• 한국응용과학기술학회지
• /
• 제37권3호
• /
• pp.418-428
• /
• 2020

본 연구는 알츠하이머(Alzheimer's disease: AD) 형질전환 생쥐를 대상으로 저항성 운동(resistance exercise: RE)이 해마의 베타 아밀로이드(β-amyloid: Aβ) 단백질 대사, 신경세포사멸 및 인지기능에 미치는 영향을 확인하는데 목적이 있다. AD 비 형질전환 생쥐(non-transgenic: non-tg, n=14)와 형질전환 생쥐(transgenic: Tg, n=14)를 무선 배정하여 비 형질전환 생쥐 대조군 (non-tg-control: NTC, n=7), 비 형질전환 생쥐 저항성 운동군(non-tg-RE: NTRE, n=7), 형질전환 대조군(tg-control: TC, n=7) 및 형질전환 저항성 운동군(tg-RE: TRE, n=7)으로 구분하였다. RE는 특수 제작한 사다리 저항성 운동 기구를 사용하여 점진적으로 set 수를 증가시켜 총 8주간 실시하였다. 운동 후 인지기능 능력을 평가하기 위한 수중미로검사와 Aβ 단백질 대사, 신경세포사멸 지표 및 SIRT1/PGC-1α 단백질 발현 수준을 확인하였다. 수중미로검사 결과 거리와 시간 모두 TC 집단에서 유의하게 증가 되었지만 RE를 실시한 TRE 집단에서 거리와 시간이 감소 되어 인지능력이 개선된 것으로 확인되었다. 또한, TC 집단에서 증가된 Aβ 단백질 발현은 RE를 통해 감소하는 것으로 나타났다. 신경세포사멸 관련 단백질인 Bcl-2/Bax ratio는 TC 집단에서 유의하게 감소되어 신경세포사멸이 증가 된 것으로 나타났지만 RE는 Bcl-2/Bax ratio을 증가시켜 신경세포사멸을 감소시킨 것으로 확인되었다. TC 집단에서 증가된 BACE1 및 ROCK1과 감소된 ADAM10과 RARβ 단백질 발현은 RE를 통해 감소되거나 증가 된 것으로 나타났고, SIRT1/PGC-1α 단백질 발현은 TC 집단에서 감소 되었지만 RE를 통해 증가 된 것으로 나타났다. 따라서 8주간의 RE는 AD의 병리학적 특징인 Aβ 단백질 발현을 감소시키고 관련 생성 기전들을 조절하여(SIRT1/PGC-1α 기전 활성, 아밀로이드 생성기전 억제, 비-아밀로이드 생성기전 활성) 신경세포사멸 억제시키고 결과적으로 인지기능을 개선 시킬 수 있는 효과적인 운동 방법이라고 생각된다.

• BMB Reports
• /
• 제44권2호
• /
• pp.135-139
• /
• 2011

Chronic alcohol consumption contributes to numerous diseases, including cancers, cardiovascular diseases, and liver cirrhosis. Epidemiological studies have shown that excessive alcohol consumption is a risk factor for dementia. Along this line, Alzheimer's disease (AD) is the most common form of dementia and is caused by the accumulation of amyloid-$\beta$ ($A{\beta}$ plaques in neurons. In this study, we hypothesized that chronic ethanol consumption is associated with pathological processing of APP in AD. To investigate the relationship between chronic alcohol consumption and $A{\beta}$ production, brain samples from rats fed an alcohol liquid diet for 5 weeks were analyzed. We show that the expression levels of APP, BACE1, and immature nicastrin were increased in the cerebellum, hippocampus, and striatum of the alcohol-fed group compared to the control group. Total nicastrin and PS1 levels were induced in the hippocampus of alcohol-fed rats. These data suggest that the altered expression of APP and $A{\beta}$-producing enzymes possibly contributes to the chronic alcohol consumption-mediated pathogenesis of AD.

• BMB Reports
• /
• 제43권10호
• /
• pp.704-709
• /
• 2010

The Swedish mutation (K595N/M596L) of amyloid precursor protein (APP-swe) has been known to increase abnormal cleavage of cellular APP by Beta-secretase (BACE), which causes tau protein hyperphosphorylation and early-onset Alzheimer's disease (AD). Here, we analyzed the effect of APP-swe in global gene expression using deep transcriptome sequencing technique. We found 283 genes were down-regulated and 348 genes were up-regulated in APP-swe expressing H4-swe cells compared to H4 wild-type cells from a total of approximately 74 million reads of 38 base pairs from each transcriptome. Two independent mechanisms such as kinase and phosphatase signaling cascades leading hyperphosphorylation of tau protein were regulated by the expression of APP-swe. Expressions of catalytic subunit as well as several regulatory subunits of protein phosphatases 2A were decreased. In contrast, expressions of tau-phosphorylating glycogen synthase kinase $3\beta$(GSK-3$\beta$), cyclin dependent kinase 5 (CDK5), and cAMP-dependent protein kinase A (PKA) catalytic subunit were increased. Moreover, the expression of AD-related Aquaporin 1 and presenilin 2 expression was regulated by APP-swe. Taken together, we propose that the expression of APP-swe modulates global gene expression directed to AD pathogenesis.

• Molecules and Cells
• /
• 제27권6호
• /
• pp.651-656
• /
• 2009

The formation of ${\beta}$-amyloid peptide ($A{\beta}$) is initiated from cleavage of amyloid precursor protein (APP) by a family of protease, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-secretase. Sub W, a substrate peptide, consists of 10 amino acids, which are adjacent to the ${\beta}$-cleavage site of wild-type APP, and Sub M is Swedish mutant with double mutations on the left side of the ${\beta}$-cleavage site of APP. Sub W is a normal product of the metabolism of APP in the secretary pathway. Sub M is known to increase the efficiency of ${\beta}$-secretase activity, resulting in a more specific binding model compared to Sub W. Three-dimensional structures of Sub W and Sub M were studied by CD and NMR spectroscopy in water solution. On the basis of these structures, interaction models of ${\beta}$-secretase and substrate peptides were determined by molecular dynamics simulation. Four hydrogen bonds and one water-mediated interaction were formed in the docking models. In particular, the hydrogen bonding network of Sub M-BACE formed spread over the broad region of the active site of ${\beta}$-secretase (P5-P3'), and the side chain of P2- Asn formed a hydrogen bond specifically with the side chain of Arg235. These are more favorable to the cleavage of Sub M by ${\beta}$-secretase than Sub W. The two substrate peptides showed different tendency to bind to ${\beta}$-secretase and this information may useful for drug development to treat and prevent Alzheimer's disease.

• Biomolecules & Therapeutics
• /
• 제28권3호
• /
• pp.259-266
• /
• 2020

The present research work primarily investigated whether spinosin has the potential of improving the pathogenesis of Alzheimer's disease (AD) driven by β-amyloid (Aβ) overproduction through impacting the procession of amyloid precursor protein (APP). Wild type mouse Neuro-2a cells (N2a/WT) and N2a stably expressing human APP695 (N2a/APP695) cells were treated with spinosin for 24 h. The levels of APP protein and secreted enzymes closely related to APP procession were examined by western blot analysis. Oxidative stress related proteins, such as nuclear factor-erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) were detected by immunofluorescence assay and western blot analysis, respectively. The intracellular reactive oxygen species (ROS) level was analyzed by flow cytometry, the levels of Aβ_1-42 were determined by ELISA kit, and Thioflavin T (ThT) assay was used to detect the effect of spinosin on Aβ_1-42 aggregation. The results showed that ROS induced the expression of ADAM10 and reduced the expression of BACE1, while spinosin inhibited ROS production by activating Nrf2 and up-regulating the expression of HO-1. Additionally, spinosin reduced Aβ_1-42 production by impacting the procession of APP. In addition, spinosin inhibited the aggregation of Aβ_1-42. In conclusion, spinosin reduced Aβ_1-42 production by activating the Nrf2/HO-1 pathway in N2a/WT and N2a/APP695 cells. Therefore, spinosin is expected to be a promising treatment of AD.

• Journal of Ginseng Research
• /
• 제47권3호
• /
• pp.458-468
• /
• 2023

Background: As a complication of Type II Diabetes Mellitus (T2DM), the etiology, pathogenesis, and treatment of cognitive dysfunction are still undefined. Recent studies demonstrated that Ginsenoside Rg1 (Rg1) has promising neuroprotective properties, but the effect and mechanism in diabetes-associated cognitive dysfunction (DACD) deserve further investigation. Methods: After establishing the T2DM model with a high-fat diet and STZ intraperitoneal injection, Rg1 was given for 8 weeks. The behavior alterations and neuronal lesions were judged using the open field test (OFT) and Morris water maze (MWM), as well as HE and Nissl staining. The protein or mRNA changes of NOX2, p-PLC, TRPC6, CN, NFAT1, APP, BACE1, NCSTN, and Ab1-42 were investigated by immunoblot, immunofluorescence or qPCR. Commercial kits were used to evaluate the levels of IP3, DAG, and calcium ion (Ca²⁺) in brain tissues. Results: Rg1 therapy improved memory impairment and neuronal injury, decreased ROS, IP3, and DAG levels to revert Ca²⁺ overload, downregulated the expressions of p-PLC, TRPC6, CN, and NFAT1 nuclear translocation, and alleviated Aβ deposition in T2DM mice. In addition, Rg1 therapy elevated the expression of PSD95 and SYN in T2DM mice, which in turn improved synaptic dysfunction. Conclusions: Rg1 therapy may improve neuronal injury and DACD via mediating PLC-CN-NFAT1 signal pathway to reduce Aβ generation in T2DM mice.

• 대한한의학방제학회지
• /
• 제25권1호
• /
• pp.69-87
• /
• 2017

Objectives : Cheongnoemyeongsin-hwan (CNMSH) is a herb medicine to treat cognitive impairment. This study was investigated the effects of CNMSH on learning and memory impairment induced by cerebral hypoperfusion. Cerebral hypoperfusion was produced chronically by permanent bilateral common carotid artery occlusion (BCCAO) in rats. Methods : CNMSH was administered orally once a day (250 mg/kg) for 28 days starting at 4th week after the BCCAO. The acquisition of learning and the retention of memory were tested on 9th week after the BCCAO using the Morris water maze. In addition, effect of CNMSH on neuronal apoptosis and ${\beta}-amyloid$ accumulation in the hippocmapus was evaluated with immunohistochemistry and Western blotting. Results : 1. CNMSH and ChAL significantly shortened the escape latencies on the 2nd day of acquisition training trials. 2. ChAL significantly prolonged the swimming time spent in the target and peri-target zones and CNMSH also significantly prolonged the swimming time spent in the peri-target zone. 3. CNMSH and ChAL significantly increased the number of target heading in the retention test. 4. ChAL significantly shortened the time of the 1st target heading in the retention test, but CNMSH insignificantly shortened the time of that. 5. CNMSH and ChAL significantly increased the memory score in the retention test. 6. CNMSH and ChAL significantly attenuated the reduction of CA1 neurons, but insignificantly attenuated the reduction of CA1 thickness. 7. CNMSH and ChAL significantly attenuated the up-regulation of Bax expression in the CA1 of hippocampus. 8. CNMSH and ChAL significantly attenuated the up-regulation of cascapse-3 expression in the CA1 of hippocampus. 9. CNMSH and ChAL significantly attenuated the ${\beta}-amyloid$ accumulation in the CA1 of hippocampus. 10. CNMSH and ChAL significantly attenuated the up-regulation of APP expression in the CA1 of hippocampus. 11. CNMSH and ChAL significantly attenuated the up-regulation of BACE-1 expression in the CA1 of hippocampus. Conclusions : The results show that CNMSH attenuates neuronal apoptosis and ${\beta}-amyloid$ accumulation in the hippocampus and alleviates the impairment of learning and memory produced by chronic cerebral hypoperfusion. These results suggest that CNMSH may be a beneficial medicinal herb to treat cognitive impairment associated with neurodegenerative diseases.

• 대한한의학회지
• /
• 제27권2호
• /
• pp.122-133
• /
• 2006

Objectives : The amyloid $\beta$-protein ($A\beta$) is the principal component of the senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors including antioxidants and proteoglycans modify $A{\beta}toxicity$. In this study, we have investigated the protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A\beta$-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. Methods : For this study, we used MTT reduction assay for detection of protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used cell-based $\beta$-secretase assay system to investigate the inhibitory effect of water- and organic solvent-extracts of Hemerocallis fulva root on $\beta$-secretase activity. Results : We previously reported that methanol extracts of Hemerocallis fulva root strongly attenuated cytotoxicity induced by the three $A\beta$ fragments ($A{\beta}_{25-35},\;A{\beta}_{1-42}\;A{\beta}_{1-43}$) to both SK-N-MC and PC12 cells. In the present study, we found that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and inhibitory potency to $\beta$-secretase activity. Conclusion : These results suggest that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol may contain the protective component(s) against $A\beta$-induced cell death in PC12 cells as well as inhibitory component(s) to $\beta$-secretase activity.

• 생약학회지
• /
• 제47권3호
• /
• pp.251-257
• /
• 2016

In this study, the radical scavenging activity and protective effect of ethanol extract from leaf of Engelhardtia chrysolepis HANCE (ECE) against oxidative stress were investigated under in vitro and cellular system. ECE showed strong radical scavenging activities in 1,1-diphenyl-2-picrylhydrazyl, hydroxyl(${\cdot}OH$) and nitric oxide(NO) radical as a concentration-dependent manner. Particularly, strong scavenging activity against the ${\cdot}OH$ and NO radical were observed with the $IC_{50}$ value of $1.30{\mu}g/ml$ and $12.61{\mu}g/ml$, respectively. Furthermore, the cellular oxidative stress was induced by amyloid beta($A{\beta}_{25-35}$) in C6 glial cells. The treatment of $A{\beta}_{25-35}$ to C6 glial cells generated high levels of reactive oxygen species(ROS) and declined cell viability. However, production of ROS was decreased by the treatment of ECE. In addition, the cell viability was significantly increased at each concentration(10, 25, $50{\mu}g/ml$) as dose-dependent manner. The Alzheimer's disease-related protein expressions in $A{\beta}_{25-35}$-treated C6 glial cells were analyzed. The ECE treatment inhibited expression of amyloid precursor protein(APP), C-terminal fragment-${\beta}(CTF-{\beta})$, ${\beta}$-site APP cleaving enzyme(BACE), phosphorylated tau(p-tau) proteins in C6 glial cells induced by $A{\beta}_{25-35}$. The present study indicated that ECE has strong radical scavenging activity and neuroprotective effect through attenuating oxidative stress.

• 동의신경정신과학회지
• /
• 제19권1호
• /
• pp.125-146
• /
• 2008

Objectives : The purpose of this study is to suggest for the following experimental study of dementia by reviewing recent oriental medicine journals that have been published since 2000. Methods: We have investigated various types of studies in relation to dementia through 90 articles that have been published from 2000 to 2007 in recent oriental medicine journals were registered Korea research foundation. Results and Conclusions : 1. Since 2000, 88 articles in relation to dementia have been published and almost of them are herbal medicine-centered studies. Also they show a tendency to increase every year. The journal of oriental neuropsychiatry carries the highest number of studies in relation to dementia. 2. According to the experimental paper, there are 30 cases of using herb simplexes, 48 cases of herb-combined prescription, and 10 cases of other ways. Especially 7 cases of using herb-combined prescription relation to Sasang constitution are all for the Taeumin. 3. There are 85 cases of Animal and cellular experimental, 60 cases of using pathologic model induced cytotoxic activity, a case of using L-NAME, 3 cases of 192 saporin, 4 cases of ibotenic acid, 10 cases of focal cerebral ischemia, 3 cases of alcohol-administered, and one case of natural degradation. 4. Moms water maze, Radial arm maze Passive avoidance learning model were using for examining learning and memory of model animal 5. We propose that following studies of dementia are to he investigated of the applied method of using siRNA with tranceduced gene, sample preparation by water-soaking, oriental medical diagnosis, standardization of differentiating symptom and herb simplexes, building the database by classified prescriptions, and experiment model which are based on precise examining mechanism with cell line as like mouse H19-7 hippocampus, rat HT22 hippocampus, astrocyte, microglia, using the model of animals at APP, PS1, BACE, CT99/PS1, APOE4, Tau, APP/PSI/Tau