• Title/Summary/Keyword: Free Radical

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Effect of Antioxidants for Porcine Oocytes during In Vitro Maturation, Fertilization and Development (돼지 난포란으로부터 체외수정란의 생산에 있어서 항산화제의 첨가가 배 발달에 미치는 효과)

  • Park H.;Kim J. Y.;Kim J. Y.;Lee J. H.;Park H. D.;Kim J. M.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.245-255
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    • 2004
  • In recent years, an increasing number of studies on pig in vitro maturation(IVM) and in vitro fertilization(IVF) have been separated. the wide range of new technologies, including that in applied molecular genetics, has increased this interest. the production of viable porcine embryos in vitro is a prerequisites for the successful production of transgenic pigs to date. The efficiency of IVM/IVF techniques in the porcine is lower than that obtained in other species such as cattle and mouse. The several problems are generally thought to be the cause of poor results: the low rate of MPN formation derived from inadequate IVM of oocytes, the high incidence of polyspermy after IVF and cell blocking at 4 cell during embryos culture. For there reasons overcoming, many studies have been conducted to improve in vitro embryo-genic competence of oocytes. In the last several years, many maturation culture media have been evaluated and various exogenous factors such as hormones and grows factors have been tested to improve the efficiency of porcine in vitro system. In the study several antioxidants have been examined to improve in vitro fertilization and development of porcine oocytes. In this study, several antioxidants were examined to determine the effects on the development of oocytes to the cleavage, morula and blastocyst stage when added at the maturation(IVM) or in vitro fertilization(IVF) or in vitro culture(IVC) of porcine embryos. Porcine oocytes were matured, fertilized and embryos were cultured in defind conditioned medium in vitro with or without supplementation with the antioxidents of cysteine, catalase and glutathione. 1. Significant improvement of blastocyst rate (27.2% versus 15.4%, p<0.05) were achieved when catalase(500U/$m\ell$) were added to TCM-199 medium and morula rate(72.0% versus 53.9%, p<0.05) were significantly higher when glutathione(1.0mM/$m\ell$) were added to TCM-199 medium than those of control. 2. In mTBM medium for oocytes fertilization, the addition of cysteine, catalase and glutathione had no positive effect on embryonic development. glutathione had no positive effect on embryonic development. In conclusion, this study shows that addition of catalase, gluththione during IVM improved the rate of porcine embryo development.

Effects of ischemic preconditioning, KATP channel on the SOD activation and apoptosis in ischemic reperfused skeletal muscle of rat (허혈양상화와 KATP 통로가 허혈후 재관류된 흰쥐의 골격근육에서 SOD 활성 및 apoptosis에 미치는 영향)

  • Abn, Dong-choon;Paik, Doo-jin;Yang, Hong-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.878-895
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    • 1999
  • Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

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Hydrogen Peroxide Modulates Phospholipase $A_2$ Aactivity and Endogenous Oxidative Stress in the Free Radical Induced Acute Lung Injury (과산화수소에 의한 급성폐손상시 염증성 지질분자의 생성기전에 관한 연구)

  • Bae, Chi-Hoon;Kang, Hyung-Seok;Lee, Sub;Jheon, Sang-Hoon;Ahn, Wook-Wu;Kwon, Oh-Choon
    • Journal of Chest Surgery
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    • v.35 no.5
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    • pp.343-349
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    • 2002
  • background: In an attempt to investigate the role of oxidants in the activation of phospholipase $A_2$(PLA$_2$) and endogenous oxidative stress in the lung. acute inflammatory lung injury was induced by the instillation of hydrogen peroxide into the trachea of Sprague-Dawley rats. Material and Method: To prove the hypothesis thats released oxidants from neutrophils activate the PLA$_2$ retrogradely, activities of PLA$_2$ and lysoplatelet activating factor acetyltransferase(lysoPAF AT) were assayed i hours after instillation of hydrogen peroxide. In addition, to confirm the impairing effects of the activation of PLA$_2$ associated with endogenous oxidative stress, lung weight/body weight ratio(L$\times$10$^{-3}$ B), protein contents(mg/two lungs) in bronchoalveolar lavage(BAL) were measured. As neutrophilic respiratory burst has been known to play a pivotal role in the genesis of endogenous oxidative stress associated with acute inflammatory lung injury, BAL neutrophils counts and level of lung myelperoxidase(MPO) were measured after hydorgen peroxide insult. Morphological and histochemical studies were also performed to identify the effect of the endogenous oxidative stress. Result: Five hours after hydrogen peroxide instillation, lungs showed marked infiltration of neutrophils and increased weight. Protein contents in BAL increased significantly compared to those of normal rats. PLA$_2$ activity was enhanced in the hydrogen peroxide instilled group. Interestingly, the accelerated production of platelet activating factor(PAF) was confirmed by the increased activity of lysoPAF AT in the $H_2O$$_2$ employed lung. Morphologically, light microscopic findings of lungs after instillation of hydrogen peroxide showed atelectasis and infiltration of inflammatory cells, which was thought to be caused by lipid mediators produced by PLA$_2$ activation. In cerium chloride cytochemical electron microscopy, dense deposits of cerrous perhydroxide were identified. In contrast, no deposit of cerrous perhydroxide was found in the normal lung.

Cooking Process for Spinach and Their Effects on Antioxidant and Antimicrobial Activities (조리 과정 중 시금치의 항산화 활성 및 항균 활성의 변화)

  • Park, Cho-Hee;Kim, Kyoung-Hee;Tae, Mi-Hwa;Kim, Na-Young;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.27 no.2
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    • pp.147-155
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    • 2014
  • This study is conducted to investigate the yield of extract, total phenolic compounds, total flavonoid compound contents, free radical scavenging activities (DPPH assay, ABTS assay), reducing power (Oyaizu's assay, FRAP assay) and antimicrobial activities of spinach according to various cooking methods (non-blanched, blanched, seasoned). The yield of non-blanched spinach is 1.64% and the extract yield of blanched spinach is 1.49%, and on the other hand, the yield of seasoned spinach is 6.01%. Total polyphenol contents of seasoned spinach is recorded as $124.31{\pm}1.37mg$ GAE/100 g FW, non-blanched spinach $51.24{\pm}0.27mg$ GAE/100 g FW, and blanched spinach $42.48{\pm}0.53mg$ GAE/100 g FW. From the total flavonoids, seasoned spinach extracts ($15.60{\pm}0.20mg$ CE/100 g FW) showed higher total flavonoid contents than non-blanched. Total antioxidant activities (DPPH assay, ABTS assay, FRAP assay, reducing power) are shown to be in the order of seasoned spinach > non-blanched spinach > blanched spinach. In the antimicrobial activities, non-blanched spinach (5, 10 mg/disc) showed antimicrobial activity against S. enterica and P. aeruginosa. The inhibition zone diameter from extracts of blanched spinach has not been detected. Seasoned spinach indicated antimicrobial activity only against P. aeruginosa (8.15 mm) at 10 mg/disc. If we are to eat a lot of non-blanched spinach, it would cause calculus. Blanching helps to prevent against calculus, since the blanching process can remove various amounts of oxalic acids. The overall results of this study demonstrate that seasoned cooked spinach would be the most efficient way of ingestion to consume antioxidant compounds.

Development of Functional Vinegar by Using Cucumbers (오이를 이용한 기능성 식초 음료 개발)

  • Hong, Sung-Min;Moon, Hyun-Sil;Lee, Ju-Hye;Lee, Hae-In;Jeong, Ji-Hye;Lee, Mi-Kyung;Seo, Kwon-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.7
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    • pp.927-935
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    • 2012
  • This study was performed to develop functional vinegar by using cucumbers through two stages of fermentation. The alcohol content was maximized (7.8%) after 6-days of alcohol fermentation at $25^{\circ}C$ by adjusting the initial sugar concentration to $15^{\circ}Brix$, and vinegar with an acidity of 5.8% was obtained after 12-days of acetic acid fermentation at $30^{\circ}C$. The major sugars in the produced vinegar were glucose and fructose, which were present in concentrations of 3,067.26 and 395.73 mg%, respectively. The major organic acids were acetic acid and succinic acid, which were present in concentrations of 4,410.5 and 841.11 mg%, respectively. The total free amino acid content of the cucumber vinegar was 181.45 ${\mu}g/mL$ and citrulline, valine, aspartic acid, asparagine, and ornithine were the major amino acids. The inorganic components included various alkaline elements, such as K, Ca, and Mg. In addition, experimental methods to assess the DPPH and $ABTS^+$ radical-scavenging ability, reducing power, and ${\beta}$-carotene bleaching activity showed that the cucumber vinegar had strong antioxidant properties. The total polyphenol content, which are the major components responsible for the antioxidant activities of the cucumber vinegar, was 40.14 mg/100 mL. The cucumber vinegar showed significantly higher hepatic aldehyde dehydrogenase activity when compared to the alcoholic control (negative) and the marketing drink (positive), resulting in decreased plasma acetaldehyde concentrations in rats. These results demonstrate that cucumber vinegar possesses antioxidant properties and holds great promise for use in preventing hangovers.

Melanogenesis Inhibitory and Antioxidant Activities of Phellinus baumii Methanol Extract (장수진흙버섯 메탄올 추출물의 멜라닌 생성 저해작용)

  • Lee, J.S.;Shin, D.B.;Lee, S.M.;Kim, S.H.;Lee, T.S.;Jung, D.C.
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.104-111
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    • 2013
  • Phellinus baumii is a medicinal mushroom used in Asian countries for a long period of time. The purpose of this study was to investigate the skin whitening activities of methanol extracts from fruiting bodies of P. baumii. To evaluate the antioxidant activities of the extract, polyphenol and flavonoid contents, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and chelating activity on ferrous ions were studied. For assay of skin whitening activities, tyrosinase and DOPA inhibitory activities, and tyrosinase and melanin synthesis inhibitory activities of B16/F10 melanoma cells treated with the methanol extract were investigated. The total polyphenol content of P. baumii methanol extract was 4.19. DPPH scavenging ability of P. baumii methanol extract was 88.26% in $25{\mu}g/mL$ concentration. We tested tyrosinase inhibitory activity and melanin contents in melanoma cells. The tyrosinase activity was inhibited to 65.17% at the concentration of $125{\mu}g/mL$ and melanin synthesis was inhibited to 57.61% at the concentration of $25{\mu}g/mL$. Overall, the experimental results showed that P. baumii methanol extract had inhibitory activities of tyrosinase and melanin synthesis by dose dependent manner in B16/F10 melanoma cells. Strong ultra-violet absorption spectra in the range of 270~370 nm indicated that ethanol extract of P. baumii could protect the skin from UV. Therefore, P. baumii methanol extract might be used for development of skin whitening, anti-UV and skin care agents.

Antibacterial and Antioxidative Activity of Roasted Coffee and Red Ginseng Mixture Extracts (로스팅 커피와 홍삼 혼합추출물의 항균 및 항산화 효과)

  • Choi, Yu-Hyun;Kim, Sang-Eun;Huh, Jin;Han, Yeong-Hwan;Lee, Moon-Jo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.3
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    • pp.320-326
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    • 2012
  • This study was conducted to investigate the antibacterial and antioxidative activities of water and ethanol extracts from a mixture of roasted coffee and red ginseng. The antibacterial effects of each extract were determined by the classical paper disc method. A water extract of mixture samples inhibited growth of all strains, but antibacterial effects were mostly weakened. Ethanol extracts showed stronger antibacterial effects than water extracts in all strains except Gram negative Escherichia coli and the fungi strain Candida albicans. Also, the antibacterial effect of the Bacillus cereus strain appeared in all samples, and the ES2 sample formed a clear zone of 19 and 20 mm against Pseudomonas aeruginosa and S. Typhimurium respectively (MIC=0.25 and 0.125 mg/mL). Determinations of free radical elimination for the different mixture extracts using 1,1-diphenyl-2-picrylhydrazyl (DPPH) were compared with ascorbic acid and butylated hyderoxytoluene as positive controls. The water and ethanol extracts of mixture samples (100 ${\mu}g/mL$) showed 55.38~60.01% and 59.37~70.50% DPPH scavenging activities, respectively. DPPH scavenging activities of all mixture samples were slightly higher than roasted coffee and red ginseng samples. However, DPPH scavenging activity decreased when red ginseng extract composed more than 70% of the total extract. The total polyphenol in the mixture samples measured by the Folin-Denis method revealed the highest level of polyphenol content in ethanol extract of sample 3, whereas polyphenol content differed with different mixture ratios, ranging from 105.16~119.79 mg/g in ethanol extract. In the water extract, the polyphenol content was greatest with water extract of sample 1, whereas in other samples the content varied from 93.75~109.18 mg/g.

Effect of PE Film Mulching and Irrigation Method on the Growth, Yield and Antioxidant Activity for Potatoes Grown in Winter Season at Saemangeum Reclaimed Land (새만금 간척지에서 감자 겨울재배시 비닐멀칭 및 관수방법이 생육, 수량 및 괴경의 항산화 활성에 미치는 영향)

  • Choi, Weon-Young;Cho, Kwang-Min;Kim, Sun;Jeong, Jae-Hyeok;Lee, Su-Hwan;Lee, Kyeong-Bo;Lee, Geon-Hwi;Park, Ki-Hun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.63-69
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    • 2015
  • This study was conducted to measure the effect of PE film mulching and irrigation methods on the growth, yield and antioxidant activity of potatoes tubers, in order to examine the possibility of cultivating potatoes in winter season in vinyl greenhouse on the reclaimed tidal land with weak ground inside the sea wall currently completed. The test was conducted on the sandy loam soil (Munpo series), and its salt concentration was 0.42% at the time of planting. The emergence speed per kind of PE film mulching was in the order of black > coloration > transparent > green color, with the black color showing the fastest speed. The temperature change during a day per kind of PE film mulching was in the order of transparent > coloration > green > black color. As for the salt concentration in the soil for each different way of water management, the salt concentration in the treatment of drip irrigation with 1 week interval was lower than that drip irrigation with 2 weeks interval. As for the growth of above-aerial part, plant length was higher, number of tiller and leaves were more and dry weight of above-aerial part was larger in the treatment of drip irrigaton with 1 week interval than drip irrigation with 2 weeks interval. As for the yield of potatoes depending on each way of water management, the yield in the treatment of drip irrigation with 1 week interval was more than drip irrigation with 2 weeks interval. The yield for each different kind of PE film mulching in the weekly drip-irrigation management section was in the order of transparent ${\geq}$ black ${\geq}$ coloration ${\geq}$ green color. In both of total phenol content and DPPH free radical activity experiments, the content and activity were higher with pear color vinyl treatment.

Effects of Scutellaria radix Extract on Osteoblast Differentiation and Osteoclast Formation (황금 추출물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Shin, Jeong-Min;Park, Chan-Kyung;Shin, Eun-Ju;Jo, Tae-Hyung;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.674-679
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    • 2008
  • Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.

Nutritional composition, antioxidant capacity, and brain neuronal cell protective effect of cultivars of dried persimmon (Diospyros kaki) (품종별 곶감(Diospyros kaki)의 영양성분 분석, 산화방지 효과 및 뇌 신경세포 보호효과)

  • Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Park, Sang Hyun;Park, Su Bin;Yoo, Seul Ki;Han, Hye Ju;Lee, Su-Gwang;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.225-237
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    • 2018
  • This study was conducted to compare nutritional analysis and neuroprotective effect of 5 cultivars of Diospyros kaki (Dungsi, Godongsi, Gojongsi, Gabjubaekmok, and Bansi). In nutritional analysis, three free sugars: sucrose, glucose, and fructose, and six fatty acids: tartaric acid, hexadecanoic acid, palmitic acid, oleic acid, octadecenamide, and octadecane, were detected. Potassium and phosphorus levels were the highest in inorganic component analysis, and glutamic acid and aspartic acid were the highest contents in amino acid analysis. Vitamin C was detected in all cultivars. Total phenolic content was the highest in Dungsi. Antioxidant activities such as ABTS (3-ethylbenzothiazoline-6-sulfonic acid), DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activities, FRAP (ferric reducing/antioxidant power), and MDA (malondialdehyde) inhibitory effect were the highest in Gabjubaekmok. Acetylcholinesterase inhibitory activity, cell viability, intracellular reactive oxygen species (ROS) accumulation, and lactate dehydrogenase (LDH) release were measured to confirm the neuroprotective effect in MC-IXC cells. Gabjubaekmok showed significant acetylcholinesterase (AChE) inhibition and neuroprotection.