Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2 , Gsr , HO-1, and catalase , were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2 , HO-1 , and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

최근 기능성과 친환경 화장품에 대한 관심이 증가하고 있으며, 이에 따라 안전하면서 효능이 우수한 식물 추출물을 활용한 소재 개발이 이루어지고 있는 실정이다. 따라서 본 연구에서도 주로 건강 기능성 소재로써 다양한 효능이 있는 것으로 알려진 그라비올라 추출물이 기능성 화장품 소재로써의 가 능성을 확인하고자 하였다. 그라비올라 추출물의 항산화 활성을 확인하고자 총 폴리페놀과 총 플라보노 이드 함량, DPPH radical 소거 활성을 측정하였고, HDF 세포에서의 세포 독성을 확인한 후 적정 농도 에서 HDF 세포에 과산화수소(H2O2)를 처리하여 산화적 스트레스에 대한 ROS 활성 억제 효과와 세포 보호 효과를 측정하였다. 본 실험 결과, 그라비올라 추출물은 항산화 지표가 되는 총 폴리페놀과 플라보 노이드의 100g당 26.6 mg(CA)/100g, 14.3 mg(CA)/100g의 높은 함량을 확인하였으며, 높은 radical 소 거 활성을 확인하였다. HDF 세포에 대한 세포 생존율을 측정한 결과, 모든 농도에서 유의한 세포 독성 이 나타나지 않았으며, 추후 100 μg/mL 농도에서 실험하였다. H2O2로 유도된 HDF 세포에 ROS 활성 억제를 측정한 결과, 농도 의존적인 ROS 활성 억제 효과를 확인하였고, H2O2를 4 시간, 24 시간, 48 시간 동안 처리 후 그라비올라 추출물의 세포 보호 효과를 측정한 결과, 25 μg/mL 농도에서 24시간까 지 89.92%의 높은 세포 보호 효과를 확인하였다. 이와 같은 결과를 통하여 그라비올라 추출물은 항산 화 활성이 우수하고, HDF 세포에 대한 독성이 거의 없으며, H2O2에 의해 발생하는 활성산소에 대한 효과적인 활성 억제 효과와 세포 보호 효과가 우수한 것으로 확인됨에 따라 항산화 및 세포 보호 효과 를 가진 다양한 기능성 소재로서의 가능성을 확인하였다.

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.

Inflammatory bowel disease (IBD) is a group of chronic disorders of unknown etiology characterized by inflammation of the gastrointestinal tract. Recent data showed that the development of IBD is associated with the interplay of genetic, bacterial, and environmental factors and dysregulation of the intestinal immune system. We investigated how the gut cells were repaired after injury in Drosophila melanogaster. In this study we made D. melanogaster intestine damage model by oral feeding with variety IBD inducer such as pathogenic bacteria Serratia marcescens, Dextran Sulfate Sodium (DSS) and bleomycin, because its function is very similar with human, even though D. melanogaster has relatively simple organism. We repeated oral feeding with variety IBD inducer and got the survival rate and 50% lethal dose (LD50). After feeding with IBD inducer, we investigated the change of the intestinal stem cells, innate immune-related gene expression, and apoptosis in D. melanogaster gut. We examined the Delta, stem cell marker, staining image in the gut after feeding with DSS and S. marcescens with LD50 concentration. The Delta positive cells greatly increased in gut cells damaged by DSS or S. marcescens. This result supports the idea that intestinal gut stem cells are increased after gut cell damage and play very important role in damaged cell repair. Expression level of antimicrobial peptides was dramatically up-regulation after gut damage. As a result of TUNEL (terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling) assay, we confirmed that cell death by apoptosis was very increased in DSS feeding flies. Accordingly, we suggest that D. melanogaster is a proper IBD model organism to study how intestine damage can be repaired.

The present study reports the protective properties of a total methanol extract of B. platyphylla var. japonica against ultraviolet (UV)-C irradiation. Pretreatment of Chinese hamster fibroblast (V79-4) cells with a total methanol extract significantly increased cell survival following 300J/m² of UV-C irradiation. The total methanol extract was further fractionated into 5 fractions: n-hexane, dichloromethane, ethylacetate, n-butanol and water fractions. Among these fractions, B. platyphylla var. japonica ethylacetate, butanol and water fractions showed significant protective effects against the cellular damage induced by UV-C irradiation. In order to elucidate the mechanism underlying this protective effect, DPPH (Editor note: abbreviations should be spelled out at first use.) radical scavenging and lipid peroxidation inhibitory activity were measured. Significant radical scavenging and lipid peroxidation inhibitory activities were observed for the ethylacetate fraction. In summary, the present data demonstrate that an extract of B. platyphylla var. japonica has a significant protective effect against UV-C irradiation. The underlying mechanism of this protective effect may involve radical scavenging and inhibition of lipid peroxidation by the B. platyphylla var. japonica extract.

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

Potentially lethal damage repair (PLDR) in HFL-I was investigated by delayed plating experiments. The surviving fraction data were fitted to the linear Quadratic equation (LogSn=-nγ(αd+βd2) where γ=1 for immediate plating). And a repair factor γ was developed to compare survival for immediate and delayed plating . When we only took into account the repair factor of PLDR γ which was derived from the delay assay, the cell survival response th fractionated carbon ion irradiation was not fully matched. This gap suggested that consideration of another repair process is necessary. So this suggests that the various repair process plays an important role in the fractionated irradiations.

Dieldrin, one of the organochlorine pesticides (OCPs), induced the damages in neuroblastoma cells and DNA damages in lymphocytes. The ethanol extracts of A. sessiliflorus leaves were examined for the suppressive effects on the dieldrin-induced cell damages. Moreover, the extract was used to test whether it might inhibit the oxidative DNA damage of lymphocytes using Comet assay. The cell and DNA damage by dieldrin were suppressed in vitro upon treating A. sessiliflorus extract. This result suggests that A. sessiliflorus extract might be useful to reduce dieldrin toxicity.

본 연구는 췌장베타세포인 HIT-T15 세포를 이용하여 매실주정추출물(PME)의 alloxan에 의한 산화스트레스로부터의 세포보호, 인슐린 분비능 및 항산화 효소 활성을 평가하였다. PME는 alloxan에 의해 유발된 산화스트레스로부터 세포를 보호하여 세포생존율을 증가시켰다. PME는 세포막 손상지표인 LDH 방출을 억제하였고 NAD+/NADH ratio를 유의적으로 증가시켜 세포사멸이 억제되어짐을 확인하였다. 또한 alloxan 단독처리군에 비해 250 μg/ml PME 처리군에서 인슐린 분비량이 유의성 있게 증가하였다. Alloxan과 PME를 동시에 처리하여 HIT-T15세포의 항산화효소 활성을 측정했을 때 산화스트레스에 의해 감소되었던 항산화효소 활성이 PME에 의해 보호되는 효과를 확인하였다. 이상의 연구결과로부터 PME는 세포괴사 및 DNA fragmentation을 억제하고 세포내 항산화효소 활성을 증가시켜 alloxan에 의해 유발된 산화스트레스로부터 췌장베타세포를 보호하고 이에 따른 인슐린 분비능 조절 효과가 있는 것으로 생각된다.

Ethanol treatment during the brain growth spurt period has been known to induce the death of Purkinje cells. The underlying molecular mechanisms and the role of reactive oxygen species (ROS) in triggering ethanol-induced Purkinje cell death are, however, largely unresolved. We undertook TUNEL staining, western blotting assay and immunohistochemistry for the cleaved forms of caspase-3 and -9, with calbindin D28K double immunostaining to identify apoptotic Purkinje cells. The possibility of ROS-induced Purkinje cell death was immunohistochemically determined by using anti-8-hydroxy-2'deoxyguanosine (8-OHdG), a specific cellular marker for oxidative damage. The results show that Purkinje cell death of PD 5 rat cerebellum following ethanol administration is mediated by the activation of caspase-3 and -9. However, unexpectedly, TUNEL staining did not reveal any positive Purkinje cells while there were some TUNEL-positive cells in the internal and external granular layer. 8-OHdG was detected in the Purkinje cell layers at 8 h, peaked at 12-24 h, but not at 30 h post-ethanol treatment. No 8-0HdG immunoreactive cells were detected in the internal and external granular layer. The lobule specific 8-OHdG staining patterns following ethanol exposure are consistent with that of ethanol-induced Purkinje cell loss. Thus, we suggest that ethanol-induced Purkinje cell death may not occur by the classical apoptotic pathway and oxidative damage is involved in ethanol-induced Purkinje cell death in the developing cerebellum.

본 연구에서는 열수 팥 추출물이 hydroxyl 라디칼에 의해 유도되는 산화적 스트레스에 미치는 영향을 알아보기 위하여 항산화활성과 DNA 및 세포의 산화적 손상 억제 효과를 조사하였다. 팥 열수 추출물의 DPPH 라디칼과 hydroxyl 라디칼의 제거능은 다소 낮았으나, Fe2+-chelating과 과산화수소 제거효과는 높게 나타나 활성산소의 생성을 억제하는 데 효과적인 것으로 확인되었다. 또한 팥 열수 추출물의 in vitro DNA cleavage, DNA migration 및 H2AX의 인산화비 억제활성은 높은 활성을 보여주고 있어 라디칼에 의한 DNA 손상 억제에 효과적으로 작용하였다. 또한 지질과산화와 p21의 발현율을 통해 세포의 산화적 손상에 미치는 영향을 살펴보면 지질과산화 억제능과 p21의 발현율에 매우 효과적으로 작용하고 있어 라디칼에 의한 산화적 스트레스로부터 세포를 보호할 것으로 생각된다.

산화적 스트레스로부터 참다래 과실 추출물의 신경세포 보호효과에 미치는 영향을 알아보기 위하여 신경세포주인 PC12 세포를 이용하여 참다래 과실추출물의 전처리가 산화적 손상으로부터 유발되는 신경세포사멸을 억제할 수 있는지 조사하였다. t-BHP에 의해 유도된 신경세포손상으로부터 세포사멸을 억제하여 세포생존도를 증가시켰으며 세포사멸로부터 형성되는 핵의 농축현상과 단편화가 현저히 감소함을 확인 할 수 있었다. 그리고 Bcl-2 단백의 발현 증가, Bax 단백의 발현 감소, caspase-3의 활성, PARP 분해 단백(85KDa)감소, ERK, p38 활성을 감소시켰다. 따라서 참다래 과실의 추출물은 신경세포증식효과를 통해 신경세포손상으로부터 유발되는 다양한 퇴행성 뇌질환의 예방에 도움이 될 것으로 나타났다.

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.

Dried leaves from Cedrela sinensis A. Juss. (CS), have been observed to possess various pharmacological activity and contain various antioxidant constituents. The protective effect of ethanol extract of CS on hydrogen peroxide (H2O2)-induced neurotoxicity was examined using primary cultured rat cortical neurons in the present study. Exposure of cultured neurons to 100 μM H2O2 caused a significant neuronal death as assessed by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. The addition of CS, over a concentration range of 10 to 50μg/ml, concentration-dependently prevented the H2O2-induced neuronal apoptotic death. CS (50μg/ml) significantly inhibited H2O2-induced elevation of the cytosolic Ca2+ concentration ([Ca2+]c), which was measured by a fluorescent dye, Fluo-4 AM. CS (30 and 50μg/ml) inhibited glutamate release and generation of reactive oxygen species (ROS) induced by 100μM H2O2. These results suggest that CS may mitigate the H2O2-induced neurotoxiciy by interfering with the increase of [Ca2+]c, and then inhibiting glutamate release and generation of ROS in cultured neurons.

Endocrine disrupting compounds (EDC's) are chemicals that either mimic endogenous hormones interfering with pharmacokinetics or act by other mechanisms. Some endocrine disrupters were reported to be chemical substances that cause apoptosis in cells. A number of reports have indicated that 1,3-DCP, one of the EDC's may act as an endocrine disrupter and also has possible carcinogenic effects. 1,3-DCP, present in commercial protein hydrolysates used for human nutrition, are genotoxic and 1,3-dichloro-2-propanol induced tumors in rats. In the present study, it was investigated whether 1,3-DCP induces ROS generation and apotosis in A549 adenocarcinoma cells. Here we show that 1,3-DCP inhibits the growth of lung cancer cell lines and generates reactive oxygen species (ROS), a major cause of DNA damage and genetic instability. It was investigated that 1,3-DCP increases G1 phase cells after 12 hours, thereafter abruptly draws A549 cells to G0 state after 24 hours by flow cytometric analysis. 1,3-DCP induces p53 and p21Cip1/WAF1 activation time- and dose-dependently by 24 hours, while the level p21Cip1/WAF1 was decreased after 48 hours. These results suggest that 1,3-DCP, an EDC's generates ROS and regulates genes involved with cell cycle and apoptosis.

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.

Caulis Bambusae in Taenia 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 Caulis Bambusae in Taenia (CB) from Phyllostachys nigra Munro var. henonis Stapf (Gramineae) on amyloid β protein (25-35) (Aβ (25-35)), a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. CB, over a concentration range of 10-50μg/μl, inhibited the Aβ (25-35) (10 μM)-induced neuronal cell death, as assessed by a 3-[4,5-dimethyIthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. CB (50 μg/μl) inhibited glutamate release into medium induced by 10 μM Aβ, (25-35) which was measured by HPLC. Pretreatment of CB (50 μg/μl) inhibited 10μM Aβ (25-35)-induced elevation of cytosolic calcium concentration ([Ca2+]c), which was measured by a fluorescent dye, fluo-4 AM, and generation of reactive oxygen species. These results suggest that CB prevents Aβ (25-35)-induced neuronal ell damage in vitro.