Li, Naijing
(Department of Gerontology, The Shengjing Affiliated Hospital, China Medical University)
,
Liu, Ying
(College of Pharmacy, Shenyang Pharmaceutical University)
,
Li, Wei
(College of Pharmacy, Shenyang Pharmaceutical University)
,
Zhou, Ling
(College of Pharmacy, Shenyang Pharmaceutical University)
,
Li, Qing
(College of Pharmacy, Shenyang Pharmaceutical University)
,
Wang, Xueqing
(Department of Gastroenterology, The Shengjing Affiliated Hospital, China Medical University)
,
He, Ping
(Department of Gerontology, The Shengjing Affiliated Hospital, China Medical University)
Background: Alzheimer's disease (AD) is a progressive brain disease, for which there is no effective drug therapy at present. Ginsenoside Rg1 (G-Rg1) and G-Rg2 have been reported to alleviate memory deterioration. However, the mechanism of their anti-AD effect has not yet been clearly elucidated. Me...
Background: Alzheimer's disease (AD) is a progressive brain disease, for which there is no effective drug therapy at present. Ginsenoside Rg1 (G-Rg1) and G-Rg2 have been reported to alleviate memory deterioration. However, the mechanism of their anti-AD effect has not yet been clearly elucidated. Methods: Ultra performance liquid chromatography tandem MS (UPLC/MS)-based metabolomics was used to identify metabolites that are differentially expressed in the brains of AD mice with or without ginsenoside treatment. The cognitive function of mice and pathological changes in the brain were also assessed using the Morris water maze (MWM) and immunohistochemistry, respectively. Results: The impaired cognitive function and increased hippocampal $A{\beta}$ deposition in AD mice were ameliorated by G-Rg1 and G-Rg2. In addition, a total of 11 potential biomarkers that are associated with the metabolism of lysophosphatidylcholines (LPCs), hypoxanthine, and sphingolipids were identified in the brains of AD mice and their levels were partly restored after treatment with G-Rg1 and G-Rg2. G-Rg1 and G-Rg2 treatment influenced the levels of hypoxanthine, dihydrosphingosine, hexadecasphinganine, LPC C 16:0, and LPC C 18:0 in AD mice. Additionally, G-Rg1 treatment also influenced the levels of phytosphingosine, LPC C 13:0, LPC C 15:0, LPC C 18:1, and LPC C 18:3 in AD mice. Conclusion: These results indicate that the improvements in cognitive function and morphological changes produced by G-Rg1 and G-Rg2 treatment are caused by regulation of related brain metabolic pathways. This will extend our understanding of the mechanisms involved in the effects of G-Rg1 and G-Rg2 on AD.
Background: Alzheimer's disease (AD) is a progressive brain disease, for which there is no effective drug therapy at present. Ginsenoside Rg1 (G-Rg1) and G-Rg2 have been reported to alleviate memory deterioration. However, the mechanism of their anti-AD effect has not yet been clearly elucidated. Methods: Ultra performance liquid chromatography tandem MS (UPLC/MS)-based metabolomics was used to identify metabolites that are differentially expressed in the brains of AD mice with or without ginsenoside treatment. The cognitive function of mice and pathological changes in the brain were also assessed using the Morris water maze (MWM) and immunohistochemistry, respectively. Results: The impaired cognitive function and increased hippocampal $A{\beta}$ deposition in AD mice were ameliorated by G-Rg1 and G-Rg2. In addition, a total of 11 potential biomarkers that are associated with the metabolism of lysophosphatidylcholines (LPCs), hypoxanthine, and sphingolipids were identified in the brains of AD mice and their levels were partly restored after treatment with G-Rg1 and G-Rg2. G-Rg1 and G-Rg2 treatment influenced the levels of hypoxanthine, dihydrosphingosine, hexadecasphinganine, LPC C 16:0, and LPC C 18:0 in AD mice. Additionally, G-Rg1 treatment also influenced the levels of phytosphingosine, LPC C 13:0, LPC C 15:0, LPC C 18:1, and LPC C 18:3 in AD mice. Conclusion: These results indicate that the improvements in cognitive function and morphological changes produced by G-Rg1 and G-Rg2 treatment are caused by regulation of related brain metabolic pathways. This will extend our understanding of the mechanisms involved in the effects of G-Rg1 and G-Rg2 on AD.
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문제 정의
The aim of this study was to investigate the metabolic changes in the brains of G-Rg1 and G-Rg2 treated APP/PS1 transgenic mice. In addition, the Morris water maze (MWM) test and immunohistochemistry were used to assess the effects of G-Rg1 and G-Rg2 on the cognitive function and pathological changes in APP/PS1 mice.
In addition, the Morris water maze (MWM) test and immunohistochemistry were used to assess the effects of G-Rg1 and G-Rg2 on the cognitive function and pathological changes in APP/PS1 mice. To our knowledge, this is the first report of the use of a UPLC/MS-based brain tissue metabolomics method to investigate the interventional effects of G-Rg1 and G-Rg2 on AD, which may facilitate our understanding of the anti-AD mechanism of these ginsenosides.
제안 방법
Food and water were available ad libitum. After acclimatization for 1 wk, the mice were divided into four groups (n = 10 in each group): the normal control group, the AD model group, the G-Rg1 group, and the G-Rg2 group. According to the concentrationresponse curves, the mice in the G-Rg1 and G-Rg2 groups were injected intraperitoneally once daily with G-Rg1 and G-Rg2 (30 mg/ kg), respectively, dissolved in saline.
Mass spectrometric detection was performed on a Micromass Quattro Micro API mass spectrometer (Waters Corp.) with an electrospray ionization interface and triple quadrupole mass analyzer. The electrospray ionization source was set in the positive mode.
The animals, aged 7 mo, were maintained in an air-conditioned animal center at 23 ± 2 C and a relative humidity of 50 ± 10%, with a natural lightdark cycle.
3. The applied method was validated before the analysis of the experimental samples by determining the precision, withinday stability, and repeatability of sample preparation. The extracted ion chromatographic peaks of seven ions (m/z 132.
The applied method was validated before the analysis of the experimental samples by determining the precision, withinday stability, and repeatability of sample preparation. The extracted ion chromatographic peaks of seven ions (m/z 132.2, 0.52 min; m/z 120.1, 1.90 min; 710.5, 3.37 min; 346.7, 8.41 min; 502.3, 8.93 min; 374.1, 10.6 min; and 402.3, 12.88 min, in positive ion mode) were distributed across different spectral regions, and the retention times were selected to validate the method. The relative SDs (RSDs) of the peak intensities and retention times for the selected ions in the pooled brain sample were calculated.
7% for the peak intensity. The post-preparation stability of the sample was tested by analyzing a sample left in the autosampler (maintained at 4 C) for 4 h, 8 h, 12 h, and 24 h. The RSDs ranged from 3.
대상 데이터
Acetonitrile (HPLC grade) and methanol (HPLC grade) were purchased from Fisher Scientific (Fair Lawn, NJ, USA). Formic acid (HPLC grade) was obtained from Kermel Industrial (Tianjin, China).
데이터처리
The retention time and m/z were calculated for each peak. Principal components analysis (PCA) was used to select distinct variables, and Student t test was used to evaluate statistically significant differences between potential biomarkers (SPSS version19.0 software; SPSS Inc.). Statistical significance was accepted if p < 0.
SPSS version 19.0 software (SPSS Inc., Chicago, IL, USA) was used to analyze the data from the MWM test, which were evaluated by the analysis of variance, followed by Tukey’s multiple comparison test.
이론/모형
To determine whether the metabolites in brain differed between normal control and AD model mice, PCA, which is a classic unsupervised method (no prior knowledge concerning groups or tendencies within the data sets is necessary) of pattern recognition, was used. In the PCA score, each point represented an individual sample; the plot of PCA scores divided different brain samples into blocks, suggesting changes in the metabolic profiles.
성능/효과
The PCA model was also used to determine whether G-Rg1 and G-Rg2 affect the metabolite profiles of AD mice. According to the PCA score plots (Fig. 6A) from processing dates of the normal control, AD model, and G-Rg1 and G-Rg2 treated groups, samples from the ginsenosides treated groups were separated from the AD model group and were closer to the controls, suggesting that G-Rg1 and G-Rg2 treatment partly restored brain metabolites in AD mice to near normal levels. Metabolites with altered levels were identified in the PCA loading plot (Fig.
The MWM results show that the APP/PS1 mice were markedly deficient in cognitive function compared with age-matched C57BL/6J mice, which is consistent with previous studies [23]. Treatment with G-Rg1 and G-Rg2 effectively improved cognitive function of the mice that had declined due to AD.
In our previous study, we found changes in the sphingolipid levels in the plasma of AD patients [19]. The results of this study proved that disorders of sphingolipid levels are also observed in the brains of AD mice, and indicated that G-Rg1 and G-Rg2 also take part in sphingolipid metabolic pathways. G-Rg1 treatment increased the levels of dihydrosphingosine, hexadecasphinganine, and phytosphingosine, whereas G-Rg2 treatment just increased the levels of dihydrosphingosine and hexadecasphinganine.
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