- P-ISSN 1010-0695
- E-ISSN 2288-3339
Background: Nitric oxide (NO) is a reactive free radical gas and a messenger molecule. NO has many physiological functions, but excessive NO production induces neurotoxicity. Objective: The present study investigated whether the aqueous extract of Polygala tenuifolia Willdenow possesses a protective effect on NO-induced apoptosis in human neuroblastoma cell line SK-N-MC. Method: For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2- phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and caspase-3 enzyme assay were performed. Result: Sodium nitroprusside (SNP) exposure significantly decreased the viability of cells. The cells treated with SNP exhibited several apoptotic features such as increasing of Bax expression, caspase-3 enzyme activity and inhibiting of Bcl-2 expression. On the other hand, the viability of cells pre-treated with the aqueous extract of Polygala tenuifolia Willdenow was increased dose-dependently. The cells pre-treated for 1 h with the aqueous extract of Polygala tenuifolia Willdenow followed by treatment with SNP showed a decreased occurrence of apoptotic features like decreasing Bax expressions, caspase-3 enzyme activity and increasing Bcl-2 expressions. The aqueous extract of Polygala tenuifolia Willdenow reduced apoptotic cell death in neuroblastoma cell line SK-N-MC through the inhibition of Bax-dependent caspase-3 activation and the increasing of Bcl-2 expression. Conclusion: Based on the present results, it is possible that Polygala tenuifolia Willdenow has therapeutic value for the treatment of a variety of NO-induced brain diseases.
1 Ikeya Y, Takeda S, Tunakawa M, Karakida H, Toda K, Yamaguchi T, et al. Cognitive Improving and Cerebral Protective Effects of Acylated Oligosaccharides in Polygala tenuifolia. Biol Pharm Bull. 2004; 27(7) 1081-5.
2 Park HK, Jeon SG, Kim TB, Kang HR, Chang YS, Kim YK, et al. Occupational asthma and rhinitis induced by a herbal medicine, Wonji (Polygala tenuifolia). J Korean Med Sci. 2005; 20(1):46-9.
3 Schmidt HH, Walter U. NO at work. Cell. 1994; 78:919-25.
4 Yun HY, Dawson VL, Dawson TM. Neurobiology of nitric oxide. Crit Rev Neurobiol. 1996; 10: 291-316.
5 Garthwaite J, Boulton, CL. Nitric oxide signaling in the central nervous system. Annu Rev Physiol. 1995; 57:683-706.
6 Hölscher C. Nitric oxide, the enigmatic neuronal messenger: its role in synaptic plasticity. Trends Neurosci. 1997; 20:298-303.
7 Hawkins RD. NO honey, I don't remember. Neuron. 1996; 16:465-7.
8 Dawson VL, Dawson TM. Nitric oxide in neuronal degeneration. Proc Soc Exp Biol Med. 1996; 211:33-40.
9 Gross SS, Wolin MS. Nitric oxide: pathophysiological mechanisms. Annu Rev Physiol. 1995; 57:737-69.
10 Woodle ES, Kulkarni S. Programmed cell death. Transplantation. 1998; 66:681-91.
11 Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995; 267: 1456-62.
12 Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol. 1980; 68:251-306.
13 Hoetelmans RW. Nuclear partners of Bcl-2: Bax and PML. DNA Cell Biol 2004; 23:351-4.
14 Lowe SW, Ruley HE. Stabilization of the p53 tumor suppressor is induced by adenovirus 5 E1A and accompanies apoptosis. Genes Dev. 1993; 7:535-45.
15 Xiang H, Kinoshita Y, Knudson CM, Korsmeyer SJ, Schwartzkroin PA, Morrison RS. Bax involvement in p53-mediated neuronal cell death. J Neurosci. 1998; 18:1363-73.
16 Budihardjo I, Oliver H, Lutter M, Luo X, Wang X. Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol. 1999; 15:269-90.
17 Korsmeyer SJ. BCL-2 gene family and the regulation of programmed cell death. Cancer Res. 1999; 59:1693-700.
18 Cohen GM. Caspases: the executioners of apoptosis. Biochem J. 1997; 326:1-16.
19 Heales SJR, Bolanos JP, Stewart VC, Brookes PS, Land JM, Clark JB. Nitric oxide, mitochondria and neurological disease. Biochim Biophys Acta. 1999; 1410;215-28.
20 Murphy MP. Nitric oxide and cell death. Biochim Biophys Acta. 1999; 1411:401-14.
21 Bosca L, Hortelano S. Mechanisms of nitric oxide-dependent apoptosis: Involvement of mitochondrial mediators. Cell Signal. 1999; 11:239-44.
22 Terada S, Kumagai T, Yamamoto N, Ogawa A, Ishimura J, Fujita T, et al. Generation of a novel apoptosis-resistant hepatoma cell line. J Biosci Bioeng. 2003; 95(2):146-51.
23 Gu JC, Wang Y, Zhang ZT, Xue JG, Li JS, Zhou YZ. Effects of human interleukin 10 gene transfer on the expression of Bcl-2 Bax and apoptosis of hepatocyte in rats with acute hemorrhagic necrotizing pancreatitis. Chin Med J (Engl). 2005; 118(19):1658-60.
24 Lee MH, Jang MH, Kim EK, Han SW, Cho SY, Kim CJ. Nitric oxide induces apoptosis in mouse C2C12 myoblast cells. J Pharmacol Sci. 2005; 97:369-76.
25 Yung HW, Bal-price AK, Brown GC, Tolkovsky AM. Nitric oxide-induced cell death of cerebrocortical murine astrocytes is mediated through p53- and Bax-dependent pathways. J Neurochem. 2004; 89:812-21.
26 Miyashita T, Reed JC. Tumor suppressor p53 is a direct transcriptional activator of the human Bax gene. Cell. 1995; 80:293-9.
27 McCurrach ME, Connor TM, Knudson CM, Korsmeyer SJ, Lowe SW. Bax-deficiency promotes drug resistance and oncogenic transformation by attenuating p53-dependent apoptosis. Proc Natl Acad Sci USA. 1997; 94:2345-9.
28 Reed JC. Regulation of apoptosis by Bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol. 1995; 7:541-6.