ABSTRACT
Objective To apply high performance liquid chromatography quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS/MS) method in analyzing and identifying the main components in the ethanol extract of Choeros pondiatis fructus. Methods The ethanol extract of Choeros pondiatis fructus was prepared by the refluxing methods, and the main components in the extracts were separated by the gradient elution method with Agilent TC C18 column (250 mmX4. 6 mm, 5 μm), methanol-0. 2% formic acid as the mobile phase, at flow rate 0. 7 mL/min, column temperature 30°C, and sample volume 25 μL Mass spectrometry was applied for the qualitative analysis under the positive and negative ion modes and ESI ion source was used for mass spectra, Vcap 4. 0 kV, drying gas 10 L/min, gas temperature 350°C. and nebulizer 35 psi (1 psi=6 894. 8 Pa). The main chromatographic peaks were assigned by Q-TOF. Results Forty-lwo compounds were identified through direct comparison of both positive and negative ion mass data, the clement composition analysis, the data of the literature and the information from reference substances, including amino acids, organic acids, and flavonoids. Among them 17 constituents have been firstly found in Choerospondiatis fructus. Conclusion HPLC-Q-TOF-MS/MS has the advantage of the efficient separation ability of HPLC and highly sensitive detection of MS, and is capable of making a qualitative analysis of the main components in Choerospondiatis fructus, which provides an evidence for study.
ABSTRACT
<p><b>BACKGROUND</b>Amyloid β (Aβ) deposits and the endoplasmic reticulum stress (ERS) are both well established in the development and progression of Alzheimer's disease (AD). However, the mechanism and role of Aβ-induced ERS in AD-associated pathological progression remain to be elucidated.</p><p><b>METHODS</b>The five familial AD (5×FAD) mice and wild-type (WT) mice aged 2, 7, and 12 months were used in the present study. Morris water maze test was used to evaluate their cognitive performance. Immunofluorescence and Western blot analyses were used to examine the dynamic changes of pro-apoptotic (CCAAT/enhancer-binding protein homologous protein [CHOP] and cleaved caspase-12) and anti-apoptotic factors (chaperone glucose-regulated protein [GRP] 78 and endoplasmic reticulum-associated protein degradation-associated ubiquitin ligase synovial apoptosis inhibitor 1 [SYVN1]) in the ERS-associated unfolded protein response (UPR) pathway.</p><p><b>RESULTS</b>Compared with age-matched WT mice, 5×FAD mice showed higher cleaved caspase-3, lower neuron-positive staining at the age of 12 months, but earlier cognitive deficit at the age of 7 months (all P < 0.05). Interestingly, for 2-month-old 5×FAD mice, the related proteins involved in the ERS-associated UPR pathway, including CHOP, cleaved caspase-12, GRP 78, and SYVN1, were significantly increased when compared with those in age-matched WT mice (all P < 0.05). Moreover, ERS occurred mainly in neurons, not in astrocytes.</p><p><b>CONCLUSIONS</b>These findings suggest that compared with those of age-matched WT mice, ERS-associated pro-apoptotic and anti-apoptotic proteins are upregulated in 2-month-old 5×FAD mice, consistent with intracellular Aβ aggregation in neurons.</p>