[논문]Rhus verniciflua Stokes Extract and Its Flavonoids Protect PC-12 Cells against H2O2-Induced Cytotoxicity

Nam, Tae Gyu; Lee, Bong Han; Choi, Hyo-Kyoung; Mansur, Ahmad Rois; Lee, Sang Gil; Kim, Dae-Ok

doi:10.4014/jmb.1612.12018

Rhus verniciflua Stokes Extract and Its Flavonoids Protect PC-12 Cells against H2O2-Induced Cytotoxicity 원문보기

Journal of microbiology and biotechnology, v.27 no.6, 2017년, pp.1090 - 1097

Nam, Tae Gyu (Korea Food Research Institute) , Lee, Bong Han (Department of Food Science and Technology, Kyung Hee University) , Choi, Hyo-Kyoung (Korea Food Research Institute) , Mansur, Ahmad Rois (Korea Food Research Institute) , Lee, Sang Gil (Department of Family and Consumer Sciences, North Carolina A&T State University) , Kim, Dae-Ok (Department of Food Science and Technology, Kyung Hee University)

Abstract ▼ AI-Helper

Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated $H_2O_2$-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of $H_2O_2$-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against $H_2O_2$-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against $H_2O_2$-induced neurotoxicity through their antioxidant properties.

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문제 정의

It is well known that brain neuronal cells are vulnerable to ROS, which leads to the development of neurodegenerative diseases [19]; therefore, regulating ROS may be a critical intervention in maintaining cellular functions. In this study, we demonstrate that the RVS EtOAc fraction and its flavonoids effectively inhibit H₂O₂-induced neuronal cell death. The protective effects of the EtOAc fraction and its flavonoids were attributed to their antioxidant capacity, as evidenced by intracellular ROS scavenging and upregulation of SOD activity (Figs.

가설 설정

In this study, we determined the neuroprotective effects of an RVS extract and its flavonoids (butein, sulfuretin, and fustin) in PC-12 cells exposed to H₂O₂. An aqueous methanol extract of RVS was further fractionated to obtain organic solvents and water fractions.

제안 방법

To determine whether H₂O₂-induced cell death is linked to upregulation of intracellular ROS, we measured ROS production using DCFH-DA fluorescence and quantified the intracellular oxidative stress in the presence of the EtOAc fraction, fustin, sulfuretin, and butein (Fig. 3). A decrease in cell viability by more than 10% compared with control cells was considered to be cytotoxic.

대상 데이터

5% B at 27 min, 40% A/60% B at 50 min, 92% A/8% B at 52 min, and 92% A/8% B at 60 min. The detector was set at 280 nm for fustin and 370 nm for sulfuretin and butein. The flow rate was 1 ml/min and the injection volume was 20 µl.

데이터처리

Different letters on the bars indicate significant difference by Duncan’s multiple range test (p < 0.05).

이론/모형

The control group was treated with the same medium without test samples and H₂O₂. Cell viability was measured using MTT reduction assay. Cells were incubated with 0.
4. Effects of the ethyl acetate fraction from Rhus verniciflua Stokes and its flavonoids (fustin, sulfuretin, and butein) on superoxide dismutase (SOD) activity (A) and PC-12 cell viability (B) in the absence and presence of diethyldithiocarbamic acid (DETC) determined using the MTT assay.
The concentration of intracellular ROS was measured using a fluorescent probe (DCFH-DA) assay. After the cells were cultured with the solvent fractions and the three flavonoids identified in the EtOAc fraction for 24 h, they were washed with fresh PBS and incubated with 50 µM DCFH-DA in Hank’s balanced salt solution at 37°C for 30 min.
The total phenolic content of each RVS fraction was determined by using Folin-Ciocalteu’s method [17].

성능/효과

In conclusion, the RVS EtOAc fraction had the highest total phenolic content and antioxidant capacity among all solvent fractions tested. Furthermore, the RVS EtOAc fraction and its flavonoids effectively protected PC-12 cells against H₂O₂-induced cytotoxicity by decreasing caspase-3 activity and reducing H₂O₂-induced oxidative damage, as well as directly scavenging intracellular radicals and enhancing SOD activity.

참고문헌 (29)

Reth M. 2002. Hydrogen peroxide as second messenger in lymphocyte activation. Nat. Immunol. 3: 1129-1134.

상세보기
Ray PD, Huang B-W, Tsuji Y. 2012. Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell. Signal. 24: 981-990.

상세보기
Alzheimer's Association. 2010. 2010 Alzheimer's disease facts and figures. Alzheimers Dement. 6: 158-194.

상세보기
Magliaro BC, Saldanha CJ. 2009. Clozapine protects PC-12 cells from death due to oxidative stress induced by hydrogen peroxide via a cell-type specific mechanism involving inhibition of extracellular signal-regulated kinase phosphorylation. Brain Res. 1283: 14-24.

상세보기
Maher P, Salgado KF, Zivin JA, Lapchak PA. 2007. A novel approach to screening for new neuroprotective compounds for the treatment of stroke. Brain Res. 1173: 117-125.

상세보기
Nichols M, Zhang J, Polster B, Elustondo P, Thirumaran A, Pavlov E, Robertson G. 2015. Synergistic neuroprotection by epicatechin and quercetin: activation of convergent mitochondrial signaling pathways. Neuroscience 308: 75-94.

상세보기
Wolfe KL, Liu RH. 2007. Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements. J. Agric. Food Chem. 55: 8896-8907.

상세보기
Martin MA, Serrano ABG, Ramos S, Pulido MI, Bravo L, Goya L. 2010. Cocoa flavonoids up-regulate antioxidant enzyme activity via the ERK1/2 pathway to protect against oxidative stress-induced apoptosis in HepG2 cells. J. Nutr. Biochem. 21: 196-205.

상세보기
Park S , Kim JA, Choi S, Suh SH. 2010. Superoxide is a potential culprit of caspase-3 dependent endothelial cell death induced by lysophosphatidylcholine. J. Physiol. Pharmacol. 61: 375-381.
Sarker KP, Obara S, Nakata M, Kitajima I, Maruyama I. 2000. Anandamide induces apoptosis of PC-12 cells: involvement of superoxide and caspase-3. FEBS Lett. 472: 39-44.

상세보기
Han X, Zhu S, Wang B, Chen L, Li R, Yao W, et al. 2014. Antioxidant action of 7,8-dihydroxyflavone protects PC12 cells against 6-hydroxydopamine-induced cytotoxicity. Neurochem. Int. 64: 18-23.

상세보기
Jung CH, Jun C-Y, Lee S, Park C-H, Cho K, Ko S-G. 2006. Rhus verniciflua Stokes extract: radical scavenging activities and protective effects on $TH_2O_2$ -induced cytotoxicity in macrophage RAW 264.7 cell lines. Biol. Pharm. Bull. 29: 1603-1607.

상세보기
Jeong EJ, Sung SH, Kim JW, Kim SH, Kim YC. 2008. Rhus verniciflua Stokes attenuates glutamate-induced neurotoxicity in primary cultures of rat cortical cells. Nat. Prod. Sci. 14: 156-160.
Sapkota K, Kim S, Park S-E, Kim S-J. 2011. Detoxified extract of Rhus verniciflua Stokes inhibits rotenone-induced apoptosis in human dopaminergic cells, SH-SY5Y. Cell. Mol. Neurobiol. 31: 213-223.

상세보기
Kwon S-H, Ma S-X, Hwang J-Y, Lee S-Y, Jang C-G. 2015. Involvement of the Nrf2/HO-1 signaling pathway in sulfuretin-induced protection against amyloid beta 25-35 neurotoxicity. Neuroscience 304: 14-28.

상세보기
Park BC, Lee YS, Park H-J, Kwak M-K, Yoo BK, Kim JY, et al. 2007. Protective effects of fustin, a flavonoid from Rhus verniciflua Stokes, on 6-hydroxydopamine-induced neuronal cell death. Exp. Mol. Med. 39: 316-326.

상세보기
Singleton VL, Rossi JA Jr. 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am. J. Enol. Vitic. 16: 144-158.
Kim D-O, Lee KW, Lee HJ, Lee CY. 2002. Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. J. Agric. Food Chem. 50: 3713-3717.

상세보기
Singh RP, Sharad S, Kapur S. 2004. Free radicals and oxidative stress in neurodegenerative diseases: relevance of dietary antioxidants. J. Indian Acad. Clin. Med. 5: 218-225.
Elmore S. 2007. Apoptosis: a review of programmed cell death. Toxicol. Pathol. 35: 495-516.
Jiang B, Liu JH, Bao YM, An LJ. 2003. Hydrogen peroxideinduced apoptosis in pc12 cells and the protective effect of puerarin. Cell Biol. Int. 27: 1025-1031.

상세보기
Olatunji OJ, Feng Y, Olatunji OO, Tang J, Ouyang Z, Su Z. 2016. Cordycepin protects PC12 cells against 6- hydroxydopamine induced neurotoxicity via its antioxidant properties. Biomed. Pharmacother. 81: 7-14.

상세보기
Lee J-C, Lim K-T, Jang Y-S. 2002. Identification of Rhus verniciflua Stokes compounds that exhibit free radical scavenging and anti-apoptotic properties. Biochim. Biophys. Acta 1570: 181-191.

상세보기
Lim K-T, Hu C, Kitts DD. 2001. Antioxidant activity of a Rhus verniciflua Stokes ethanol extract. Food Chem. Toxicol. 39: 229-237.

상세보기
Choi J, An X, Lee BH, Lee JS, Heo HJ, Kim T, et al. 2015. Protective effects of bioactive phenolics from jujube (Ziziphus jujuba) seeds against H2O2-induced oxidative stress in neuronal PC-12 cells. Food Sci. Biotechnol. 24: 2219-2227.

상세보기
Greenlund LJ, Deckwerth TL, Johnson EM. 1995. Superoxide dismutase delays neuronal apoptosis: a role for reactive oxygen species in programmed neuronal death. Neuron 14: 303-315.

상세보기
Shen H-M, Yang C-F, Ding W-X, Liu J, Ong C-N. 2001. Superoxide radical-initiated apoptotic signalling pathway in selenite-treated HepG2 cells: mitochondria serve as the main target. Free Radic. Biol. Med. 30: 9-21.

상세보기
Son Y-O, Lee K-Y, Lee J-C, Jang H-S, Kim J-G, Jeon Y-M, et al. 2005. Selective antiproliferative and apoptotic effects of flavonoids purified from Rhus verniciflua S tokes on norm al versus transformed hepatic cell lines. Toxicol. Lett. 155: 115-125.

상세보기
Cho N, Choi JH, Yang H, Jeong EJ, Lee KY, Kim YC, et al. 2012. Neuroprotective and anti-inflammatory effects of flavonoids isolated from Rhus verniciflua in neuronal HT22 and microglial BV2 cell lines. Food Chem. Toxicol. 50: 1940-1945.

상세보기

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