期刊论文详细信息
BMC Complementary and Alternative Medicine
Antioxidant potential, total phenolic and total flavonoid contents of Rhododendron anthopogonoides and its protective effect on hypoxia-induced injury in PC12 cells
Zhengping Jia1  Rongmin Gao1  Pengcheng Fan1  Huiping Ma1  Linlin Jing1 
[1] Department of Pharmacy, Lanzhou General Hospital, Lanzhou Command of CPLA, Lanzhou 730050, People’s Republic of China
关键词: PC12 cells;    Oxidative stress;    Hypoxia;    Antioxidant;    Rhododendron anthopogonoides;   
Others  :  1222529
DOI  :  10.1186/s12906-015-0820-3
 received in 2015-03-05, accepted in 2015-08-12,  发布年份 2015
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【 摘 要 】

Background

Rhododendron anthopogonoides Maxim, a kind of traditional Tibetan medicine, has been used to remove body heat, body detoxification, cough, asthma, stomachic and swelling, eliminate abundant phlegm and inflammatory for a long time. In the present study, the total phenols and total flavonoid contents as well as antioxidative properties of the crude extract and solvent fractions of R. anthopogonoides were determined using seven antioxidant assays. Additionally, the protective effect of the extracts on hypoxia-induced injury in PC12 cells was also investigated.

Methods

The content of total flavonoid and total phenolic was determined by the aluminum colorimetric method and Folin-Ciocalteu assay, respectively. In vitro antioxidant study, the effect of the crude extract and solvent fractions on total antioxidant activity, reducing power, DPPH radical scavenging, ABTS radical scavenging, superoxide radical scavenging, hydroxyl radical scavenging and nitric oxide radical scavenging were examined. The correlation between the phenolic and flavonoid content of the extracts and their antioxidant properties also analyzed. Furthermore, the protective effect of extracts on hypoxia-induced damage on PC12 cells was investigated by cell viability, lactate dehydrogenase (LDH) release, malondialdehyde (MDA) production and the activities of antioxidant enzymes.

Results

Our results showed that ethyl acetate and n-butanol fractions had higher content of phenolics and flavonoid compounds than other fractions. Except ABTS radical assay, n-butanol fraction exhibited the strongest antioxidant activity. While the hexane fraction showed the lowest antioxidant activity. Ethyl acetate also presented excellent antioxidant activity, which was just lower than n-butanol fraction. Significant correlation between the phenolic, flavonoid content of the extract and fractions with antioxidant assay excluding ABTS, OH scavenging assay was observed. Moreover, ethyl acetate and n-butanol fractions showed protective effect in PC12 cell under hypoxia condition, while crude extract and water fraction had no protective effect. In contrast, hexane fraction exhibited strong cytoprotective effect. Further study indicated that pretreatment of PC12 cells with ethyl acetate and n-butanol fractions, prior to hypoxia exposure, significantly increased the survival of cells and the activities of SOD, CAT, GSH-Px and T-AOC, as well as reduced the level of LDH and MDA. The gathered data demonstrated that ethyl acetate and n-butanol fractions were able to protect PC12 cells against hypoxia induced injury through direct free radical scavenging and modulation of endogenous antioxidant enzymes.

Conclusion

These findings suggested that ethyl acetate and n-butanol fractions of R. anthopogonoides had significant antioxidant activity and could prevent PC12 cells against hypoxia-induced injury. So it might be regarded as an excellent source of antioxidants and had great potential to explore as therapeutic agent for preventing hypoxia related sickness in future.

【 授权许可】

   
2015 Jing et al.

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【 参考文献 】
  • [1]Lee K, Roth RA, LaPres JJ. Hypoxia, drug therapy and toxicity. Pharmacol Ther. 2007; 113(2):229-46.
  • [2]Miyamoto L, Yagi Y, Hatano A, Kawazoe K, Ishizawa K, Minakuchi K, et al. Spontaneously hyperactive MEK-Erk pathway mediates paradoxical facilitation of cell proliferation in mild hypoxia. Biochim Biophys Acta. 2015;1850(4):640–46.
  • [3]Hielscher A, Gerecht S. Hypoxia and free radicals: role in tumor progression and the use of engineering-based platforms to address these relationships. Free Radic Biol Med. 2015;79:281–91.
  • [4]Hernansanz-Agustín P, Izquierdo-Álvarez A, Sánchez-Gómez FJ, Ramos E, Villa-Piña T, Lamas S et al.. Acute hypoxia produces a superoxide burst in cells. Free Radic Biol Med. 2014; 71:146-56.
  • [5]Kim YS, Hwang JW, Sung SH, Jeon YJ, Jeong JH, Jeon BT et al.. Antioxidant activity and protective effect of extract of Celosia cristata L. flower on tert-butyl hydroperoxide-induced oxidative hepatotoxicity. Food Chem. 2015; 168:572-9.
  • [6]Khled khoudja N, Boulekbache-Makhlouf L, Madani K. Antioxidant capacity of crude extracts and their solvent fractions of selected Algerian Lamiaceae. Ind Crop Prod. 2014; 52(0):177-82.
  • [7]Shah MA, Bosco SJD, Mir SA. Plant extracts as natural antioxidants in meat and meat products. Meat Sci. 2014; 98(1):21-33.
  • [8]Du L, Miao X, Gao Y, Jia H, Liu K, Liu Y. The protective effects of Trolox-loaded chitosan nanoparticles against hypoxia-mediated cell apoptosis. Nanomedicine. 2014; 10(7):1411-20.
  • [9]Navarro-Yepes J, Zavala-Flores L, Anandhan A, Wang F, Skotak M, Chandra N et al.. Antioxidant gene therapy against neuronal cell death. Pharmacol Ther. 2014; 142(2):206-30.
  • [10]Pandey AK, Patnaik R, Muresanu DF, Sharma A, Sharma HS. Quercetin in hypoxia-induced oxidative stress: novel target for neuroprotection. In: Int Rev Neurobiol, vol. 102. New York: Academic Press; 2012. p. 107–46.
  • [11]Pohanka M. Alzheimer’s disease and related neurodegenerative disorders: implication and counteracting of melatonin. J Appl Biomed. 2011; 9(4):185-96.
  • [12]Li S, Chen G, Zhang C, Wu M, Wu S, Liu Q. Research progress of natural antioxidants in foods for the treatment of diseases. Food Science and Human Wellness, 2014;3(3-4):110–16.
  • [13]Stevanato R, Bertelle M, Fabris S. Photoprotective characteristics of natural antioxidant polyphenols. Regul Toxicol Pharmacol. 2014; 69(1):71-7.
  • [14]Zhong R-Z, Zhou D-W. Oxidative stress and role of natural plant derived antioxidants in animal reproduction. J Integr Agric. 2013; 12(10):1826-38.
  • [15]Liu J, Jia L, Kan J, Jin C-H. In vitro and in vivo antioxidant activity of ethanolic extract of white button mushroom (Agaricus bisporus). Food Chem Toxicol. 2013; 51:310-6.
  • [16]Grice HC. Safety evaluation of butylated hydroxyanisole from the perspective of effects on forestomach and oesophageal squamous epithelium. Food Chem Toxicol. 1988; 26(8):717-23.
  • [17]Yang K, Zhou YX, Wang CF, Du SS, Deng ZW, Liu QZ et al.. Toxicity of Rhododendron anthopogonoides essential oil and its constituent compounds towards Sitophilus zeamais. Molecules. 2011; 16(9):7320-30.
  • [18]Editors ZB. ZhongHua BenCao(Tibetan medicine). Shanghai Scientific and Techincal Publishers, ShangHai; 2002.
  • [19]Popescu R, Kopp B. The genus Rhododendron: an ethnopharmacological and toxicological review. J Ethnopharmacol. 2013; 147(1):42-62.
  • [20]Li XF, Jin HZ, Chen G, Yan SK, Shen YH, Yang M et al.. Study advance on chemical constituents and pharmacological activities of the Tibeta medicine Dali. Tianran Chanwu Yanjiu Yu Kaifa. 2008; 06:1125-8.
  • [21]Dai S-J, Chen R-D, Yu D-Q. Studies on the flavonoid compounds of Rhododendron anthopogonoides. China J Chin Mater Med. 2004; 29(01):48-51.
  • [22]Dai S-J, Yu D-Q. Studies on the flavonoids in stem of Rhododendron anthopogonoide II. China J Chin Mater Med. 2005; 30(23):1830-3.
  • [23]Dong Y-M, Tang X-W, Zhang S-J, Ding S-L, Qiu W, LIu S-X. Study on the chemical compounds in the volati le oils from leaves of rhododendron anthopogonodies Maxim by GC/MS. J Lanzhou Med Coll. 2003; 29(03):15-16+32.
  • [24]Li WW, Hu FZ, Shi ZX. Study on the chemical compound in the volatile oils of the Tibetan medicine Rhododendron anthopogonoides Maxim. J Yunnan Univ. 2004; 26(6A):48-51.
  • [25]Hyun TK, Kim HC, Kim JS. Antioxidant and antidiabetic activity of Thymus quinquecostatus Celak. Ind Crop Prod. 2014; 52:611-6.
  • [26]Hatamnia AA, Abbaspour N, Darvishzadeh R. Antioxidant activity and phenolic profile of different parts of Bene (Pistacia atlantica subsp. kurdica) fruits. Food Chem. 2014; 145:306-11.
  • [27]Vaz JA, Barros L, Martins A, Santos-Buelga C, Vasconcelos MH, Ferreira ICFR. Chemical composition of wild edible mushrooms and antioxidant properties of their water soluble polysaccharidic and ethanolic fractions. Food Chem. 2011; 126(2):610-6.
  • [28]Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999; 26(9–10):1231-7.
  • [29]Halliwell B, Gutteridge JMC, Aruoma OI. The deoxyribose method: a simple “test-tube” assay for determination of rate constants for reactions of hydroxyl radicals. Anal Biochem. 1987; 165(1):215-9.
  • [30]Shukla S, Mehta A, Mehta P, Bajpai VK. Antioxidant ability and total phenolic content of aqueous leaf extract of Stevia rebaudiana Bert. Exp Toxicol Pathol. 2012; 64(7–8):807-11.
  • [31]Sangameswaran B, Balakrishnan BR, Deshraj C, Jayakar B. In vitro antioxidant activity of roots of Thespesia lampas Dalz and Gibs. Pak J Pharm Sci. 2009; 22(4):368-72.
  • [32]Oyaizu M. Studies on products of browning reactions: antioxidative activities of products of browning reaction prepared from glucosamine. Japan J Nutr. 1986; 44:307-15.
  • [33]Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem. 1999; 269(2):337-41.
  • [34]Hwang SJ, Yoon WB, Lee O-H, Cha SJ, Kim JD. Radical-scavenging-linked antioxidant activities of extracts from black chokeberry and blueberry cultivated in Korea. Food Chem. 2014; 146:71-7.
  • [35]Čanadanović-Brunet J, Ćetković G, Šaponjac VT, Stajčić S, Vulić J, Djilas S et al.. Evaluation of phenolic content, antioxidant activity and sensory characteristics of Serbian honey-based product. Ind Crop Prod. 2014; 62:1-7.
  • [36]Leong LP, Shui G. An investigation of antioxidant capacity of fruits in Singapore markets. Food Chem. 2002; 76(1):69-75.
  • [37]Halliwell B, Gutteridge JMC. Role of free radicals and catalytic metal ions in human disease: an overview. In: Lester Packer ANG, editor. Methods enzymol, vol. 186. New York: Academic Press; 1990. p. 1–85.
  • [38]Girgih AT, He R, Hasan FM, Udenigwe CC, Gill TA, Aluko RE. Evaluation of the in vitro antioxidant properties of a cod (Gadus morhua) protein hydrolysate and peptide fractions. Food Chem. 2015; 173:652-9.
  • [39]Yang J, Ou B, Wise ML, Chu Y. In vitro total antioxidant capacity and anti-inflammatory activity of three common oat-derived avenanthramides. Food Chem. 2014; 160:338-45.
  • [40]Taira J, Tsuchida E, Katoh MC, Uehara M, Ogi T. Antioxidant capacity of betacyanins as radical scavengers for peroxyl radical and nitric oxide. Food Chem. 2015; 166:531-6.
  • [41]Kim SJ, Matsushita Y, Fukushima K, Aoki D, Yagami S, Yuk HG et al.. Antioxidant activity of a hydrothermal extract from watermelons. LWT Food Sci Technol. 2014; 59(1):361-8.
  • [42]Lu Y, Yeap Foo L. Antioxidant activities of polyphenols from sage (Salvia officinalis). Food Chem. 2001;75(2):197–202.
  • [43]Dzoyem JP, Eloff JN. Anti-inflammatory, anticholinesterase and antioxidant activity of leaf extracts of twelve plants used traditionally to alleviate pain and inflammation in South Africa. J Ethnopharmacol. 2015; 160:194-201.
  • [44]Luo H, Huang J, Liao WG, Huang QY, Gao YQ. The antioxidant effects of garlic saponins protect PC12 cells from hypoxia-induced damage. Br J Nutr. 2011; 105(8):1164-72.
  • [45]Zhu K-X, Guo X, Guo X-N, Peng W, Zhou H-M. Protective effects of wheat germ protein isolate hydrolysates (WGPIH) against hydrogen peroxide-induced oxidative stress in PC12 cells. Food Res Int. 2013; 53(1):297-303.
  • [46]Si CL, Shen T, Jiang YY, Wu L, Yu GJ, Ren XD et al.. Antioxidant properties and neuroprotective effects of isocampneoside II on hydrogen peroxide-induced oxidative injury in PC12 cells. Food Chem Toxicol. 2013; 59:145-52.
  • [47]Eslami H, Sharifi AM, Rahimi H, Rahati M. Protective effect of telmisartan against oxidative damage induced by high glucose in neuronal PC12 cell. Neurosci Lett. 2014; 558:31-6.
  • [48]Gao Y, Dong C, Yin J, Shen J, Tian J, Li C. Neuroprotective effect of fucoidan on H2O2-induced apoptosis in PC12 cells via activation of PI3K/Akt pathway. Cell Mol Neurobiol. 2012; 32(4):523-9.
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