• 대한임상검사과학회지
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• 제52권4호
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• pp.408-416
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• 2020

본 연구는 치매유발물질의 하나인 염화알루미늄(AlCl3)의 신경독성을 C6 glioma세포를 배양하여 조사하였으며, AlCl3의 독성에 대한 쑥부쟁이[Aster yomena (Kim.), AY] 추출물의 보호효과를 조사하였다. 또한, 세포생존율을 비롯하여 항산화능 분석인 XO 저해능과 SAR-소거능을 조사하였다. 그 결과 AlCl3는 배양 세포에 처리한 농도에 비례적으로 세포생존율을 유의하게 감소시켰다. 이 과정에서 XTT50값이 130.0 μM로 나타나 Borenfreund와 Puerner의 독성판정기준에 의하여 중간독성(mid-cytotoxic)으로 나타났다. 한편, 항산화제인 QU는 AlCl3의 독성으로 손상된 세포생존율을 유의하게 증가시켰다. 또한, AlCl3의 독성에 대한 AY 추출물의 방어효과에 있어서, AY 추출물은 AlCl3만의 처리에 비하여 세포생존율을 유의하게 증가시켰으며, 이와 동시에 XO 저해능과 SAR-소거능과 같은 항산화 효과를 나타냈다. 이상의 결과로부터 AlCl3의 독성에 산화적 손상이 관여하고 있으며, AY 추출물은 항산화 효과에 의하여 AlCl3의 독성을 효과적으로 방어하였다. 따라서, AY 추출물과 같은 천연물질은 질환유발제인 AlCl3와 같이 산화적 손상과 관련이 있는 중금속화합물의 독성 완화나 질환의 치료적 개선을 위한 대체물질개발에 있어 유용한 소재인 것으로 생각된다.

• 대한본초학회지
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• 제30권1호
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• pp.67-75
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• 2015

Objectives : The aim of this study is to examine the neuroprotective effect of wheat bran extract (WBE) against ${\beta}$-amyloid ($A{\beta}$)-induced apoptotic cell death in SH-SY5Y human neuroblastoma cells and memory impairment in triple transgenic animal model's of Alzheimer's disease (3xTg AD mice). Methods : In SH-SY5Y cells, MTT assay and TUNEL staining were conducted to evaluate the protective effect of WBE against $A{\beta}_{25-35}$-induced neurotoxicity and apoptosis. Alterations in mitochondrial transmembrane potential (MMP), expression of proapoptotic Bax and antiapoptotic Bcl-2 proteins, cleavage of PARP, and brain-derived neurotrophic factor (BDNF) levels were analyzed to elucidate the neuroprotective mechanism of WBE. To further investigate the memory enhancing effect of WBE, Morris water maze test was performed in 3xTg AD mice. Results : In SH-SY5Y cells, WBE protected against $A{\beta}_{25-35}$-caused cytotoxicity and apoptosis as shown by the restoration of cell viability in MTT assay and inhibition of DNA fragmentation in TUNEL staining. $A{\beta}_{25-35}$-induced apoptotic signals such as dissipation of MMP, decreased Bcl-2/Bax ratio, and cleavage of PARP were suppressed by WBE. Moreover, WBE up-regulated the protein levels of BDNF, which seemed to be mediated by activation of cAMP response element-binding protein (CREB). In 3xTg AD mice, oral administration of WBE attenuated learning and memory deficit as verified by reduced mean escape latency in water maze test. Conclusions : WBE protects neuronal cells from $A{\beta}_{25-35}$-induced apoptotic cell death and restores learning and memory impairments in 3xTg AD mice. These findings suggest that WBE exhibit neuroprotective potential for the management of AD.

• Preventive Nutrition and Food Science
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• 제16권1호
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• pp.18-23
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• 2011

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by neuronal loss and extracellular senile plaques containing $\beta$-amyloid peptide (A$\beta$). The deposition of the A$\beta$ peptide following proteolytic processing of amyloid precursor protein (APP) by $\beta$-secretase (BACE1) and $\gamma$-secretase is a critical feature in the progression of AD. Among the plant extracts tested, the ethanol extract of Petasites japonicus leaves showed novel protective effect on B103 neuroblastoma cells against neurotoxicity induced by A$\beta$, as well as a strong suppressive effect on BACE1 activity. Ethanol extracts of P. japonicus leaves were sequentially extracted with methylene chloride, ethyl acetate and butanol and evaluated for potential to inhibit BACE1, as well as to suppress A$\beta$-induced neurotoxicity. Exposure to A$\beta$ significantly reduced cell viability and increased apoptotic cell death. However, pretreatment with ethyl acetate fraction of P. japonicus leaves prior to A$\beta$ (50 ${\mu}M$) significantly increased cell viability (p<0.01). In parallel, cell apoptosis triggered by A$\beta$ was also dramatically inhibited by ethyl acetate fraction of P. japonicus leaves. Moreover, the ethyl acetate fraction suppressed caspase-3 activity to the basal level at 30 ppm. Taken together, these results demonstrated that P. japonicus leaves appear to be a useful source for the inhibition and/or prevention of AD by suppression of BACE1 activity and attenuation of A$\beta$ induced neurocytotoxicity.

• 생약학회지
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• 제41권3호
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• pp.166-173
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• 2010

Oxidative stress to proteins, lipids, or DNA is higher in human autopsy tissue and in rodent models of a number of neurodegenerative conditions, including Alzheimer's and Parkinson's disease. On the basis of this information, we established a screening system using N18-RE-105 cells to identify therapeutic agents that can protect cells from glutamate toxicity. During the course of our screening program, we recently isolated the active compound 9-hydroxy-$\alpha$-tocopherone ($\alpha$-TP), which prevents glutamate-induced cell death, from Viola mandshurica. The chemical structure of $\alpha$-TP was identified using spectroscopic methods and by comparison with literature values. Antioxidant activity and protective effects of $\alpha$-TP were evaluated by DPPH radical-scavenging assay, morphological assay, MTT reduction assay, and lactate dehydrogenase (LDH) release assay. These results suggest that $\alpha$-TP could be a new potential chemotherapeutic agent against neuronal diseases.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.69-74
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• 2021

만성 알코올 섭취는 산화적 스트레스 및 치매와 같은 신경퇴행성질환을 발병시킨다. 본 연구는 홍화씨 추출물의 in vitro 항산화 활성과 ethanol로 손상을 유도한 신경교세포에서의 보호 효과를 확인하기 위해 수행되었다. 홍화씨 추출물은 50, 100, 250, 500 ㎍/mL의 농도에서 농도 의존적으로 in vitro에서 1,1-diphenyl-2-prcrylhydrazy (DPPH), hydroxyl radical (·OH), superoxide radical 및 nitric oxide radical 소거능이 증가하였다. 특히 DPPH 및 ·OH radical 소거능 측정 결과, 각각 500 ㎍/mL, 100 ㎍/mL의 농도에서 80% 이상의 radical 소거능을 나타내었다. 홍화씨 추출물의 신경교세포 보호 효과를 확인하기 위해, C6 신경교세포에 500 mM ethanol을 처리하여 산화적 손상을 유도하였다. Ethanol을 처리한 C6 신경교세포는 세포 생존율의 감소 및 reactive oxygen species (ROS) 생성량이 증가하여 세포 손상을 나타내었다. 반면 홍화씨 추출물을 처리하였을 때 세포 생존율 증가 및 ROS 생성 억제를 통해 ethanol로 유도된 신경교세포 손상에 대한 보호 효과를 나타내었다. 본 연구를 통해, 홍화씨는 알코올로 유도된 신경교세포의 산화적 스트레스에 대한 보호 효과가 있는 것으로 사료된다.

• 한국식품과학회지
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• 제50권5호
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• pp.543-548
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• 2018

본 연구는 꾸지뽕 열매 조다당류 분획물(CTP)의 산화방지 활성 및 신경세포 보호 효과에 관하여 알아보기 위하여, 항산화 활성인 라디칼 제거능 및 환원력을 평가한 결과, 꾸지뽕 열매 조다당류의 농도가 증가할수록 산화방지 활성이 유의적으로 증가하는 것으로 나타났다. 또한, 이러한 꾸지뽕 열매 조다당류의 뇌신경세포 보호 효과에 관하여 알아보기 위하여 생쥐의 해마 유래 뇌신경세포에 꾸지뽕 열매 조다당류를 처리한 후 과산화수소로 산화적인 스트레스를 유도하여 세포 독성에 관하여 알아본 결과, 꾸지뽕 열매 조다당류의 처리는 농도 의존적으로 뇌신경 세포의 생존율을 증가시켰으며, 이에 따라 산화방지 효소인 SOD 활성이 증가하고 지방질 과산화 생성물인 MDA level이 감소한 것을 알 수 있었다. 이상의 결과들로 꾸지뽕 열매 조다당류의 산화방지 활성 및 뇌신경세포 보호 효과에 관하여 확인할 수 있었으며, 추후 어떤 메커니즘으로 신경세포를 보호하는지 추가적인 연구가 필요할 것으로 생각한다. 꾸지뽕 열매 조다당류 분획물은 산화방지 및 산화적 스트레스로부터 신경세포 보호효과를 나타내어 퇴행성 신경질환 예방에 유용한 건강기능성 식품 소재가 될 것으로 사료된다.

• 대한한의학회지
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• 제23권2호
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• pp.108-118
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• 2002

Objective : Acupuncture and herbal medicine have been used to prevent and treat cerebrovascular accident, such as stroke, and many studies of acupuncture and moxibustion concerning stroke have been undertaken in humans and various animals. However, the protective effect of the electroacupuncture (EA) of Huan-do (GB30) on the transient forebrain ischemia injury has not been published. Methods : The nenroprotective effects of EA (2 ms, 10 Hz, and 1 - 2 mA) of GB30 on the transient forebrain ischemia injury were investigated by immunohistochemistry of c-Fos-like protein in Sprague-Dawley rats. Results : The transient forebrain ischemia injury resulted in increased expression of c-Fos-like protein (cFL) in the dentate gyms (DG) and CAl for 6 hrs after ischemia, and EA increased significantly expression of cFL in the CAl and DG. For 48 hrs after, there was delayed expression of cFL at the CAl and DG, representing the sign of neuronal cell death, but EA decreased the delayed expression of cFL, significantly. Conclusion : These results suggest that the nenroprotective effects of EA on transient forebrain ischemia injury may be related to excitatory regulation of cFL at the early stage and inhibitory regulation in the long term.

• 대한한의학회지
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• 제21권2호
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• pp.68-78
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• 2000

Objective : This research was performed to investigate protective effects of Sophora subprostrata, against ischemic brain damage after a middle cerebral artery(MCA) occlusion. The effect was estimated using histological test, neurobehavioural test, and biochemical test. Methods : Rats(Sprague-Dawley) were divided into four groups: Sham operated group, MCA occluded group, Sophora subprostrata administrated group after MCA occlusion, and Normal group. The MCA was occluded by intraluminal method. Sophora subprostrata was administrated orally twice(l and 4 hours) after middle cerebral artery occlusion. The neurobeavioural test was performed at 3 hours, 6 hours, 9 hours and 24 hours after the surgery by posture reflex test and swimming behavioural test. All groups were sacrificed at 24 hours after the surgery. The brain tissue was stained with 2% triphenyl tetrazolium chioride(TTC) or 1 % cresyl violet solution, to examine effect of Sophora subprostrata on ischemic brain tissue. The blood samples were obtained from the heart of rats. Tumor necrosis factor-a level was measured from sera using Enzyme-Linked Immunoabsorbent Assay(ELISA). Results : The results showed that (1) Sophora subprostrata reduced infarct size and total infarct volume by 54.8% compared to the control group, (2) that neuronal death, which was shown by decrease in cell number and size, was attenuated significantly in the boundary area of the infarction, (3) that serum $TNF-{\alpha}$ㆍlevel was reduced significantly, and finally, there was significant recovery of motor deficit at 3 hours after MCA occluded by Swimming behavioural test. Conclusions :In conclusion, Sophora subprostrata has protective effects against ischemic brain damage at the early stage of ischemia.

• 대한한의학방제학회지
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• 제25권1호
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• pp.51-68
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• 2017

Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

• Journal of Applied Biological Chemistry
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• 제62권4호
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• pp.327-332
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• 2019

알츠하이머 질환(Alzheimer's disease)은 대표적인 신경퇴행성 질환이며, 뇌 내 amyloid beta (Aβ) 축적에 의한 산화적 스트레스 및 염증반응은 대표적인 AD의 원인으로 알려져 있다. 본 연구에서는 kaempferol (K), kaempferol-3-O-glucoside (KG), quercetin (Q) 및 quercetin-3-β-ᴅ-glucoside (QG)와 같은 4가지 flavonoids의 C6 신경교세포에서 Aβ로 인한 신경독성으로부터의 보호 효능을 살펴보고자 하였다. C6 신경교세포에 Aβ를 처리하였을 때 세포 생존율이 감소한 반면, 4가지 flavonoids 중에서 Q와 QG의 처리 시 세포 생존율 증가를 통해 신경교세포 보호 효과를 확인하였다. 또한, Aβ를 처리한 control군의 경우 reactive oxygen species (ROS) 생성을 유도한 반면, flavonoids의 처리 시 ROS 생성이 감소하였다. 특히 Aβ를 처리한 control군은 133.39%의 ROS 생성을 나타내었으며, 1 μM의 KG와 QG를 각각 처리시 107.44, 113.10%의 수치를 나타내어 ROS 생성 감소를 확인하였다. Flavonoids의 Aβ에 대한 신경교세포 보호 기전을 확인하기 위해 염증 관련 단백질 발현을 측정하였다. Aβ로 신경독성이 유도된 control군은 염증 관련 단백질 발현이 증가하였다. 그러나, flavonoids를 처리한 군의 경우 염증 매개 인자인 inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1β의 발현 감소를 확인하였다. 특히, KG와 QG를 처리한 군은 aglycone 형태인 K와 Q를 처리한 군에 비해 효과적으로 염증 매개 인자 발현을 감소시켰다. 본 연구는 flavonoids의 일종인 K, Q와 그 배당체인 KG, QG의 Aβ로 신경독성이 유도된 신경교세포에서 산화적 스트레스 및 염증반응 조절을 통한 보호 효과를 나타냄을 알 수 있었으며, 이들 생리활성성분은 AD 예방 및 치료 소재로써의 가능성이 있을 것으로 사료된다.