This study aimed to investigate the protective effect of enzymatically modified stevia (EMS) on C2C12 cell-based model of dexamethasone (DEX)-induced muscle atrophy to provide baseline data for utilizing EMS in functional health products. C2C12 cells with DEX-induced muscle atrophy were treated with EMS (10, 50, and 100 μg/mL) for 24 h. C2C12 cells were treated with EMS and DEX to test their effects on cell viability and myotube formation (myotube diameter and fusion index), and analyze the expression of muscle strengthening or degrading protein markers. Schisandra chinensis Extract, a common functional ingredient, was used as a positive control. EMS did not show any cytotoxic effect at all treatment concentrations. Moreover, it exerted protective effects on C2C12 cell-based model of DEX-induced muscle atrophy at all concentrations. In addition, the positive effect of EMS on myotube formation was confirmed based on the measurement and comparison of the fusion index and myotube diameter when compared with myotubes treated with DEX alone. EMS treatment reduced the expression of muscle cell degradation-related proteins Fbx32 and MuRF1, and increased the expression of muscle strengthening and synthesis related proteins SIRT1 and p- Akt/Akt. Thus, EMS is a potential ingredient for developing functional health foods and should be further evaluated in preclinical models.

Methylglyoxal is a highly reactive precursor which forms advanced glycation end products (AGEs). AGEs and methylglyoxal are known to induce various diseases such as diabetes, vascular disorders, Diabetes Mellitus (DM), and neuronal disorders. Juglans regia L is an important food commonly used worldwide, having nutritious components, including phenolic compounds. Since ancient times, Juglans regia L have been differently applied by various countries for health and in diverse diseases, including arthritis, asthma, skin disorders, cancer, and diabetes mellitus. However, the effect of diabetes-induced renal damage against AGEs remains unclear. This study evaluates the anti-glycation and renal protective effects of ethanol extract of Juglans regia L against methylglyoxal-induced renal tubular epithelial cell death. Exposure to methylglyoxal resulted in reduced cell viability in NRK-52E cells, but co-treatment with Juglans regia L extracts significantly increased the cell viability. In addition, we examined the anti-glycation effect of Juglans regia L extracts. Compared to the positive control aminoguanidine and Alagebrium, treatment with Juglans regia L extracts significantly inhibited the formation of AGEs, collagen cross-linking, and breaking collagen cross-linking. Taken together, our results indicate that Juglans regia L is a potential therapeutic agent for regulating diabetic complications by exerting anti-glycation and renal protective activities.

This study was carried out to identify medicinal mushrooms with protective effects against oxidative stress in PC12 neuronal cell line, followed by evaluation of their antioxidant property. Extracts of medicinal mushrooms, including Ganoderma lucidum extract (GLE), antler-shaped Ganoderma lingzhi extract (AGLE), Hericium erinaceus extract (HEE), and Sanghuangporus baumii extract (SBE), were screened for cytotoxicity using MTT assay. None of the extracts up to 10 μg/ml concentration affected cell viability. These extracts were further checked for their protective effect against oxidative stress-induced reactive oxygen species (ROS) production. Exposure to 50 μM H₂O₂ induced ROS generation in PC12 cells, which was inhibited only by treatment with AGLE. In addition, inhibition of H₂O₂-induced ROS generation by AGLE was found to be in a dose-dependent manner (2.5, 5, and 10 μg/ml). Microscopic examination of DCF fluorescence for detection of ROS showed a similar pattern. Further, antioxidant activity of AGLE was determined by ABTS radical cation assay, and its IC50 was found to be 46.90±0.31 μg/ml. Taken together, these results suggest that AGLE may help to alleviate oxidative stress in PC12 neuronal cells.

Amyloid-β protein (Aβ) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of PineXol® on Aβ25-35 induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with 100 μg/mL of PineXol® for 2 h. The cells were exposed to single dose of 30 μM Aβ25-35 for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both Aβ25-35 and PineXol®, compared to the control group. PineXol® induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of Aβ25-35 induced apoptosis. These results suggest that PineXol® may be a good candidate for the prevention of Alzheimer’s disease(AD).

양파 과육과 과피의 생리학적 효과를 평가하기 위하여, in vitro 항산화 활성과 H2O2로 유도된 산화적 스트레스에 대한 PC12 세포보호 효과를 확인하였다. 그 결과, 양파의 과육과 과피 모두 EtOAc 분획물에서 가장 높은 ABTS, DPPH 라디칼 소거활성과 MDA 저해활성을 나타냈다. 양파의 과육과 과피의 EtOAc 분획물을 이용하여, H2O2로 산화적 스트레스를 유발한 PC12세포에서의 ROS 함량과 세포보호효과를 DCF-DA assay, MTT assay 및 LDH assay를 통하여 확인한 결과, 과피가 과육 대비 약 10배 낮은 농도에서 비슷한 보호효과를 나타내었다. 최종적으로 GC-MS를 이용하여 주요 페놀성 화합물인 quercetin 함량과 quercetin 배당체 함량을 분석한 결과, 양파 과육(각각, 4.83, 8.70mg/100g dry weight)과 과피(91.80, 61.81mg/100g dry weight, respectively) 함량을 나타내었다. 이러한 결과들을 종합해볼 때, 양파 과피뿐만 아니라 과육이 나타내는 항산화 효과 및 산화적 스트레스에 대한 신경세포 보호효과는 퇴행성 신경질환과 같은 질병에서 효과적인 소재일 것이라고 사료된다.

본 연구는 식용 버섯의 조리방법에 따른 항산화 생리활성의 평가를 위해 수행되었으며, 산화적 스트레스에 의한 DNA 손상 감소 효과를 통해 조리방법을 달리한 버섯 추출물의 유전독성학적 방호효과를 살펴보았다. Human lymphocyte에 조리방법을 달리한 3가지 버섯(느타리, 팽이, 표고)의 추출물을 처리하고, hydrogen peroxide(H2O2)로 산화적 손상을 준 후, DNA 감소 효과를 Comet assay로 평가한 결과, 모든 시료군에서 산화적 손상에 의한 DNA 손상 감소 효과를 나타냈다. 3가지 버섯 모두 비조리군이 조리군보다 높은 효과를 나타냈는데, 이는 조리과정에 의한 페놀성 화합물의 감소로 인한 것으로 보이며, 조리군 중에서 볶기와 전이 비교적 낮은 DNA 손상 감소 효과를 나타낸 것은 조리 시 첨가되었던 대두유의 가열 산화에 의한 것으로 사료된다. 결론적으로, 조리된 버섯은 생버섯에 비해 산화적 스트레스에 의한 DNA 손상 감소효과가 낮으나, 양성 대조군과 비교하였을 때 손상을 유의적으로 감소시킨 것으로 나타났다. 또한, 본 연구에서 사용한 네 가지 조리법(굽기, 데치기, 볶기, 전) 중 DNA 손상 감소에 효과적인 조리법은 대두유를 사용하지 않은 굽기와 데치기인 것으로 판단된다.

자색고구마 추출물의 항산화 효과와 산화적 스트레스로 유도된 PC12 신경세포에 대한 보호효과에 대하여 연구하 였다. 자색고구마 추출물의 총 페놀함량은 44.25 mg/g, monomeric anthocyanin 함량은 2,394 mg/L로 나타났 다. 자색고구마 추출물의 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylben-zthiazoline- 6-sulfonic acid) (ABTS) radical 소거활성, ferric reducing/antioxidant power (FRAP) 및 환원력은 농도 의존 적으로 항산화 활성이 증가하였다. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazoliumbromide} reduction assay를 이용하여 자색고구마 추출물의 신경세포 보호효과를 측정한 결과, 세포 생존율이 두드러지게 증가하는 것으로 나타났다. 산화적 스트레스는 신경세포막 손상 정도를 증가시키기 때문에 lactate dehydrogenase (LDH) release assay와 neutral red uptake assay를 이용하여 세포막 손상 보호효과를 조사한 결과 자색고구마 추출물 처리구는 대조구에 비하여 산화적 스트레스로 유도된 세포막 손상 보호효과가 농도 의존적으로 나타났다. 따라서 자색고구마 추출물은 천연 항산화 소재 및 알츠하이머성 치매와 같은 신경퇴행성 질환의 예방 소재로서의 활용 가능성이 기대된다.

살충제와 방사선이 자주달개비 수술털 돌연변이에 미치는 복합적 영향을 평가하였다. 포트에서 생육된 Tradescantia 4430 식물체에 코발트 선원으로부터 0.3, 0.5, 1.0 및 2.0 Gy의 감마선을 조사하였다. 살충제는 감마선 조사 24시간 전에 농업용으로 사용되는 파라치온 유제를 살포하였다. CT 실험군과 Pa+γ실험군에 있어서의 돌연변이 빈도가 증가한 고조기간은 방사선 조사 후 7 ~ 11일로 돌연변이 빈도는 방사선량 증가

Background: This study was performed to evaluate the protective effect of Saururus chinensis ethanol extract (SCE) against styrene toxicity in mouse spermatocyte cells [GC-2spd (ts) cell line]. Methods and Results: Cytotoxicity in mouse spermatocyte cells was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Generation of reactive oxygen species (ROS) was determined using 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to quantify the mRNA and protein expression levels, resepectiviely, of stress or apoptosis-related genes including p21, p53, heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), Bax, Bcl-2, and caspase-3. The results of the MTT assay showed that 50 ㎍/㎖ SCE did not affect cell viability. ROS generation in mouse spermatocyte cells increased by treatment with 100 μM styrene, and decreased by co-treatment with SCE. SCE repressed the mRNA expression of stress-related genes, which increased by styrene treatment. In addition, SCE inhibited the apoptosis of mouse spermatocyte cells by ameliorating mRNA and protein levels of apoptotic genes that were altered by styrene treatment. Conclusions: These results suggest that SCE may alleviate styrene toxicity in mouse spermatocyte cells by reducing ROS stress and regulating genes related to styrene toxicity.

본 연구에서는 20종 생약 열수추출물의 AChE 활성, 산화 스트레스로 인한 지질 과산화물 생성 억제능 및 뇌신경 세포 사멸에 대한 보호효과를 비교하였다. AChE의 억제활 성은 산수유, 감초, 당귀 열수추출물에서 우수하였으며, 그 중 산수유가 가장 높은 억제율을 나타내었다. 또한 H2O2/ FeSO4로 산화 스트레스를 유발시켜 생약 열수추출물의 지 질과산화물 생성 억제 활성을 조사한 결과 소엽, 하수오, 계피 및 감초 열수추출물에서 높은 항산화 활성을 보였다. L-Glutamate에 의해 유도된 신경세포 독성에 대한 보호효 과는 감초, 계피, 길경, 박하 열수추출물의 100 μg/mL 농도 에서 관찰되었다. 본 연구를 종합적으로 살펴보았을 때, 감초 열수추출물이 치매 예방 및 개선제로써의 활용가능성 이 가장 뛰어난 것으로 판단된다. 향후 치매예방 효능을 가지는 새로운 화합물 발굴을 위한 기초자료로 활용될 것으 로 사료된다.

Vitis amurensis (VA; Vitaceae) has long been used in oriental herbal medicine. It has been reported that roots and seeds of VA have anti-inflammatory and antioxidant effects. In the present study, the protective effect of ethanol extract from stems and leaves of VA on hydrogen peroxide (H2O2) (100 μm)-induced neuronal cell damage was examined in primary cultured rat cortical neurons. VA (10-100 μg/ml) concentration-dependently inhibited H2O2-induced apoptotic neuronal cell death measured by 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. VA inhibited H2O2-induced elevation of intracellular Ca2+ concentration ([Ca2+]i) and generation of reactive oxygen species (ROS), which were measured by fluorescent dyes. Pretreatment of VA also prevented glutamate release into medium induced by 100 μm H2O2, which was measured by HPLC. These results suggest that VA showed a neuroprotective effect on H2O2-induced neuronal cell death by interfering with H2O2-induced elevation of [Ca2+]i, glutamate release, and ROS generation. This has a significant meaning of finding a new pharmacological activity of stems and leaves of VA in the CNS.

최근 뽕잎과 더불어 기능성 및 천연색소 자원으로서 뽕나무의 열매인 오디가 새로운 작목으로 유망시 되고 있다. 농가의 소득향상은 물론 소비자의 관심과 수요에 부응하기 위한 연구의 일환으로 오디를 수확하여 신경세포 보호활성과 항균활성에 대한 오디 추출물의 효능을 평가함으로써 오디의 기능성 및 이용성을 증대시키고자 하였다. 그 결과를 요약하면 다음과 같다. 1. 신경독성 물질 H2O2 처리에 의해 유도되는 신경세포사에 대하여 오디 추출물 10ug/ml 의 농도 처리시 37~% 의 세포보호 효과를 나타냈으며, Oxygen-glucose deprivation (OGD)에 의하여 유발된 뇌허혈 모델에서는 오디 C3G의 경우 농도 의존형으로 세포괴사를 막았다. 특히 cyanidin의 경우는 10ug/ml 의 농도에서 70~% 이상의 뇌세포보호 효과를 나타냈다. 2. 오디 추출물과 C3G의 항균활성을 검정한 결과, 청일뽕 오디의 MeOH 추출물의 항균 활성이 가장 높았으나, Salmonella typhimurium 의 경우 모든 처리군에서 70~% 이상의 억제 활성을 나타냈다. 모든 처리군에서 50ul 농도보다 100ul 농도에서 균주의 억제 활성이 높게 나타났다.

Sanguisorbae radix (SR) from Sanguisorba officinalis L. (Losaceae) is widely used in Korea and China due to its various pharmacological activity. The present study aims to investigate the effect of the methanol extract of SR on amyloid β Protein(25-35) (Aβ (25-35)), a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. SR, over a concentration range of 10-50 μg/ml, inhibited the Aβ (25-35) (10 μM)-induced neuronal cell death, as assessed by a 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. Pretreatment of SR (50 μg/ml) inhibited 10 μM Aβ (25-35)-induced elevation of cytosolic calcium concentration ([Ca2+]c), which was measured by a fluorescent dye, fluo-4 AM. SR (10 and 50 μg/ml) inhibited glutamate release into medium induced by 10 μM Aβ(25-35), which was measured by HPLC, and generation of reactive oxygen species. These results suggest that SR prevents Aβ (25-35)-induced neuronal cell damage in vitro.