Tetramethylpyrazine Attenuates Adriamycin-Induced Apoptotic Injury in Rat Renal Tubular Cells NRK-52E

Abstract

Tetramethylpyrazine (TMP), a compound purified from Rhizoma Ligustici, is a widely used active ingredient in Chinese herbal medicine to treat cardiovascular diseases on account of its vasodilatory actions and antiplatelet activity. Studies have shown that TMP can remove oxygen free radicals and protect rat kidney from ischemia-reperfusion injury. In addition, adriamycin-induced nephrosis in rats is commonly used in pharmacological studies of human chronic renal diseases. Apoptosis of renal tubular cells has been reported in adriamycin-treated rats. To examine the therapeutic potential of TMP on chronic progressive renal diseases, adriamycin-induced injury in rat renal tubular cells NRK-52E has been used to monitor its protective effect. In TUNEL staining, TMP showed a dose-dependent protective effect against adriamycin-induced apoptosis in NRK-52E cells. Pretreatment of the cells with 10 or 100 µM of TMP effectively decreased the reactive oxygen species (ROS) formation induced by adriamycin, as measured in fluorescent assays. TMP was found to reduce the adriamycin-stimulated activities of caspase-3, caspase-8 and caspase-9, inhibit adriamycin-induced release of cytochrome c, and elevate the expression of Bcl-x_L. TMP was also able to inhibit the death receptor signaling pathway and suppress the activation of transcription factor NF-κB in adriamycin-treated NRK-52E cells. Based on the results of this study, we suggest that TMP can attenuate adriamycin-induced oxidative stress and apoptotic injury in NRK-52E cells, and that it may have therapeutic potential for patients with renal diseases.

Abbreviations

TMP: tetramethylpyrazine

LDH: lactate dehydrogenase

ROS: reactive oxygen species

DCF: 2′,7′-dichlorofluorescein

TNF-α: tumor necrosis factor-α

TUNEL: terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling

Key words

Tetramethylpyrazine (TMP) - adriamycin - renal tubular cells - NRK-52E - apoptosis

References

• 1 Liao F. Herbs of activating blood circulation to remove blood stasis.  Clin Hemorheol Microcirc. 2000;  23 127-31
• 2 Liu S Y, Sylvester D M. Antiplatelet activity of tetramethylpyrazine.  Thromb Res. 1994;  75 51-62
• 3 Kwan C Y. Plant-derived drugs acting on cellular Ca²⁺ mobilization in vascular smooth muscle: tetramethylpyrazine and tetrandrine.  Stem Cells. 1994;  12 64-7
• 4 Huang X, Ren P, Wen A D, Wang L L, Zhang L, Gao F. Pharmacokinetics of traditional Chinese syndrome and recipe: a hypothesis and its verification (I).  World J Gastroenterol. 2000;  6 384-91
• 5 Li M, Handa S, Ikeda Y, Goto S. Specific inhibiting characteristics of tetramethylpyrazine, one of the active ingredients of the Chinese herbal medicine ”Chuanxiong,” on platelet thrombus formation under high shear rates.  Thromb Res. 2001;  104 15-28
• 6 Liu C F, Lin C C, Ng L T, Lin S C. Protection by tetramethylpyrazine in acute absolute ethanol-induced gastric lesions.  J Biomed Sci. 2002;  9 395-400
• 7 Liu C F, Lin M H, Lin C C, Chang H W, Lin S C. Protective effect of tetramethylpyrazine on absolute ethanol-induced renal toxicity in mice.  J Biomed Sci. 2002;  9 299-302
• 8 Sun L, Li Y, Shi J, Wang X. Protective effects of ligustrazine on ischemia-reperfusion injury in rat kidneys.  Microsurgery. 2002;  22 343-6
• 9 Feng L, Xiong Y, Cheng F, Zhang L, Li S, Li Y. Effect of ligustrazine on ischemia-reperfusion injury in murine kidney.  Transplant Proc. 2004;  36 1949-51
• 10 Okuda S, Oh Y, Tsuruda H, Onoyama K, Fujimi S, Fujishima M. Adriamycin-induced nephropathy as a model of chronic progressive glomerular disease.  Kidney Int. 1986;  29 502-10
• 11 Bertani T, Poggi A, Pozzoni R, Delaini F, Sacchi G, Thoua Y. et al . Adriamycin-induced nephrotic syndrome in rats: sequence of pathologic events.  Lab Invest. 1982;  46 16-23
• 12 Muller I, Niethammer D, Bruchelt G. Anthracycline-derived chemotherapeutics in apoptosis and free radical cytotoxicity.  Int J Mol Med. 1998;  1 491-4
• 13 Zhang J, Clark JR J r, Herman E H, Ferrans V J. Doxorubicin-induced apoptosis in spontaneously hypertensive rats: differential effects in heart, kidney and intestine, and inhibition by ICRF-187.  J Mol Cell Cardiol. 1996;  28 1931-43
• 14 Khan S, Cleveland R P, Koch C J, Schelling J R. Hypoxia induces renal tubular epithelial cell apoptosis in chronic renal disease.  Lab Invest. 1999;  79 1089-99
• 15 Schelling J R, Nkemere N, Kopp J B, Cleveland R P. Fas-dependent fratricidal apoptosis is a mechanism of tubular epithelial cell deletion in chronic renal failure.  Lab Invest. 1998;  78 813-24
• 16 Singal P K, Li T, Kumar D, Danelisen I, Iliskovic N. Adriamycin-induced heart failure: mechanism and modulation.  Mol Cell Biochem. 2000;  207 77-86
• 17 Kotamraju S, Konorev E A, Joseph J, Kalyanaraman B. Doxorubicin-induced apoptosis in endothelial cells and cardiomyocytes is ameliorated by nitrone spin traps and ebselen. Role of reactive oxygen and nitrogen species.  J Biol Chem. 2000;  275 33 585-92
• 18 Sawyer D B, Fukazawa R, Arstall M A, Kelly R A. Daunorubicin-induced apoptosis in rat cardiac myocytes is inhibited by dexrazoxane.  Circ Res. 1999;  84 257-65
• 19 Jiang X, Wang X. Cytochrome c-mediated apoptosis.  Annu Rev Biochem. 2004;  73 87-106
• 20 Yang J, Liu X, Bhalla K, Kim C N, Ibrado A M, Cai J. et al . Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked.  Science. 1997;  275 1129-32
• 21 Wang S, Kotamraju S, Konorev E, Kalivendi S, Joseph J, Kalyanaraman B. Activation of nuclear factor-kappaB during doxorubicin-induced apoptosis in endothelial cells and myocytes is pro-apoptotic: the role of hydrogen peroxide.  Biochem J. 2002;  367 729-40
• 22 Chen C H, Lin H, Hsu Y H, Sue Y M, Cheng T H, Chan P. et al . The protective effect of prostacyclin on adriamycin-induced apoptosis in rat renal tubular cells.  Eur J Pharmacol. 2006;  529 8-15
• 23 Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD.  Nature. 1998;  391 43-50
• 24 Sakahira H, Enari M, Nagata S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis.  Nature. 1998;  391 96-9
• 25 Liu Z G. Molecular mechanism of TNF signaling and beyond.  Cell Res. 2005;  15 24-7
• 26 Arlt A, Vorndamm J, Breitenbroich M, Folsch U R, Kalthoff H, Schmidt W E. et al . Inhibition of NF-kappaB sensitizes human pancreatic carcinoma cells to apoptosis induced by etoposide (VP16) or doxorubicin.  Oncogene. 2001;  20 859-68
• 27 Somerville L, Cory J G. Enhanced roscovitine-induced apoptosis is mediated by a caspase-3-like activity in deoxyadenosine-resistant mouse leukemia L1210 cells.  Anticancer Res. 2000;  20 3347-55
• 28 Chan H, Bartos D P, Owen-Schaub L B. Activation-dependent transcriptional regulation of the human Fas promoter requires NF-kappaB p50-p65 recruitment.  Mol Cell Biol. 1999;  19 2098-108
• 29 Qin Z H, Chen R W, Wang Y, Nakai M, Chuang D M, Chase T N. Nuclear factor kappaB nuclear translocation upregulates c-Myc and p53 expression during NMDA receptor-mediated apoptosis in rat striatum.  J Neurosci. 1999;  19 4023-33