-Alliana A, Tovey M, Lasfar A, Michiels JF et al. Inhibition of Tumor necrosis factor alpha gene transcription by pentoxifylline reduces nor-mothermic liver ischemia-reperfusion injury in rats. Transplant Proc. 2007; 39: 176-4
  • 4 Blount BC, Mack MM, Wehr CM, MacGregor JT, Hiatt RA, Wang G et al. Folate deficiency causes uracil misincor-poration into human DNA and chromosome breakage: implications for cancer and neuronal damage. Proc Natl Acad Sci USA. 1997; 94: 3290-95
  • 5 Matthias E, Michael A, Inna K, Detlev B, Andreas M, Karen G. Folate deficiency increases post ischemic brain injury. Stroke. 2005; 36: 321-325
  • 6 Chattopadhyay P, Sharma AK, Wahi AK. Folic acid protects hepatobiliary function in ischemic reperfusion of rat liver. Ind J Gastroenterol. 2007; 26: 95-6
  • 7 Chirgwin JM, Przybyla AE, Macdonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979; 18: 5294-9
  • 8 Kreamer BI, Staecker JL, Sawada N, Sattler GL, Hsia MT, Pitot HC. Use of low-speed, iso-density percoll centrifugation method to increase the viability of isolated rat hepato-cytes preparations. In Vitro Cell Dev Biol. 1986; 22: 201-11
  • 9 Colletti LM, Remick DG, Burtch GD, Kunkel SL, Strieter RM et al. Role of tumor necrosis factor-alpha in the pathophysiologic alterations after hepatic ischemia/reperfusion injury in the rat. J Clin Invest. 1990; 85: 1936-43
  • 10 Suzuki S, Toledo-Pereyra LH. Interleukin 1 and tumor necrosis factor production as the initial stimulants of liver ischemia and reperfusion injury. J Surg Res. 1994; 57: 253-8
  • 11 Colletti LM, Kunkel SL, Walz A, Burdick MD, Kunkel RG, Campbell DA. The role of cytokine networks in the local liver injury following hepatic ischemia/reperfusion in the rat. Hepatology. 1996; 23: 506-14
  • 12 Goto M, Takei Y, Kawano S, Tsuji S, Fukui H, Fushimi H et al. Tumor necrosis factor and endotoxin in the pathogenesis of liver and pulmonary injuries after orthotropic liver transplantation in the rat. Hepatology. 1992; 16: 487-93
  • 13 Schlayer HJ, Laaff H, Peters T. Involvement of tumor necrosis factor in endotoxin-triggered neutrophil adherence to sinusoidal cells of mouse liver and its modulation in acute phase. J Hepatol. 1998; 7: 239-49
  • 14 Bevilacqua MP, Pober JS, Majeau GR, Fiers W, Cotran RS, Gimbrone MA. Recombinant tumor necrosis factor induces pro-coagulant activity in cultured human vascular endothelium. Proc Natl Acad Sei USA. 1998; 83: 4533-7
  • 15 Cursio R, Gugenheim J, Panata-Ferrari P, Lasfar A, Tovey M, Chastanet S. Improvement of normothermic rat liver ischemia/reperfusion by muramyl dipeptide. J Surg Res. 1998; 80: 339-44
  • 16 Leist M, Gantner F, Bohlinger I, Germann PG, Tieqs G, Wendel A. Murine hepatocyte apoptosis induced in vitro and in vivo by TNF-alpha requires transcriptional arrest. J Immunol. 1994; 153: 1778-88
  • 17 Chen Z, Chua CC, Ho YS, Hamdy RC, Chua BHL. Overex-pression of Bcl-2 attenuates apoptosis and protects against myocardial I/R injury in transgenic mice. Am J Physiol. 2001; 280 H 2312-20
  • 18 Goulian M, Bleile B, Tseng BY. Methotrexate-induced mis-incorporation of uracil into DNA. Proc Natl Acad Sci USA. 1980; 77: 1956-60
  • 19 Jaeschke H. Cellular adhesion molecules: regulation and functional significance in the pathogenesis of liver diseases. Am J Physiol Gastrointest Liver Physiol. 1997; 273: G602-11
  • 20 Borman LS, Branda RF. Nutritional folate deficiency in Chinese hamster ovary cells. I. Characterization of the pleiotropic response and its modulation by nucleic acid precursors. J Cell Physiol. 1989; 140: 335-43
  • 21 Branda RF, Blickensderfer DB. Folate deficiency increases genetic damage caused by alkylating agents and γ-irradiation in Chinese hamster ovary cells. Cancer Res. 1993; 53: 540-8
  • 22 Zhang X, Chen S, Li L, Wang Q, Le W. Folic acid protects motor neurons against the increased homocysteine, inflammation and apoptosis in SOD1 G93A transgenic mice. Neuropharmacology. 2008; 54: 1112-9