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Aim To observe the effect of thalidomide on the learning and memory ability and hippocampal tissue proteome of Alzheimer's disease(AD)mice,to screen the differential proteins of thalidomide in preventing and treating AD,the pathways involved in regulation,and to explore its possible mechanism. Methods The experimental mice were randomly divided into blank control group,model group,and thalidomide high and low dose groups. The drugs were administered by gavage every day for 21 days. After the administration,Morris water maze test was used to evaluate the learning and memory abilities of the mice,HE staining and Nissl staining were used to observe the pathological tissue morphology of the mouse hippocampus,ELISA was employed to detect the mitochondrial respiratory chain enzyme complex in mouse brain,and the Label-free proteomics method was used to screen different groups of hippocampal proteome proteins. Results The results of the Morris water maze showed that compared with the model group,the escape latency time of the drug group was significantly reduced,and the number of crossing the platform significantly increased(P<0.05). Thalidomide administration could improve the morphological structure of neurons in hippocampus,and could increase the activity of the mitochondrial respiratory chain complex ,Ⅱ, and of the brain tissues of AD mice(P<0.05). A total of 4 378 differential proteins were identified,which had a significant regulatory effect on the expression of 580 proteins in hippocampus of AD mice(P<0.05). Energy metabolism may jointly participate in the regulation of neurodegeneration pathways-changes in pathways such as various diseases and Alzheimer's disease. Conclusions Thalidomide can significantly improve the learning and memory function of AD model mice induced by Aβ
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OBJECTIVES@#To study the changes of protein levels in peripheral blood after it dried.@*METHODS@#The proteins from whole blood and bloodstains were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and normalized by the label-free quantification (LFQ) method. The differential proteins were analyzed by using R 4.2.1 software, limma and edgeR package. The analysis of biological function, signaling pathway and subcellular localization for the differential proteins was then performed.@*RESULTS@#A total of 623 and 596 proteins were detected in whole blood and bloodstains, respectively, of which 31 were statistically significant in the quantitative results, including 10 up-regulated and 21 down-regulated proteins in bloodstains.@*CONCLUSIONS@#The protein abundances in whole blood and bloodstains are highly correlated, and the variation of protein abundances may be related to the changes of endogenous and structural proteins in cells. The application of proteomics technology can assist the screening and identification of protein biomarkers, thereby introducing new biomarkers for forensic research.
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Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Proteomics/methods , Blood Stains , BiomarkersABSTRACT
The present study aimed to explore the mechanism of the sweating of Dipsacus asper on content changes of triterpene sa-ponins by detecting the total triterpene saponins and the index component asperosaponin Ⅵ in the crude and sweated D. asper, and analyzing the differentially expressed proteins by isobaric tags for relative and absolute quantification(iTRAQ) combined with LC-MS/MS. After sweating, the content of total triterpene saponins decreased manifestly, while that of asperosaponin Ⅵ increased significantly. As revealed by the iTRAQ-LC-MS/MS analysis, 140 proteins with significant differential expression were figured out, with 50 up-regulated and 90 down-regulated. GO analysis indicated a variety of hydrolases, oxido-reductases, and transferases in the differential proteins. The results of activity test on two differentially expressed oxido-reductases were consistent with those of the iTRAQ-LC-MS/MS analysis. As demonstrated by the analysis of enzymes related to the triterpene saponin biosynthesis pathway, two enzymes(from CYP450 and UGT families, respectively, which are involved in the structural modification of triterpene saponins) were significantly down-regulated after sweating. The findings suggested that sweating of D. asper presumedly regulated triterpene saponins by affecting the expression of downstream CYP450 s and UGTs in the biosynthesis pathway of triterpene saponins of D. asper.
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Humans , Chromatography, Liquid , Dipsacaceae , Saponins , Sweating , Tandem Mass Spectrometry , TriterpenesABSTRACT
OBJECTIVE@#To screen protein target in prevention and treatment with electroacupuncture (EA) for Alzheimer's disease (AD) and explore the potential mechanism of EA in prevention of AD.@*METHODS@#A total of 40 APP/PS1 transgenic young male mice, 1.5-month old, were randomized into an EA group and a model group, 20 mice in each one, and 20 C57BL/6J mice were chosen as the normal control group. After adaptive housing for 1 week, the mice in the EA group were stimulated with EA at "Baihui" (GV 20), "Fengfu" (GV 16) and "Shenshu" (BL 23), with intermittent wave, 10 Hz in frequency and 2 mA in electric intensity. EA was given once daily, 20 min each time. There was 1 day at interval after EA for 6 days each week. Totally, the intervention lasted for 16 weeks. On day 3 after the end of EA intervention, Morris water maze test was adopted to detect learning and memory abilities of mice in each group. After water maze test, the label-free method was used to measure the difference expressions in cerebral cortex and hippocampus. Using Western blot method, the expressions of guanylate binding protein beta 5 (GNB 5) and histone-H 3 in cerebral cortex and hippocampus were verified. Using immunohistochemical method, the expressions of amyloid beta protein (Aβ) in cerebral cortex and hippocampus were detected.@*RESULTS@#Compared with the normal control group, the escape latency (on day 2, 3 and 4) was prolonged, the frequency of crossing platform and the duration of platform stay were decreased in the mice of the model group (@*CONCLUSION@#The intervention with EA effectively prevents from the decline of learning and memory ability and the formation of Aβ senile plaques in cerebral cortex and hippocampus in young mouse models of AD after growing up. Besides, EA plays a regulatory function for protein expression differences induced by AD model.
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Animals , Male , Mice , Alzheimer Disease/therapy , Amyloid beta-Peptides/genetics , Disease Models, Animal , Electroacupuncture , Hippocampus , Mice, Inbred C57BL , ProteomicsABSTRACT
Objective To investigate the clinical application of Huashi-Xingyu-Qingre decoction for the treatment of oral lichen planus (OLP).Methods According to the random table method,83 OLP patients were divided into control group (n=41) and the research group (n=42).The patients in the control group were treated with conventional medicine,while the patients in the research group were treated with Huashi-Xingyu-Qingre decoction on the basis of control group.Two groups of patients were treated for 6 weeks.The clinical total effective rate of two groups after treatment was counted.The serum haptoglobin,vitamin D binding protein,AT-Ⅲ,IL-6,IL-8,TNF-α and peripheral blood CD3+,CD4+,CD4+/CD8+ were determined respectively before and after treatment.The adverse reaction incidence and recurrence rate of two groups were observed.Results The total effective rate of research group was 92.9％ (39/42),while the total effective rate of control group was 63.4％ (26/41),and the difference between two groups was statistically significant (22=10.589,P=0.001).After treatment,the serum haptoglobin (6.11 ± 0.72 μg/ml vs.5.58 ± 0.69 μg/ml,t=3.423) of research group were significantly higher than the control group (P＜0.05),while the serum vitamin D binding protein (1.48 ± 0.25 μg/ml vs.2.06 ± 0.31 μg/ml,t=9.394),AT-Ⅲ,IL-6,1L-8,TNF-α of research group were significantly lower than the control group (t were 17.561,12.664,7.423,13.763,P＜0.01),and the peripheral blood CD3+,CD4+,CD4+/CD8+ of research group were significantly higher than the control group (t were 5.368,4.694,3.558,P＜0.01).The recurrence rate of control group was 30.8％ (8/26),which was significantly higher than the research group of 7.7％ (3/39) (x2=5.909,P=0.015).Conclusions The clinical curative effect of Huashi-Xingyu-Qingre decoction on the treatment for OLP is significantly,which can improve the patient's serum differential proteins and inflammatory cytokines,improve the body's immune function.It has the high security of the forward curative effect,which provides new thinking for clinical treatment of CFS.
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Objective To analyze the protein abundance differences between two Leishmania infantum strains isolated from different epidemiological types of visceral leishmaniasis in China by comparative proteomics method. Methods Tryptic digests of total proteins were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by label-free quantitative differential expression analysis. The MS data were analyzed with MaxQuant software (ver 1.3.0.5) against data base. Results This study resulted in the identification of 4 274 proteins across two strains (JIASHI-5 and SC6) . Of these, 1 219 differentially expressed proteins (ratio ＞ 2.0 or ＜ 0.5, P < 0.05) were identified. Considering the proteins differentially or uniquely expressed in the strains, 550 proteins were only found in the JIASHI-5 strain, and 174 proteins were only found in the SC6 strain. Totally 495 differentially proteins were expressed in the two groups, among which 328 proteins were down-regulated and 167 proteins were up-regulated in SC6 strain. Some of the identified differentially expressed proteins were demonstrated and they involved in energy metabolism, stress response, prolonging the lifetime of the infected host cell and survival and proliferation in virulent strains. Conclusion This study reveals a group of differentially expressed proteins and the related biologic function that may lay the foundation for screening and identification of the key Leishmania molecules relative to pathogenicity.
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Objective To analyze the protein abundance differences between two Leishmania infantum strains isolated from different epidemiological types of visceral leishmaniasis in China by comparative proteomics method. Methods Tryptic digests of total proteins were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by label-free quantitative differential expression analysis. The MS data were analyzed with MaxQuant software (ver 1.3.0.5) against data base. Results This study resulted in the identification of 4 274 proteins across two strains (JIASHI-5 and SC6) . Of these, 1 219 differentially expressed proteins (ratio ＞ 2.0 or ＜ 0.5, P < 0.05) were identified. Considering the proteins differentially or uniquely expressed in the strains, 550 proteins were only found in the JIASHI-5 strain, and 174 proteins were only found in the SC6 strain. Totally 495 differentially proteins were expressed in the two groups, among which 328 proteins were down-regulated and 167 proteins were up-regulated in SC6 strain. Some of the identified differentially expressed proteins were demonstrated and they involved in energy metabolism, stress response, prolonging the lifetime of the infected host cell and survival and proliferation in virulent strains. Conclusion This study reveals a group of differentially expressed proteins and the related biologic function that may lay the foundation for screening and identification of the key Leishmania molecules relative to pathogenicity.
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This article was aimed to study the proteomic spectra expression of traditional Chinese medicine (TCM) cold and heat constitution rats with two-dimensional gel electrophoresis (2-DE), in order to search for the cold and heat-associated proteins for the investigation of the biological basis of TCM cold and heat body constitution formation. The total protein in rat’s liver cell was extracted. The 2-DE and MALDI-TOF/TOF mass spectrometry (MS) were used in the screening and identification of differentially expressed proteins of cold and heat constitution rats. The results showed that a total of 10 different points in the protein expression were obtained with statistical significance after screening and MS, which were carbamoyl-phosphate synthase, protein disulfide isomerase associated 3, catalase, hydroperoxide isomerase, cytosol aminopeptidase, glutamate dehydrogenase 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, heat shock protein 60 (Hsp60) precursor, homocysteine, 78 kDa glucose-regulated protein precursor. It was concluded that some differences were existed in the proteomic spectra expression of TCM cold and heat constitution rats. The abnormality of enzyme protein metabolism may be one of the material bases for the formation of cold and heat constitution.
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Objective To obtain the peptide mass fingerprintings(PMF)of serums of the patients with bone metastasis and with-out metastasis and to filtrate and identify the differential serum proteins of patients with bone metastasis after radical mastectomy . To establish diagnostic models for diagnosis of bone metastasis after breast cancer operation .Methods Two groups of serum sam-ples were analyzed by ClinprotTM MALDI-TOF MS and gain PMF ,18 samples from patients with merely bone metastasis and 18 samples from patients without metastasis .Characteristic protein peaks were analyzed and selected by analyses software within Clin-prot system .Every group was repeated 2 times .Results All serum samples were repeated after 5 days and the fingerprintings were similar to the former .4 protein peaks were selected randomly to compute coefficient of variation which are less than 30% .The Clin-prot system has excellent repeatability .No differential protein was detected by analyzing PMF (P>0 .05) .Conclusion No differen-tial serum protein exists in patients with bone metastasis and without metastasis and detecting differential protein in peripheral blood may be impossible .
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Aim To identify paclitaxel differential binding protein in the cells of cell line MCF-7 and MCF-7/Taxol and to find new target for antitumor agents.Methods Synthesis and activity assay of biotinylated paclitaxel was used to gain paclitaxel binding protein by semiautomatic in vitro selection using the affinity magnetic beads method.LC MS/MS analysis and Western immunoblot analysis were used to identify the differential binding protein.Results The experimentation identified paclitaxel binding protein in the cells of MCF-7 and MCF-7/Taxol.An absent strap in MCF-7/Taxol was discovered by comparing the straps of two cell lines, which contained 25 kinds of proteins, among which 3 proteins were identified by western blot techniques: Heat shock protein HSP 90,Dermcidin Precursor,Actinin.Conclusions By comparing the straps of two cell lines, the differential protein in the cells of MCF-7 and MCF-7/Taxol are discovered, implying that they may be the novel mechanism of taxane resistance and may lead to find a new approach to finding a new target for oncotherapy drugs.
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AIM:To find new biomarkers in the sera of retinoblastoma (Rb) patients with surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI TOF MS) and protein chip technique.METHODS:SELDI TOF MS, IMAC30 and CM10 protein chips were used to analyze the protein profiles from sera of 18 patients with Rb and 17 age matched controls. The protein profiling was analyzed statistically by Ciphergen protein chip software 3.0.2. The test was applied to compare the protein peak intensity. Fisher's exact test was used to compare the predominance of differential protein peaks appeared in patients.RESULTS:With IMAC30 protein chips, there were 26 proteins which appeared different in sera of patients with Rb compared to normal children. Among them, 21 proteins, I.e. 7746, 7014, 11713, 3049, 7084, 7299, 5888, 2544, 12575, 5489, 9658, 9575, 9929, 10161, 8955, 1886, 10617, 6209, 2411, 7374, 6614m/z were up regulated and 5 proteins, I.e. 8382, 7923, 7972, 8590, 66576m/z, were down regulated(P<0.01). Using the 7014 protein peak for statistical analysis, we could differentiate the patients with Rb from the healthy children with a sensitivity of 94.4% and a specificity of 82.4%. By CM10 protein chips, 4 proteins, including 3 up regulated proteins(5888, 6097, 7798 and 1 down regulated protein (8590m/z), were detected in Rb patients (P<0.01). The sensitivity and specificity were 83.3% and 70.6% respectively when 7798m/z protein peak was selected for statistical analysis.CONCLUSION:There are a few candidates as Rb biomarkers in the sera of Rb patients. SELDI TOF MS protein chip technology could be a potential method in the clinical screening test of Rb.
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Objective To explore primarily the differentially expressed proteins in the hemolymph from adult female Anopheles stephensi ( An stephensi ) infected with Plasmodium yoelii ( P yoelii ) after being fed with sucrose solution containing nitroquine or not at different time points Methods Hemolymph of 2 groups of adult female An stephensi was collected with the expulsion method from the first day to the fifth day after the feeding Hemolymph samples were examined with SDS PAGE The protein gels were visualized by either Coomassie brilliant blue or silver staining, scanned and automatically analyzed by the BioRad1000 gel image analysis system for differential proteins bands Results On the second day of feeding with nitroquine, a few oocysts were partially melanized Furthermore, during the period from the fifth day to the ninth day, the number of mosquitoes with malanized oocysts and the number of melanized oocysts gradually increased The number of hemolymph protein binds in the treatment group was markedly more than that in the control Many different bands, mainly located at the molecular weight of (20～40)?10 3 and (60～80)?10 3, were visualized in the 2 groups The number of protein bands stained by the silver staining was more than that by the Coomassie brilliant blue staining Conclusion There are differentially expressed proteins in the hemolymph in An stephensi infected with P yoelii after being fed with sucrose solution containing nitroquine These differential proteins may be the melanization engaging proteins
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Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.