Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2013, Vol. 11 ›› Issue (4): 246-252.doi: 10.3736/jintegrmed2013036

• Research Article • Previous Articles     Next Articles

Extract of buckwheat sprouts scavenges oxidation and inhibits pro-inflammatory mediators in lipopolysaccharide-stimulated macrophages (RAW264.7)

Rajendra Karkia,b, Cheol-Ho Parkc, Dong-Wook Kima()   

  1. a. Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeollanam-do 534-729, South Korea
    b. Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, KansasCity, MO 64108, USA
    c. Department of Bio-Health Technology, Kangwon National University, Chuncheon, Gangwondo 200-701,South Korea
  • Received:2013-04-21 Accepted:2013-06-17 Online:2013-07-10 Published:2013-07-15

Objective

Buckwheat has been considered as a potential source of nutraceutical components on the world market of probiotic foodstuffs. The purpose of this study was to evaluate the effects of tartary buckwheat (Fagopyrum tataricum) sprouts on oxidation and pro-inflammatory mediators.


Methods

The anti-oxidant effects of buckwheat extract (BWE) and rutin were evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH)- and nitric oxide (NO)-scavenging activities, serum peroxidation and chelating assays. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were used to evaluate anti-inflammatory activities of buckwheat and rutin. NO production in LPS-stimulated RAW264.7 cells was determined by using Griess reagent. The expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-κB) p65 subunit in cytosolic and nuclear portions were determined by Western blot analysis. Also, the production of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay.


Results

Inhibitory concentration 50 values for DPPH- and NO-scavenging activities of BWE were 24.97 and 72.54 μg/mL respectively. BWE inhibited serum oxidation and possessed chelating activity. Furthermore, BWE inhibited IL-6 and TNF-α production in LPS-stimulated RAW264.7 cells. Also, BWE inhibited iNOS and COX-2 expression and NF-κB p65 translocation.


Conclusion

Buckwheat sprouts possessed strong antioxidant activity and inhibited production of pro-inflammatory mediators in the applied model systems. Thus, buckwheat can be suggested to be beneficial in inflammatory diseases by inhibiting the free radicals and inflammatory mediators.

Key words: Fagopyrum, Plant extracts, Oxidation, Nitric oxide synthase type II, Cyclooxygenase 2

Figure 1

DPPH-scavenging activity, NO-scavenging activity, metal chelation and serum oxidationA: DPPH-scavenging activity of BWE and rutin. Ascorbic acid was used as the positive control, and values are expressed as mean ± standard deviation, n = 5. B: Nitric oxide-scavenging activity of BWE and rutin. Ascorbic acid was used as the positive control, and values are expressed as mean ± standard deviation, n = 5. C: Chelating activity of BWE and rutin; values are expressed as mean ± standard deviation, n = 5. D: Effects of BWE and rutin on serum lipid oxidation. *P<0.05, **P<0.01, vs control (sample-untreated but CuSO4-treated). Butylated hydroxytoluene was used as positive control; values are expressed as mean ± standard deviation, n = 5. DPPH: 1,1-diphenyl-2-picryl hydrazyl; NO: nitric oxide; BWE: buckwheat extract; BHT: butylated hydroxytoluene; TBARS: thiobarbituric acid reactive substance."

Figure 2

Viability of RAW264.7 cells and NO production RAW264.7 cells were pre-treated with samples for 1 h followed by stimulation of the cells with LPS for additional 24 h. A: The cell viability was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. B: NO production was measured by using Griess reagent. Values are expressed as mean ± standard deviation, n = 3. *P<0.05, **P<0.01, vs control (LPS-treated, sample-untreated). NO: nitric oxide; BWE: buckwheat extract; LPS: lipopolysaccharide."

Figure 3

iNOS and COX-2 expressionRAW264.7 cells were pre-treated with samples for 1 h followed by stimulation of the cells with LPS for additional 24 h. Then, the expression of iNOS and COX-2 was determined by Western blot analysis. b-actin was used for normalization. Values are expressed as mean ± standard deviation, n = 3. *P<0.05 , **P<0.01, vs control (LPS-treated, sample-untreated). iNOS: inducible nitric oxide synthase; COX-2: cyclooxygenase-2; BWE: buckwheat extract; LPS: lipopolysaccharide."

Figure 4

TNF-α and IL-6 production RAW264.7 cells were pre-treated with samples for 1 h followed by stimulation of the cells with LPS for additional 24 h. The levels of TNF-α and IL-6 in supernatant were measured by using enzyme-linked immunosorbent assay. Values are expressed as mean ± standard deviation, n = 3. *P<0.05, **P<0.01, vs control (LPS-treated, sample-untreated). TNF-α: tumor necrosis factor-α; IL-6: interleukin-6; BWE: buckwheat extract; LPS: lipopolysaccharide."

Figure 5

NF-κB translocationRAW264.7 cells were pre-treated with samples for 1 h followed by stimulation of the cells with LPS for additional 30 min. The expression of NF-κB p65 subunit in the cytosolic and nuclear fractions was determined by Western blot analysis. Values are expressed as mean ± standard deviation, n = 3. *P<0.05, **P<0.01, vs control (LPS-treated, sample-untreated). NF-κB: nuclear factor-kappa B; PCNA: proliferating cell nuclear antigen; BWE: buckwheat extract; LPS: lipopolysaccharide."

[1] Ohnishi O . Geographical distribution of allozymes in natural populations of wild Tartary buckwheat[J]. Fagopyrum, 2000,17:29-34
[2] Kim S, Kim SK, Park CH . Introduction and nutritional evaluation of buckwheat sprouts as a new vegetable[J]. Food Res Int, 2004,37(4):319-327
doi: 10.1016/j.foodres.2003.12.008
[3] Kim H, Park KJ, Lim JH . Metabolomic analysis of phenolic compounds in buckwheat(Fagopyrum esculentum M.) sprouts treated with methyl jasmonate[J]. J Agric Food Chem, 2011,59(10):5707-5713
doi: 10.1021/jf200396k
[4] Kim C, Lee WK, No KO, Park SK, Lee MH, Lim SR, Roh SS . Anti-allergic action of buckwheat(Fagopyrum esculentum Moench) grain extract[J]. Int Immunopharmacol, 2003,3(1):129-136
doi: 10.1016/S1567-5769(02)00261-8
[5] Kishore G, Ranjan S, Pandey A, Gupta S . Influence of altitudinal variation on the antioxidant potential of tartary buckwheat of Western Himalaya[J]. Food Sci Biotechnol, 2010,19(5):1355-1363
doi: 10.1007/s10068-010-0193-9
[6] Choi I, Seog H, Park Y, Kim Y, Choi H . Supressive effects of germinated buckwheat on development of fatty liver in mice fed with high-fat diet[J]. Phytomedicine, 2007,14(7-8):563-567
doi: 10.1016/j.phymed.2007.05.002
[7] Lushchak VI . Environmentally induced oxidative stress in aquatic animals[J]. Aquat Toxicol, 2011,101(1):13-30
doi: 10.1016/j.aquatox.2010.10.006
[8] McCall M, Frei B . Can antioxidant vitamins materially reduce oxidative damage in humans? Free Radic Biol Med. 1999; 26(7-8):1034-1053.
doi: 10.1016/S0891-5849(98)00302-5
[9] Pfanzagl B, Tribl F, Koller E , Mslinger T.Homocysteine strongly enhances metal-catalyzed LDL oxidation in the presence of cystine and cysteine[J]. Atherosclerosis, 2003,168(1):39-48
doi: 10.1016/S0021-9150(03)00057-1
[10] Baynes JW . Role of oxidative stress in development of complications in diabetes[J]. Diabetes, 1991,40(4):405-412
[11] Jomova K, Vondrakova D, Lawson M, Valko M . Metals, oxidative stress and neurodegenerative disorders[J]. Mol Cell Biochem, 2010,345(1-2):91-104
doi: 10.1007/s11010-010-0563-x
[12] Zeilhofer H, Brune K . Analgesic strategies beyond the inhibition of cyclooxygenases[J]. Trends Pharmacol Sci, 2006,27(9):467-474
doi: 10.1016/j.tips.2006.07.007
[13] Jia Z, Tang M, Wu J . The determination of flavonoid contents in mulberry and their scavenging effects on superoxides radicals[J]. Food Chem, 1999,64(4):555-559
doi: 10.1016/S0308-8146(98)00102-2
[14] Karki R, Sahi N, Jeon ER, Park YS, Kim DW . Chungtaejeon, a Korean fermented tea, scavenges oxidation and inhibits cytokine induced proliferation and migration of human aortic smooth muscle cells[J]. Plant Foods Hum Nutr, 2011,66(1):27-33
doi: 10.1007/s11130-011-0211-y
[15] Yamaguchi F, Ariga T, Yoshimura Y, Nakazawa H . Anti-oxidative and anti-glycation activity of garcinol from Garcinia indica fruit rind[J]. J Agric Food Chem, 2000,48(2):180-185
doi: 10.1021/jf990845y
[16] Yoon J, Lee YJ, Kim JS, Kang DG, Lee HS . Betulinic acid inhibits high glucose-induced vascular smooth muscle cell proliferation and migration[J]. J Cell Biochem, 2010,111(6):1501-1511
doi: 10.1002/jcb.22880
[17] Fabjan N, Rode J, Kosir IJ, Wang Z, Zhang Z, Kreft I . Tartary buckwheat(Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercetin[J]. J Agric Food Chem, 2003,51(22):6452-6455
doi: 10.1021/jf034543e
[18] Holasova M, Fiedlerova V, Smrcinova H, Orsak M, Lachman J, Vavreinova S . Buckwheat — The source of antioxidant activity in functional foods[J]. Food Res Int, 2002,35(2-3):207-211
doi: 10.1016/S0963-9969(01)00185-5
[19] Fernandez-Orozco R, Piskula MK, Zielinski H, Kozowska H, Frias J, Vidal-Valverde C . Germination as a process to improve the antioxidant capacity of Lupinus angustifolius L. var. Zapaton[J]. Eur Food Res Technol, 2006,223(4):495-502
doi: 10.1007/s00217-005-0229-1
[20] Esterbauer H, Rotheneder M, Striegl G, Waeg G, Ashy A, Sattler W, Jürgens G . Vitamin E and other lipophilic antioxidants protect LDL against oxidation[J]. Eur J Lipid Sci Technol, 1989,91(8):316-324
[21] Russo A, Acquaviva R, Campisi A, Sorrenti V, Di Giacomo C, Virgata G, Barcellona ML, Vanella A . Bioflavonoids as antiradicals, antioxidants and DNA protectors[J]. Cell Biol Toxicol, 2000,16(2):91-98
doi: 10.1023/A:1007685909018
[22] Cooper GJ . Therapeutic potential of copper chelation with triethylenetetramine in managing diabetes mellitus and Alzheimer’s disease[J]. Drugs, 2011,71(10):1281-1320
doi: 10.2165/11591370-000000000-00000
[23] Zhang WJ, Wei H, Frei B . The iron chelator, desferrioxamine, reduces inflammation and atherosclerotic lesion development in experimental mice[J]. Exp Biol Med (Maywood), 2010,235(5):633-641
doi: 10.1258/ebm.2009.009229
[24] Stichtenoth DO , Fr?lich JC.Nitric oxide and inflammatory joint diseases[J]. Br J Rheumatol, 1998,37(3):246-257
doi: 10.1093/rheumatology/37.3.246
[25] Xia Y, Zweier JL . Direct measurement of nitric oxide generation from nitic oxide synthase[J]. Proc Natl Acad Sci U S A, 1997,94(23):12705-12710
doi: 10.1073/pnas.94.23.12705
[26] Turini M , DuBois RN.Cyclooxygenase-2: a therapeutic target[J]. Annu Rev Med, 2002,53:35-57
doi: 10.1146/annurev.med.53.082901.103952
[27] Ding C, Jones G . Anti-interleukin-6 receptor antibody treatment in inflammatory autoimmune diseases[J]. Rev Recent Clin Trials, 2006,1(3):193-200
doi: 10.2174/157488706778250168
[28] Parameswaran N, Patial S . Tumor necrosis factor-a signaling in macrophages[J]. Crit Rev Eukaryot Gene Expr, 2010,20(2):87-103
doi: 10.1615/CritRevEukarGeneExpr.v20.i2
[29] Ishii S, Katsumura T, Shiozuka C, Ooyauchi K, Kawasaki K, Takigawa S, Fukushima T, Tokuji Y, Kinoshita M, Ohnishi M, Kawahara M, Ohba K . Anti-inflammatory effect of buckwheat sprouts in lipopolysaccharide-activated human colon cancer cells and mice[J]. Biosci Biotechnol Biochem, 2008,72(12):3148-3157
doi: 10.1271/bbb.80324
[30] Bhaskar S, Shalini V, Helen A . Quercetin regulates oxidized LDL induced inflammatory changes in human PBMCs by modulating the TLR-NF-κB signaling pathway[J]. Immunobiology, 2011,216(3):367-373
doi: 10.1016/j.imbio.2010.07.011
[1] Wan-jun Guo, Yi Wang, Yu Deng, Lin-yan Cheng, Xin Liu, Ruo-fan Xi, Sheng-jie Zhu, Xin-yi Feng, Liang Hua, Kan Ze, Jian-yong Zhu, Dong-jie Guo, Fu-lun Li. Therapeutic effects of the extract of Sancao Formula, a Chinese herbal compound, on imiquimod-induced psoriasis via cysteine-rich protein 61. Journal of Integrative Medicine, 2022, 20(4): 376-384.
[2] Stefania Lamponi, Anna Maria Aloisi, Claudia Bonechi, Marco Consumi, Alessandro Donati, Gemma Leone, Claudio Rossi, Gabriella Tamasi, Luana Ghiandai, Ersilia Ferrini, Paolo Fiorenzani, Ilaria Ceccarelli, Agnese Magnani. Evaluation of in vitro cell and blood compatibility and in vivo analgesic activity of plant-derived dietary supplements. Journal of Integrative Medicine, 2019, 17(3): 213-220.
[3] Lucky Legbosi Nwidu, Yibala Ibor Oboma. Telfairia occidentalis (Cucurbitaceae) pulp extract mitigates rifampicin-isoniazid-induced hepatotoxicity in an in vivo rat model of oxidative stress. Journal of Integrative Medicine, 2019, 17(1): 46-56.
[4] Supakanya Kumkarnjana, Rutt Suttisri, Ubonthip Nimmannit, Thongchai Koobkokkruad, Chutichot Pattamadilok, Nontima Vardhanabhuti . Anti-adipogenic effect of flavonoids from Chromolaena odorata leaves in 3T3-L1 adipocytes. Journal of Integrative Medicine, 2018, 16(6): 427-434.
[5] Rohitash Jamwal. Bioavailable curcumin formulations: A review of pharmacokinetic studies in healthy volunteers. Journal of Integrative Medicine, 2018, 16(6): 367-374.
[6] Shohini Chakraborty, Nashra Afaq, Neelam Singh, Sukanta Majumdar. Antimicrobial activity of Cannabis sativaThuja orientalis and Psidium guajava leaf extracts against methicillin-resistant Staphylococcus aureus. Journal of Integrative Medicine, 2018, 16(5): 350-357.
[7] Bruno José Martins Da Silva, Amanda Anastácia Pinto Hage, Edilene Oliveira Silva, Ana Paula Drummond Rodrigues. Medicinal plants from the Brazilian Amazonian region and their antileishmanial activity: A review. Journal of Integrative Medicine, 2018, 16(4): 211-222.
[8] Musa Toyin Yakubu, Abdulquadri Funmbi Olutoye. Aphrodisiac activity of aqueous extract of Anthonotha macrophylla P. Beauv. leaves in female Wistar rats. Journal of Integrative Medicine, 2016, 14(5): 400-408.
[9] Nisha Panth, Keshav Raj Paudel, Rajendra Karki. Phytochemical profile and biological activity of Juglans regia. Journal of Integrative Medicine, 2016, 14(5): 359-373.
[10] JiaXia, Li-qunHe, XiaoSu. Interventional mechanisms of herbs or herbal extracts on renal interstitial fibrosis. Journal of Integrative Medicine, 2016, 14(3): 165-173.
[11] Santu Kumar Saha, Sourav Roy, Anisur Rahman Khuda-Bukhsh. Ultra-highly diluted plant extracts of Hydrastis canadensis and Marsdenia condurango induce epigenetic modifications and alter gene expression profiles in HeLa cells in vitro. Journal of Integrative Medicine, 2015, 13(6): 400-411.
[12] Wei Chiang Chan Eric, Kuin Wong Siu. Phytochemistry and pharmacology of ornamental gingers, Hedychium coronarium and Alpinia purpurata: A review. Journal of Integrative Medicine, 2015, 13(6): 368-379.
[13] Rashmi Singh, Shatruhan Sharma, Veena Sharma. Comparative and quantitative analysis of antioxidant and scavenging potential of Indigofera tinctoria Linn. extracts. Journal of Integrative Medicine, 2015, 13(4): 269-278.
[14] Oluwatosin Adaramoye, Bettina Erguen, Olubukola Oyebode, Bianca Nitzsche, Michael Höpfner, Klaus Jung, Anja Rabien. Antioxidant, antiangiogenic and antiproliferative activities of root methanol extract of Calliandra portoricensis in human prostate cancer cells. Journal of Integrative Medicine, 2015, 13(3): 185-193.
[15] Swapnil P. Borse, Bhagyashree B. Kamble. Effects of Ayurvedic Rasayana botanicals on CYP3A4 isoenzyme system. Journal of Integrative Medicine, 2015, 13(3): 165-172.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239