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1.
Article in Chinese | WPRIM | ID: wpr-940213

ABSTRACT

ObjectiveTo study the inhibitory effect of aloe-emodin (AE) on aluminum ion (Al3+)-induced β-amyloid protein 42 (Aβ42) aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro, and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group, Aβ42+Al3+ group, Aβ42+AE group, Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process, aggregation morphology, aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T (ThT) fluorescence assay, transmission electron microscopy (TEM), dynamic light scattering (DLS) experiment and thiazolyl blue (MTT) cytotoxicity assay. ResultCompared with the Aβ42 group, Al3+ could promote Aβ42 aggregation, increase the fluorescence intensity of ThT by 124.48%, induce the aggregation of Aβ42 to form fiber bundles with larger particle size, and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells (P<0.01), thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation, but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group, high concentration of AE could reduce the ThT fluorescence intensity to 41.66%, and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides, it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ (P<0.01), and restored the cell survival rate to 84.87%. Further depolymerization was conducted, AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+, depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity, thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.

2.
Article in Chinese | WPRIM | ID: wpr-940116

ABSTRACT

ObjectiveTo study the inhibitory effect of aloe-emodin (AE) on aluminum ion (Al3+)-induced β-amyloid protein 42 (Aβ42) aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro, and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group, Aβ42+Al3+ group, Aβ42+AE group, Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process, aggregation morphology, aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T (ThT) fluorescence assay, transmission electron microscopy (TEM), dynamic light scattering (DLS) experiment and thiazolyl blue (MTT) cytotoxicity assay. ResultCompared with the Aβ42 group, Al3+ could promote Aβ42 aggregation, increase the fluorescence intensity of ThT by 124.48%, induce the aggregation of Aβ42 to form fiber bundles with larger particle size, and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells (P<0.01), thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation, but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group, high concentration of AE could reduce the ThT fluorescence intensity to 41.66%, and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides, it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ (P<0.01), and restored the cell survival rate to 84.87%. Further depolymerization was conducted, AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+, depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity, thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.

3.
Article in Chinese | WPRIM | ID: wpr-927891

ABSTRACT

Objective: To uncover the time-dependent expression pattern of ptk2b gene and ptk2b-encoded protein, protein tyrosine kinase 2 beta(PTK2B), in the brain tissues of transgenic animal models of Alzheimer's disease (AD) and its relationship with the levels of Aβ1-42, phosphorylation of Tau (p-Tau) and low density lipoprotein receptor-related protein-1(LRP-1) in blood and brain tissues. Methods: In this study, 5-, 10- and 15-month-old APPswe/PS1dE9 double-transgenic mice harboring the genotype of AD confirmed by the gene test were divided into the 5-, 10- and 15-month-old experiment groups, and simultaneously, age-matched C57BL/6J mice were placed into the corresponding control groups, with 8 mice in each group. All mice were subjected to the Morris Water Maze for test of cognitive and behavioral ability. Expression profiles of PTK2B, Aβ1-42, p-Tau/Tau and LRP-1 in the hippocampus or blood of mice were quantified by using the immunohistochemistry staining, Western blot or enzyme-linked immunosorbent assay (ELISA), while the mRNA expression of ptk2b in the hippocampus was quantified by using the real-time quantitative polymerase chain reaction (qRT-PCR). Results: Results of experiment groups demonstrated that as mice aged, the expression levels of PTK2B, ptk2b mRNA, Aβ1-42 and p-Tau/Tau in the hippocampus were increased, and the expression of LRP-1 was decreased gradually. While in the blood, the level of Aβ1-42 was decreased, and the cognitive and behavioral ability was decreased in an age-dependent manner (all P< 0.05). However, comparisons among the control groups, only the age-dependent downregulation of LRP-1 were observed in hippocampus(P<0.05), but other indicators had no significant differences (P>0.05). Conclusion: In the hippocampus of APP/PS1 double-transgenic mice, the expressions of PTK2B, Aβ1-42 and p-Tau/Tau are upregulated, LRP-1 is downregulated, while cognitive and behavioral ability is decreased, and such changes are presented in a time-dependent manner.


Subject(s)
Alzheimer Disease/metabolism , Amyloid beta-Peptides , Amyloid beta-Protein Precursor/genetics , Animals , Focal Adhesion Kinase 2/metabolism , Hippocampus/metabolism , Low Density Lipoprotein Receptor-Related Protein-1 , Maze Learning , Mice , Mice, Inbred C57BL , Mice, Transgenic , RNA, Messenger
4.
Chinese Acupuncture & Moxibustion ; (12): 1231-1235, 2021.
Article in Chinese | WPRIM | ID: wpr-921037

ABSTRACT

OBJECTIVE@#To compare the clinical efficacy of abdominal acupoint thread embedding therapy based on "brain-intestinal connection" combined with donepezil hydrochloride tablets and oral donepezil hydrochloride tablets alone for mild-to-moderate Alzheimer's disease (AD) and observe its effects on amyloid precursor protein (APP) and β-amyloid protein@*METHODS@#Sixty patients with AD were randomly divided into an observation group (30 cases, 3 cases dropped off) and a control group (30 cases, 3 cases dropped off). The patients in the control group were treated with donepezil hydrochloride tablets (5 mg per day); based on the treatment in the control group, the patients in the observation group were treated with abdominal acupoint thread embedding therapy at Zhongwan (CV 12), Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Daheng (SP 15), etc., once every 10 days. Both groups were treated for 2 months. The mini-mental state examination (MMSE), Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), activity of daily living scale (ADL), neuropsychiatric inventory questionnaire (NPI) as well as the serum levels of APP and Aβ@*RESULTS@#After treatment, the MMSE scores in the two groups were higher than those before treatment (@*CONCLUSION@#The abdominal acupoint thread embedding therapy based on the theory of "brain-intestinal connection" combined with donepezil hydrochloride tablets can improve cognitive function, self-care ability of daily life and mental behavior, and reduce the serum levels of APP and Aβ


Subject(s)
Acupuncture Points , Alzheimer Disease/drug therapy , Amyloid beta-Peptides , Amyloid beta-Protein Precursor , Brain , Donepezil , Humans , Peptide Fragments
5.
Acupuncture Research ; (6): 426-431, 2020.
Article in Chinese | WPRIM | ID: wpr-844151

ABSTRACT

Alzheimer's disease (AD), a neurodegenerative disorder characterized by amyloid deposits and neurofibrillary degeneration, is the most common type of dementia and has no incurable therapies at the moment. Electroacupuncture (EA) therapy has been widely used in clinical treatment of AD, and has attained approving effects. This article reviews the development of researches on the mechanisms of EA underlying improving AD by diminishing β amyloid protein (Aβ) neurotoxicity, from 1) up-regulating hippocampal cellular autophagy, 2) improving cerebral energy metabolism by activating oxidation stress-related factors peroxisome proliferator-activated receptor γ coactivator 1 alpha and sirtuin 1 in the hippocampus and frontal cerebral cortex, 3) relieving inflammatory reaction by lowering expression of tumor necrosis factor-alpha and high-mobility group box 1 and increasing expression of Interleukin 10, and 4) promoting degradation of Aβ1-42 by down-regulating expression of insulin degeneration enzyme, lipoprotein, transthyretin, apolipoprotein and α2 mcroglobulin. Meanwhile, a comprehensive clinical therapy of AD is proposed.

6.
Article in Chinese | WPRIM | ID: wpr-872764

ABSTRACT

Objective:To investigate the effects of naringenin on oxidative stress and Tau protein phosphorylation of adrenal pheochromocytoma(PC12) cells injured by β-amyloid(Aβ)25-35 and its relationship with estrogen receptor(ER) and phosphatidylinositol -3 kinase/protein kinase B(PI3K/Akt) signaling pathway. Method:The PC12 cells were intervened with Aβ25-35 to prepare the injury model. The experiment was divided into blank group, model group, naringenin(400,40,4,0.4,0.04,4×10-3,4×10-4,4×10-5 μmol·L-1)group, positive drugs estradiol(E2)(1 nmol·L-1)+Aβ25-35 group, naringenin(0.4,0.04,4×10-3,4×10-4,4×10-5 μmol·L-1)+Aβ25-35 group, E2+Aβ25-35+ER antagonist(ICI182780)(1 μmol·L-1) group, naringenin+Aβ25-35+ICI182780 group, E2+Aβ25-35+PI3K blocker(LY294002)(50 μmol·L-1) group, naringenin+Aβ25-35+LY294002 group. Methye thiazolye telrazlium(MTT)method was used to detect the cell proliferation index, 2',7'-Dichlorodi -hydrofluorescein diacetate(DCFH-DA) was used as a fluorescent probe to detect the content of reactive osygen species(ROS), the content of malondialdehyde(MDA) and the activity of superoxide dismutase(SOD) were measured by thiobarbituric acid(TBA) and oxidase methods, Western blot was used to detect the expression of phosphorylated Tau protein/total Tau protein(p-Tau/t-Tau). Result:According to the results of MTT experiment, 0.4 μmol·L-1 was selected as the best effective concentration of naringenin, compared with the blank group, the cell proliferation index of model group decreased significantly (P<0.01), compared with model group, the cell proliferation index of naringenin+Aβ25-35 group increased significantly (P<0.01). In addition, compared with blank group, the content of ROS, MDA and the expression of p-Tau/t-Tau in the model group increased significantly (P<0.01), and the activity of SOD decreased significantly (P<0.01), compared with model group, the content of ROS, MDA and the expression of p-Tau/t-Tau in naringenin+Aβ25-35 group decreased significantly (P<0.01), and the activity of SOD increased significantly (P<0.01), compared with naringenin+Aβ25-35 group, the addition of ICI182780 and LY294002 significantly reversed the role of naringenin in the above indicators (P<0.01). The effect of naringenin was similar to that of E2. Conclusion:Naringenin can improve the cell proliferation index and protect PC12 cells from Aβ25-35 injury, which may be achieved by activating ER and PI3K/Akt signaling pathway to reduce ROS, MDA content, p-Tau/t-Tau expression and promote SOD activity.

7.
China Pharmacy ; (12): 2519-2523, 2020.
Article in Chinese | WPRIM | ID: wpr-829361

ABSTRACT

OBJECTIVE:To investigate the mechanism of sinapic acid (SA)against PC 12 cell injury induced by Amyloid β1-42 protein(Aβ1-42)based on PI 3K/Akt/GSK3β signaling pathway. METHODS:PC12 cells of rats were randomly divided into control group,Aβ group(Aβ1-42 2 μmol/L),Aβ+SA group(Aβ1-42 2 μmol/L+SA100 μmol/L),Aβ+SA+LY group [Aβ1-42 2 μmol/L+SA 100 μmol/L+LY294002(PI3K inhibitor )10 μmol/L],Aβ+LY group(Aβ1-42 2 μmol/L+LY294002 10 μmol/L)and LY group (LY294002 10 μmol/L). Except for control group and LY group ,the cells of other groups were replicated the damage model with Aβ1-42. After 24 hours of culture ,the morphology of cells was obsened in each group with a microscope ,and MTT assay was adopted to determine the cell viability of PC 12 cells in each group. Western blotting assay was used to detect the expression of PI 3K,p-PI3K, Akt,p-Akt,GSK3β and p-GSK3β in cells of each group. RESULTS:Compared with control group ,the number of cells decreased and some synaptic breaks disappeared in Aβ group while cell viability,ratio of p-PI 3K/PI3K,p-Akt/Akt and p-GSK 3β/GSK3β in Aβ group were decreased significantly(P<0.05 or P<0.01). Compared with Aβ group,the cells became round and synapses became more in Aβ+SA group while cell viability,the ratio of p-PI 3K/PI3K,p-Akt/Akt and p-GSK 3β/GSK3β were increased significantly(P<0.05). Compared with Aβ+SA group,some synaptic breaks occurred in Aβ+SA+LY group while cell viability, the ratio of p-PI 3K/PI3K,p-Akt/Akt and p-GSK 3β/GSK3β were decreased significantly(P<0.05);Aβ+LY group had more cell debris,and t he cell viability was decreased ,but the difference was not significant ,and the ratio of p-PI 3K/PI3K,p-Akt/Akt and p-GSK3 β/GSK3 β had no significant change (P>0.05); LY294002 alone had no significant effect on morphology ,cellviability and the ratio of p-PI 3K/PI3K,p-Akt/Akt or p-GSK 3β/ GSK3 β (P>0.05). CONCLUSIONS : SA may play aprotective role against PC 12 cell injury induced by A β 1-42 through activating PI 3K/Akt/GSK-3β.

8.
Chinese Pharmacological Bulletin ; (12): 539-543, 2020.
Article in Chinese | WPRIM | ID: wpr-856999

ABSTRACT

Aim To explore mechanism of epigallocatechin-3-gallate (EGCG) on alleviation of hippocampal neuronal autophagy in APP/PSI transgenic mice. Methods 8-month old APP/PSI transgenic mice were randomly divided into three groups;model group (Tg), EGCG low dose group (Tg/EGCG-L), high dose group (Tg/EGCG-H). C57BL/6J mice were utilized as control. Learning and memory were detected by Morris water maze test. The hippocampal ULK1, P62, LC3 I I / LC3 I,mT0R and Aß M2 expressions were detected by Western blot, immunohistochemical staining and ELISA. Results Compared with NT mice, Tg mice showed a marked prolongation of the escape latency in MWM test (P <0. 05). Decreased ULK1 expression and increased P62, LC3 II/LC3 I and A ßM 2 were detected (P < 0. 05). EGCG-treated group showed marked improvement of all these abnormal changes (P < 0. 05). Conclusions EGCG treatment is able to improve cognitive function, which may be attributed to ameliorated autophagic networks dysfunction and reduced Aß plaques in the the hippocampi of APP/PS1 transgenic mice.

9.
Electron. j. biotechnol ; 40: 1-9, July. 2019. tab, graf, ilus
Article in English | LILACS | ID: biblio-1053195

ABSTRACT

BACKGROUND: Microalgae are aquatic chlorophyll-containing organisms comprising unicellular microscopic forms, and their biomasses are potential sources of bioactive compounds, biofuels and food-based products. However, the neuroprotective effects of microalgal biomass have not been fully explored. In this study, biomass from two Chlorella species was characterized, and their antioxidant, anticholinesterase and anti-amyloidogenic activities were investigated. RESULTS: GC­MS analysis of the extracts revealed the presence of some phenols, sterols, steroids, fatty acids and terpenes. Ethanol extract of Chlorella sorokiniana (14.21 mg GAE/g) and dichloromethane extract of Chlorella minutissima (20.65 mg QE/g) had the highest total phenol and flavonoid contents, respectively. All the extracts scavenged 2,2-diphenyl-1-picrylhydrazyl, 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate) and hydroxyl radicals. The highest metal chelating activity of the extracts was observed in the ethanol extracts of C. minutissima (102.60 µg/mL) and C. sorokiniana (107.84 µg/mL). Furthermore, the cholinesterase inhibitory activities of the extracts showed that ethanol extract of C. sorokiniana (13.34 µg/mL) exhibited the highest acetylcholinesterase inhibitory activity, while dichloromethane extract of C. minutissima (11.78 µg/mL) showed the highest butyrylcholinesterase inhibitory activity. Incubation of the ß-amyloid protein increased the aggregation of amyloid fibrils after 96 h. However, ethanol extract of C. sorokiniana and C. minutissima inhibited further aggregation of Aß1­42 and caused disaggregation of matured protein fibrils compared to the control. This study reveals the modulatory effects of C. sorokiniana and C. minutissima extracts on some mediators of Alzheimer's disease and provides insights into their potential benefits as functional food, nutraceutics or therapeutic agent for the management of this neurodegenerative disease.


Subject(s)
Chlorella/chemistry , Cholinesterase Inhibitors/pharmacology , Amyloid beta-Peptides/antagonists & inhibitors , Antioxidants/pharmacology , Phenols/analysis , Steroids/analysis , Sterols/analysis , Terpenes/analysis , Plant Extracts/pharmacology , Plant Extracts/chemistry , Cholinesterase Inhibitors/chemistry , Spectroscopy, Fourier Transform Infrared , Neuroprotective Agents , Biomass , Ethanol , Fatty Acids/analysis , Microalgae , Alzheimer Disease/prevention & control , Amyloid/drug effects , Gas Chromatography-Mass Spectrometry , Antioxidants/chemistry
10.
Article in Chinese | WPRIM | ID: wpr-801964

ABSTRACT

Objective: To investigate the neuroprotective effect and mechanism of tetrahydroxy stilbene glucoside (TSG) on β-amyloid protein 25-35 (Aβ25-35)-induced neuron synapses damage. Method: Primary neurons were isolated and purified from cerebral cortex of suckling mouse. Then neurons were divided into control group, model group (incubation with Aβ25-35) and TSG groups (after incubation with Aβ25-35, add 6.25, 12.5, 25, 50, 100 μmol·L-1 TSG). Cell counting kit-8 (CCK-8) and Lactate dehydrogenase (LDH) methods were used to observe the viability of neuron, immunocytochemical staining was performed to determine the expressions of synapsin-1 (SYN-1), and the concentration of postsynaptic density-95 (PSD-95) and synaptophysin (SYP) were detected by enzyme-linked immunosorbent assay (ELISA) method. The level of cyclic adenosine monophosphate response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) and the level of CREB, Phosphorylated CREB (p-CREB) and BDNF proteins were determined by immunocytochemical staining or Western blot (WB). Result: Compared with normal group, the cell survival rate of model group was significantly reduced, LDH release was significantly increased (PPPPPPP-1,25 μmol·L-1 TSG can significantly enhance the expression of SYN-1(PPPPConclusion: TSG possesses the neuroprotective effect on Aβ25-35-induced neuron synapses, the mechanism may be associated with the activation of CREB/BDNF signaling pathway.

11.
Chinese Pharmaceutical Journal ; (24): 703-710, 2019.
Article in Chinese | WPRIM | ID: wpr-858016

ABSTRACT

OBJECTIVE: To discuss the effect of ferulic acid(FA) on learning and memory impairment and neuron protection by repairing the imbalance of mitochondrial fission-fusion dynamics in Alzheimer′s disease (AD) mice. METHODS: The KM mice were randomly divided into normal control group (A group, n=10), model group (B group, n=10), positive control group (huperzine A tablets, C group, n=10) and low dose of FA group (D-low group, n=10), high dose of FA group (D-high group, n=10). Mice in B, C, D-low and D-high groups were built as AD model by injecting Aβ1-42 into the lateral ventricle. The learning and memory ability of mice were detected by Morris water maze test. The mRNA of dynamin-related protein 1 (Drp1) were detected by PCR. The AD related pathological proteins and mitochondrial fission-fusion proteins were detected by Western blot. The content and distribution of Aβ was analyzed using immunofluorescence staining. RESULTS: ①The escape latency of mice in D-high group was shorter than B group, but a little longer than A group (P0.05). ②The mean expressions of Drp1, CaN subunit α (CnAα), CnAβ mRNA in D-high group were lower than B group, but higher than A group (P<0.05). ③The mean expressions of amyloid precursor protein (APP), beta-site APP cleaving enzyme 1 (Bace1), Tau46 and pS396 proteins in D-high group were lower than B group, but higher than A group (P<0.05).The mean expressions of Drp1Ser637, CnAα, protein kinase A catalytic subunit c (PKAc), mitofusin gene 2(Mfn2) proteins in D-high group were basically identical with A group. ⑤The levels of Aβ formation and accumulation in mice cortex and hippocampus of D-high group were less than B group. CONCLUSION: It′s suggested that ferulic acid(FA) might repair pathological damage of Alzheimer′s disease by improving the imbalance of mitochondrial fission-fusion dynamics.

12.
Article in Chinese | WPRIM | ID: wpr-753931

ABSTRACT

Objective To investigate the relationship between serum markers β amyloid (Aβ), tau and thyroid hormone levels and post-stroke cognitive impairment (PSCI) in the acute phase of cerebral infarction. Methods A total of 214 patients with acute cerebral infarction were enrolled. The baseline data and serological indicators were collected and the cognitive function of patients was evaluated. All patients were divided into cognitive impairment group and normal group based on follow-up results. The differences of Aβ1-42, tau protein and thyroxine levels between the two groups and their relationship with disease progression were analyzed. The Cox regression analysis and ROC curve were used to compare the above parameters to predict the development of PSCI. Results The total protein level of Tau (210.6 ±98.9 pg/mL) was higher and Aβ1-42 (426.1 ±123.5 pg/mL) and triiodothyronine (T3) (1.43 ±0.57 nmol/L), free thyroxine (FT4) (13.15±2.23 pmol/L) was significantly lower in the cognitive impairment group than in the normal group (P<0.05). Tau protein (r=-0.457), Aβ1-42 (r=0.348), T3 (r=0.211), and FT4 (r=0.306) were all associated with disease progression (P<0.05). Cox regression analysis showed that Aβ1-42 and T3 were important influencing factors in the occurrence of PSCI. The area under the curve of Aβ1-42 combined with T3 was 0.841. The specificity and the sensitivity were 74.8% and 85.3%, respectively, with a diagnostic cutoff value of 0.572. Conclusion Aβ1-42 and T3 levels in the acute phase of cerebral infarction may predict the progression of PSCI.

13.
Article in Chinese | WPRIM | ID: wpr-861349

ABSTRACT

Alzheimer disease (AD) is a common neurodegenerative disorder in the elderly. It is the most common cause of dementia and is difficult to diagnose in the early stage. PET imaging has important value for early diagnosis, and amyloid imaging is the best choice for early diagnosis of AD, glucose imaging has great value of assessment of disease condition. The application and progresses of PET imaging in early diagnosis of AD, focusing on glucose imaging, amyloid protein imaging and tau protein imaging were reviewed in this article.

14.
Chinese Journal of Geriatrics ; (12): 324-329, 2018.
Article in Chinese | WPRIM | ID: wpr-709249

ABSTRACT

Objective To investigate the effect of ten-eleven translocation protein on the proliferation of human neuroblastoma cell lines SH-SY5Y and IMR 32 and the expression of amyloid precursor protein,PS1,and β site APP cleaving enzyme 1 in the absence of folic acid and possible mechanisms involved.Methods SH-SY5Y and IMR-32 cells were cultured in vitro and divided into the folic acid deficiency group (0 mg/L),the low folic acid group (1mg/L),and the normal control group (4mg/L).The MTT method was used to observe cell proliferation,and RT-PCR was adopted to detect the mRNA expression of APP,PS1,BACE1,DNMTs and TETs in the cells in real-time.Besides,we generated a stable low-level TET1 expression cell line,and compared the expression with that in a negative control group.Furthermore,the expression of fluorescent protein was observed by fluorescence inverted microscope,cell proliferation was measured by the MTT assay,and mRNA levels of TET1,APP,PS1,and BACE1 were detected by RT-PCR.Results (1) In the folic acid deficiency group and the low folic acid group,cell proliferation of SH-SY5Y after 120 h and of IMR-32 cell after 144 h significantly decreased (P<0.001).The mRNA levels of APP,PS1,BACE1,DNMT1,DNMT3a,DNMT3b,TET1,TET2,and TET3 in SH-SY5Y cells increased (F=80.315,35.386,101.979,786.407,80.331,131.545,28.000,9.165,and 102.167,all P<0.05);the mRNA levels of APP,PS1,BACE1,DNMT1,DNMT3b,TET1,TET2,and TET3 in IMR-32 cells also rose (F=12.283,93.669,40.815,157.234,24.835,147.594,54.794,and 73.068,all P<0.05).(2) Generation of a stable low-level TET1 expression cell line:The mRNA level of TET1 in the low expression group (SH-SY5Y-shTET1) was 0.25± 0.02,which was significantly lower than that in the negative control group (1.00±0.09) (P=0.007);the mRNA level of TET1 in the low expression group (IMR-32-shTET1) was 0.28 ±0.07,significantly lower than that in the negative control group (1.00±0.01) (P=0.003).(3)The proliferative ability of the low expression groups (SH-SY5Y-shTET1 and IMR-32-shTET1) was significantly higher than that in the negative control group (P<0.01).The mRNA levels of APP and BACE1 decreased (P<0.01 or P<0.05)Conclusion In the human neuroblastoma cell lines SH-SY5Y and IMR-32,folic acid deficiency up-regulates the expression of TETs,increases the expression of APP and BACE1 in the cells by TET protein demethylation,and inhibits cell growth.

15.
Article in Chinese | WPRIM | ID: wpr-704993

ABSTRACT

Objective To investigate the expression and relationship of canonical transient receptor potential channel-3 (TRPC3) and brain-derived neurotrophic factor (BDNF) in the hippocampus of a rat model of Alzheimer's disease (AD). Methods SD rats were randomly divided into PBS, AD, and AD+BDNF experimental groups. AD models were generated by intracerebroventricular injection ofβ-amyloid protein (Aβ1-42). BDNF was injected via the lateral ventricle catheter after 14 days. The Morris water maze test was used to assess the spatial learning and memory ability of the rats. The expression of TRPC3 and BDNF mRNA and protein in the hippocampus were detected by RT-PCR and Western blotting, respectively. Results The Morris water maze test showed that the escape latencies of the fifth day in the AD group were longer than those in the PBS group (P < 0. 05). The escape latencies in the AD+BDNF group were shorter than those in the AD group (P < 0. 05). RT-PCR and Western blotting results showed that the expression of both TRPC3 and BDNF were reduced in the AD group compared with the PBS group (P < 0. 05). TRPC3 expression was increased in the AD+BDNF group compared with the AD group (P < 0. 05). Conclusion The expression of BDNF and TRPC3 is decreased in the hippocampus of AD rats. An exogenous BDNF injection appears to improve the spatial learning and memory of AD rats that are impaired by a Aβ1-42 injection, possibly via TRPC3 upregulation, and may play a protective role in neurons.

16.
Article in Chinese | WPRIM | ID: wpr-703235

ABSTRACT

Objective To observe the pathological changes of brain tissues in the WHBE rabbit model of sporadic Alzheimer disease (AD). Methods Thirty 3 -4-month old male WHBE rabbits were randomly divided into 3 groups:normal control (NC) group, high cholesterol diet (HCD) group, high cholesterol diet + copper drinking water ( HCD+ Cu2+) group, 10 in each group. Another 10 senile (36-48-month old) male WHBE rabbits were taken as senile group. The NC group and the senile group were fed a normal basic diet, the HCD group fed a 2% cholesterol diet, and the HCD+Cu2+group fed a 2% cholesterol diet plus 0. 12 PPM copper drinking water for 12 weeks. The levels of total cholesterol (TC) and β-amyloid protein (Aβ) 1-42 were measured at 12 weeks. The activity of superoxide dismutase ( SOD) and the content of malondialdehyde (MDA) in the cortex and the hippocampus were detected. In addition, the covered area of Aβ, β-site APP cleaving enzyme 1(BACE1) and phosphorylated tau (p-tau) protein in coronal sections of brain tissues were also observed by immunohistochemical staining. The senile plaques and the neurofibrillary tangles were observed by Congo red and Bielschowsky staining, respectively. Results The body weight of WHBE rabbits in the senile group was significantly higher than that of the NC group ( P < 0. 01 ), and the plasma TC and Aβ1 -42 in each group were significantly higher than that in the NC group (P < 0.05, P < 0.01). The activity of SOD in brain tissues was significantly lower than that of NC group (P< 0. 05), and the MDA content was significantly higher than that of NC group (P< 0. 05, P < 0. 01). Immunohistochemical staining showed that the covered area of Aβ, BACE1 and p-tau in brain tissues of all groups were significantly higher than that of NC group (P < 0. 05, P < 0. 01), and the covered area of BACE1 and p-tau protein in the brain tissues of HCD + Cu2+group was also significantly higher than that of the HCD group (P < 0. 05, P < 0. 01). Congo red and Bielschowsky staining showed that the number of senile plaques and neurofibrillary tangles were observed in the brain tissues of the HCD, HCD+Cu2+and senile groups. Conclusions High cholesterol diet or supplemented with trace copper drinking water can induce obvious AD pathological changes in WHBE rabbit models of sporadic AD with obvious oxidative damage, increased Aβ deposition and senile plaque in the brain, and pathological changes of tau. WHBE rabbit can be used in the study of animal models of neurodegenerative diseases.

17.
Chinese Journal of Pathophysiology ; (12): 168-172,182, 2018.
Article in Chinese | WPRIM | ID: wpr-701096

ABSTRACT

AIM:To investigate the effects of curcumin on the viability ,the lactate dehydrogenase(LDH)re-lease,the apoptosis,and the activity and the expression levels of caspase-3,caspase-8 and caspase-9 of rat adrenal pheo-chromocytoma PC12 cells induced by β-amyloid protein 25-35(Aβ25-35 ).METHODS:The PC12 cells were treated with Aβ25-35.The viability and LDH release rates were measured by MTT assay and LDH kit ,respectively.The cells were ran-domly divided into blank control group ,model group,curcumin 10μmol/L group and curcumin 20μmol/L group.The ap-optotic rates were evaluated by flow cytometry with Annexin V-FITC/PI staining.The activities and expression levels of caspase-3,caspase-8 and caspase-9 were detected by colorimetric method and Western blot analysis.RESULTS:Com-pared with model group ,curcumin significantly increased the viability ,and decreased the LDH release rates and the apop-totic rates of the PC12 cells treated with Aβ25-35(P<0.01).Compared with model group,curcumin significantly decreased the activity and expression levels of caspase-3,caspase-8 and caspase-9(P<0.05 or P<0.01).CONCLUSION:Cur-cumin inhibits Aβ25-35-induced apoptosis of PC12 cells by inhibiting the expression of caspase-3,caspase-8 and caspase-9.

18.
Article in Chinese | WPRIM | ID: wpr-851515

ABSTRACT

To study the effects of prescriptions including Ginseng, DabuyuanDecoction, and XixinDecoctionfor tonifying spleen and stomach on the activity and migration ability of neural stem/progenitor cells (NSPCs) induced by β-amyloid protein (β-amyloid),and investigate the effect of hippocampal neuronal repair and regeneration. Methods NSPCs were isolated from fetal rats of 14-16 d gestation and identified. It was randomly divided into seven groups: blank group, blank group treated with Aβ1-42, control group of normal cerebrospinal fluid, control group of Aricept, prescription of Tonifying Spleen and Stomach and Yuanqi (Ginseng, Dabu Yuan Decoction, and XixinDecoction) Protection groups. The third generation of neural stem cells were treated with 25 μmol/L Aβ1-42 for 48h. The activity of NSPCs in each group was detected bytranswell assay and CCK-8 assay and themigration was detected by scratch test. The content of reactive oxygen species (Ros) in the cells was detected by flow cytometry. ResultsBy immunofluorescence staining, NSPCs specific protein Sox2 staining was positive. The CCK-8 detection results showed that the activity of NSPCs in hippocampus was significantly increased in the groups of Tonifying Spleen and Stomach YuanqiPrescription (P< 0.01); The results of Transwell experiment and scratch test showed that the migration ability of NSPCs was significantly increased (P< 0.05, 0.01). The flow cytometryshowed that the content of ROS in the NSPCs of spleen and stomach prescription groups decreased significantly (P< 0.01). Conclusion The prescriptionsfor invigorating spleen and stomach promoted the activity and migration of NSPCs in rat hippocampal neural stem cells. The oxidative stress induced by Aβcan be antagonized and its active ingredients can easily penetrate the blood-brain barrier to exerta stronger neuroprotective effect.

19.
Acupuncture Research ; (6): 692-697, 2018.
Article in Chinese | WPRIM | ID: wpr-844380

ABSTRACT

OBJECTIVE: To observe the effect of acupuncture and moxibustion (AM) on learning-memory ability and expression of amyloid beta (Aβ) in the hippocampal dentate gyrus (DG) of Alzheimer's disease (AD) rats, so as to explore its mechanism underlying improvement of AD. METHODS: Forty male Wistar rats were randomly divided into normal, sham operation, model and AM groups (n=10 in each). The AD model was established by bilateral hippocampal injection of Aβ1-42(5 µL). The AM was applied at "Baihui" (GV 20) and "Shenshu" (BL 23) for 15 min, once daily for 12 times. Morris water maze tests were used to assess the rats' learning-memory ability. The levels of serum Aβ1-42 and Aβ internalizing enzymes including transthyretin (TTR), lipoprotein lipase (LPL), alpha 2 macroglobulin (α 2M) and apolipoprotein E (ApoE) were detected by ELISA. The expression of Aβ1-42 in the hippocampal DG was detected by immunohistochemistry. RESULTS: Compared with the sham operation group, the average escape latency of location navigation test was significantly prolonged in the first 5 days and the last 3 days (P0.05). CONCLUSION: AM can improve the learning-memory ability of AD rats, which may be related to its effects in up-regulating the contents of serum Aβ internalizing enzymes and promoting the clearance of hippocampal Aβ. It suggests a protective role of AM on hippocampal neurons.

20.
International Eye Science ; (12): 35-39, 2018.
Article in Chinese | WPRIM | ID: wpr-695116

ABSTRACT

AIM:To observe the mechanism of Tianma Gouteng Decoction on the protein molecular level in the optic nerve crush model rats.METHODS:Totally 36 participants 36 male Wistar rats were divided randomly into six groups(6 in every group):normal control group,negative control group,Tianma Gouteng Decoction treatment groups (con-centrations were 0.6g/mL,1.2g/mL,2.4g/mL respictively) and ginkgo biloba tablets positive control group (concentrations was 1.2mg/mL).Nothing was done in the normal control group.The optic nerve of right eye in the other groups was done with the optic nerve crush model.Normal control group and negative control group was treated only with water.The average grey scale values of the N-methyl-D-aspartic acid receptor 2B (NMDA2B) receptor protein,beta-amyloid protein (Aβ) in the average grey scale values were detected.RESULTS:The average grey scale value of Tianma Gouteng Decoction in low,medium and high dose groups about NMDA2B receptor protein was significantly less than that of the negative control group (all P<0.001),and there was no significant difference with the positive control group (P=0.092,0.411,0.676),the difference between normal control group and negative control group was significant (P<0.001).The high dose group of betaamyloid's average grey scale value reduced significantly than the negative control group (P=0.030,0.001).The low dose group than the negative control group was not obviously (P=0.614).The high dose group was not significantly different from the positive control group (P=0.927),the difference between normal control group and negative control group was significant (P<0.001).CONCLUSION:Tianma Gouteng Decoction can go through the decrease of the NMDA2B receptor protein expression and the control of beta-amyloid deposition to reduce the retinal ganglion cell injury and apoptosis.

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