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ObjectiveHenoch-Schönlein purpura（HSP） is one of the dominant diseases in Mongolian medicine. Qishun Baolier（QSBLE）， as the main prescription for the treatment of HSP， has significant clinical effect， but its mechanism is not yet clear. Baed on this， this study is intended to screen the differentially expressed proteins before and after treatment， and preliminarily explore the molecular mechanism of QSBLE in the treatment of HSP. MethodTaking oneself as the control， 30 HSP patients aged 6-45 years were collected， and QSBLE was taken orally at 12：00 and 24：00， respectively. The dose was adjusted according to age and the course of treatment was one week. The distribution of proteinuria， hematuria and skin purpura of all patients were determined before and after treatment. The serum samples of 10 patients with clinically significant remission after QSBLE treatment were randomly selected for proteomics. Isobaric tags for relative and absolute quantification（iTRAQ） combined with liquid chromatography tandem mass spectrometry（LC-MS/MS） was used to analyze the proteins in serum of HSP patients before and after treatment， and differential proteins were analyzed bioinformatically and the protein-protein interaction（PPI） networks were constructed. ResultA total of 378 proteins were identified from serum， including 18 differentially expressed proteins， of which 15 proteins were up-regulated and 3 proteins were down regulated. Bioinformatics showed that the differential proteins were mainly involved in biological processes such as immune response， immunoglobulin production， phagocytosis， adaptive immune response before and after treatment. Biological processes， pathways and proteins were used to construct the PPI network， the proteins represented by immunoglobulin heavy constant γ1（IGHG1）， immunoglobulin λ-chain 7-43（IGLV7-43）， gelsolin（GSN） and 60 kDa heat shock protein（HSPD1） were involved in biological processes and related pathways such as adaptive immune response， immunoglobulin production， leukocyte-mediated immunity， regulation of stress response， regulation of immune system processes， regulation of trauma response， and these proteins were at the center of the PPI network. ConclusionQSBLE may play a role in the treatment of HSP by regulating the expression of IGHG1， IGLV7-43， GSN， HSPD1 and other key proteins to affect immune-related biological processes.
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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.
Subject(s)
Humans , Chromatography, Liquid , Dipsacaceae , Saponins , Sweating , Tandem Mass Spectrometry , TriterpenesABSTRACT
Objective To compare the differentially expressed proteins between cypermethrin-resistant and -sensitive Culex pipiens pallens, so as to unravel the mechanism underlying the resistance to cypermethrin in Cx. p. pallens. Methods A quantitative proteomic analysis was performed among cypermethrin-sensitive and -resistant isolates of Cx. p. pallens using isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results A total of 164 differentially expressed proteins were identified between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, including 54 up-regulated proteins and 110 down-regulated proteins. A large number of cuticular proteins, larval cuticular proteins, pupal cuticular proteins and cuticular structural constituent proteins, which are associated with cytoskeletal structure and components, were differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens. Thirteen proteins, which were involved in energy production and conversion, translation, ribosomal structure and biogenesis, lipid transport and metabolism, post-translational modification, protein turnover, chaperones, cytoskeleton and intracellular transportation, were validated to be differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, which may serve as potential markers of cypermethrin resistance. Conclusion Multiple insecticide resistance mechanisms contribute to the resistance to cypermethrin in Cx. p. pallens, including cuticular resistance and metabolic resistance, and the cuticular protein genes and cytochrome P450 enzymes may play an important role in the resistance of Cx. p. pallens to cypermethrin.
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In the past decade, relative proteomic quantification using isobaric labeling technology has developed into a key tool for comparing the expression of proteins in biological samples. Although its multiplexing capacity and flexibility make this a valuable technology for addressing various biological questions, its quantitative accuracy and precision still pose significant challenges to the reliability of its quantification results. Here, we give a detailed overview of the different kinds of isobaric mass tags and the advantages and disadvantages of the isobaric labeling method. We also discuss which precautions should be taken at each step of the isobaric labeling workflow, to obtain reliable quantification results in large-scale quantitative proteomics experiments. In the last section, we discuss the broad applications of the isobaric labeling technology in biological and clinical studies, with an emphasis on thermal proteome profiling and proteogenomics.
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Proteome/metabolism , Proteomics/methods , Reproducibility of Results , Tandem Mass Spectrometry/methodsABSTRACT
Objective To compare the difference of protein expression between the post-overwintering stage and the diapauses preparation stage in Culex pipiens pallens, so as to reveal the mechanisms underlying the overwintering diapause of Cx. pipienspallens. Methods A quantitative proteomic analysis was performed in Cx. pipiens pallens before and after overwintering diapause by using isobaric tags for relative and absolute quantification (iTRAQ) labeling. Results A total of 244 differentially expressed proteins were identified in Cx. pipiens pallens before and after overwintering diapause, including 126 up-regulated proteins and 118 down-regulated proteins. iTRAQ-based quantitative proteomic analysis revealed that these differentially expressed proteins were linked to function and energy production and conversion, lipid metabolism, remodeling of cytoskeleton, carbohydrate metabolism, protein transport, molecular chaperones, stress tolerance and metabolic enzymes. Conclusions This is the first study to identify the overwintering diapause-related proteins in Cx. pipiens pallens using proteomics tools, which reveals KEGG pathways and GO terms associated with the overwintering diapauses of Cx. pipiens pallens. Our findings provide additional understandings pertaining to the mechanisms underlying the overwintering diapauses of Cx. pipiens pallens.
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Objective To compare the difference of protein expression between the post-overwintering stage and the diapauses preparation stage in Culex pipiens pallens, so as to reveal the mechanisms underlying the overwintering diapause of Cx. pipienspallens. Methods A quantitative proteomic analysis was performed in Cx. pipiens pallens before and after overwintering diapause by using isobaric tags for relative and absolute quantification (iTRAQ) labeling. Results A total of 244 differentially expressed proteins were identified in Cx. pipiens pallens before and after overwintering diapause, including 126 up-regulated proteins and 118 down-regulated proteins. iTRAQ-based quantitative proteomic analysis revealed that these differentially expressed proteins were linked to function and energy production and conversion, lipid metabolism, remodeling of cytoskeleton, carbohydrate metabolism, protein transport, molecular chaperones, stress tolerance and metabolic enzymes. Conclusions This is the first study to identify the overwintering diapause-related proteins in Cx. pipiens pallens using proteomics tools, which reveals KEGG pathways and GO terms associated with the overwintering diapauses of Cx. pipiens pallens. Our findings provide additional understandings pertaining to the mechanisms underlying the overwintering diapauses of Cx. pipiens pallens.
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Aim: To compare the differential expression of proteins in testicular tissues of ionized irradiated mice and normal mice, in order to explain the mechanism of ionizing irradiation on testicular tissue damage from proteomics perspective. Methods: The mouse ionizing radiation damage model was established by cobalt 60-ray irradiation. Comparative proteomics method, iTRAQ combined with LC-MS/MS detection technology were used to extract differentially expressed proteins of testis tissues from mouse irradiated group and normal group, then David6. 8, StringlO. 5 and Cytoscape 3. 6. 1 database were employed for KEGG enrichment analysis and interaction analysis on differential proteins. Results: Twenty-one biological signaling pathways (P <0. 05) were identified by KEGG enrichment analysis, with 13 differentially expressed proteins enriched in phosphoric oxide signaling pathways, and both were downgrades of expression. The pathway involved several subgroups of ATP synthase, cytochrome and NADH dehydrogenase, which were mainly involved in mitochondrial electron transfer, mitochondrial respiratory chain complex I assembly, ATP biosynthesis and so on. Conclusions: Ionizing radiation is responsible for the expression of oxidative phosphorylation signaling pathway in mouse testis tissues. The low expression of 13 differential proteins leads to the synthesis of cell ATP. It is an important mechanism of radiation damage.
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Objective@#Hepatocellular carcinoma (HCC) is one of the most common malignant tumor worldwide. Metastasis is a marker of cancer deterioration in patients with liver cancer and a major cause of death. In order to develop effective therapeutic strategies, it is urgent to study the molecular basis of liver cancer metastasis.@*Methods@#Immunohistochemistry was used to detect the expression of fatty acid synthase (FASN) in HCC. Wound healing and transwell cell invasion assays was used to confirm the role of FASN in liver cancer migration and invasion. Proteins that interacted with FASN were identified using iTRAQ (isobaric tag for relative and absolute quantification). Co-immunoprecipitation (Co-IP) and cellular immunofluorescence analysis were used to assess the interaction between FASN and signal transduction and transcription activator 3 (STAT3). The expression of STAT3, p-STAT3, matrix metalloproteinase (MMP)-2 and MMP-9 was detected after FASN knockdown using Western blot method. Statistical analysis was performed using the t-test.@*Results@#Immunohistochemistry showed that the expression of FASN in HCC tissue was higher than that in adjacent tissues. iTRAQ, Co-IP and immunofluorescence analysis revealed that FASN interacted with STAT3. Western blot analysis showed that the expression of p-STAT3, MMP-2 and MMP-9 decreased after FASN knockdown.@*Conclusion@#FASN may promote the metastasis of liver cancer by interacting with STAT3 and affecting the expression of MMP-2/MMP-9.
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Aim To clarify the damage mechanisms of ionizing radiation on mouse spleen tissues and analyse differential protein expression in spleen tissues of mice exposed to ionizing radiation injury and normal mice from the perspective of comparative proteomics, in order to analyse the signaling pathways involved in differential proteins. Methods The mouse ionizing radiation injury model was established by 5 Gy 60Co-y ray, and the mouse spleen tissue was extracted in the ionizing radiation group and normal group, then ITRAQ combined with LC-MS/MS detection technology were used to screen differentially expressed proteins of spleen tissues from ionizing radiation group and normal group. Results A total of 17 biological signaling pathways were identified by KEGG pathway enrichment analysis(P <0. 0 5) , in which 37 differential expressed proteins were enriched in the ribosome signaling pathway pathway , including 36 differential expressed proteins downregulated and one differential expressed protein up-regulated. These pathway involved several multiple subunit proteins, such as ribosome proteins, 60S ribosome proteins and 40S ribosome proteins, which were mainly involved in biological processes, such as translation of proteins, structural composition of ribosome and so on. Conclusions Ionizing radiation causes 37 differential proteins involved in ribosome signaling pathway of mouse spleen tissues, including 36 differential expressed proteins down-regulated and one differential expressed protein up-regulated. The disturbance of protein synthesis in spleen tissues is an important mechanism of ionizing radiation injury.
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The vegetative state is a complex condition with unclear mechanisms and limited diagnostic, prognostic, and therapeutic methods. In this study, we aimed to explore the proteomic profile of tears from patients in a traumatic vegetative state and identify potential diagnostic markers using tears-a body fluid that can be collected non-invasively. Using iTRAQ quantitative proteomic technology, in the discovery phase, tear samples collected from 16 patients in a traumatic vegetative state and 16 normal individuals were analyzed. Among 1080 identified tear proteins, 57 were upregulated and 15 were downregulated in the patients compared to the controls. Bioinformatics analysis revealed that the differentially-expressed proteins were mainly involved in the wound response and immune response signaling pathways. Furthermore, we verified the levels of 7 differentially-expressed proteins in tears from 50 traumatic vegetative state patients and 50 normal controls (including the samples used in the discovery phase) using ELISA. The results showed that this 7-protein panel had a high discrimination ability for traumatic vegetative state (area under the curve = 0.999). In summary, the altered tear proteomic profile identified in this study provides a basis for potential tear protein markers for diagnosis and prognosis of the traumatic vegetative state and also provides novel insights into the mechanisms of traumatic vegetative state.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Biomarkers , Metabolism , Chromatography, Liquid , Enzyme-Linked Immunosorbent Assay , Eye Proteins , Metabolism , Mass Spectrometry , Persistent Vegetative State , Metabolism , Proteome , Proteomics , ROC Curve , Tears , MetabolismABSTRACT
Objective We performed comprehensive proteomic analyses of synovial fluid by using the isobaric tags for relative and absolute quantitation (iTRAQ) method and LC-MS/MS, and searched for candidate biomarkers for osteoarthritis. Methods Synovial fluid was collected from patients with late-stage OA, earlystage OA and the control group. Molecular variations were detected by the iTRAQ method. T test was used for statistical analysis. Results Using the iTRAQ method, we identified 1 283 proteins from synovial fluid of patients with osteoarthritis, of which 268 proteins were not reported previously. There were 72 proteins upregulated and 249 proteins down-regulated between moderate OA group and controls. One hundred and twentyeight proteins were up-regulated and 141 proteins were down-regulated between severe OA group and controls.One hundred and ninety-two proteins were up-regulated and 76 proteins were down-regulated between severe OA group and moderate OA group. Eight proteins were found to be up-regulated in the three groups.Conclusion This is an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis by iTRAQ method. The catalog of proteins generated in this study will further expand our knowledge regarding the pathophysiology of osteoarthritis and help us in identifying good biomarkers for early diagnosis.
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Objective We performed comprehensive proteomic analyses of synovial fluid by using the isobaric tags for relative and absolute quantitation (iTRAQ) method and LC-MS/MS, and searched for candidate biomarkers for osteoarthritis. Methods Synovial fluid was collected from patients with late-stage OA, earlystage OA and the control group. Molecular variations were detected by the iTRAQ method. T test was used for statistical analysis. Results Using the iTRAQ method, we identified 1 283 proteins from synovial fluid of patients with osteoarthritis, of which 268 proteins were not reported previously. There were 72 proteins upregulated and 249 proteins down-regulated between moderate OA group and controls. One hundred and twentyeight proteins were up-regulated and 141 proteins were down-regulated between severe OA group and controls.One hundred and ninety-two proteins were up-regulated and 76 proteins were down-regulated between severe OA group and moderate OA group. Eight proteins were found to be up-regulated in the three groups.Conclusion This is an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis by iTRAQ method. The catalog of proteins generated in this study will further expand our knowledge regarding the pathophysiology of osteoarthritis and help us in identifying good biomarkers for early diagnosis.
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Objective To investigate the clinical value of iTRAQ multiplex tandem mass spectrometry in the detection of the ex-pression of hepatocyte growth factor (HGF) in patients with invasive breast cancer .Methods A total of 35 patients with breast cancer and 30 healthy subjects were selected from January 2014 to October 2016 in this hospital ,the expression of serum HGF in breast cancer patients with different clinical stages and healthy subjects was analyzed by iTRAQ labeling ,mass spectrometry ,library searching and Scqffold software ,and the differential expression of HGF was verified by Western blot .Results A total of 237 pro-teins were identified in the serum samples of this study ,and 89 proteins with strict quantitative criteria ,17 differentially expressed proteins ,included HGF ,were screened for breast cancer patients and healthy controls .iTRAQ markers showed that the expression level of serum HGF in different clinical stage of breast cancer patients was significantly higher than that in healthy subjects (P<0 .05) .The results of Western blot showed that the relative expression level of serum HGF in breast cancer patients was significant-ly higher than that in healthy subjects(P<0 .05) .Conclusion iTRAQ multiplex tandem mass spectrometry is useful for the detection of breast cancer patients with high expression of HGF ,which is of great significance in guiding the clinical treatment of breast cancer .
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Objective To analyze the protein expression profile of HeLa cells transfected with pORF5 gene of Chlamydia trachomatis. Methods A lentiviral expression vector containing pORF5 gene was constructed. The lentiviral expression vector and helper plasmids were co-transfected into 293T cells to construct the recombinant lentivirus, which was used to infect HeLa cells. HeLa cells transfected with pORF5 gene and control HeLa cells were sorted out by flow cytometry. The isobaric tags for relative and absolute quantitation ( iTRAQ) approach combined with nano-liquid chromatography-tandem mass spec-trometry ( NanoLC-MS/MS) analysis was performed to understand protein expression profiles and to iden-tify and quantify the differentially expressed proteins in the pORF5-transfected HeLa cells ( pORF5-Hela) and the control HeLa cells. Quantitative real-time PCR ( qRT-PCR ) and Western blot analysis were performed to detect the expression of some proteins at mRNA and protein levels, respectively. Results HeLa cell line stably transfected with pORF5 gene and control HeLa cell line were constructed successful-ly. Totally 314 proteins were differentially expressed between the pORF5-HeLa and control HeLa cells, 159 of which showed increased expression and the other 155 showed decreased expression in pORF5-HeLa cells. The differentially expressed proteins were involved in many processes, such as metabolic process, immune response, biological adhesion and so on. Results of qRT-PCR showed that the expression of HIST1H1C(histone H1. 2C), HBA1(hemoglobin subunit alpha), PARK7(parkinson disease protein 7), HMGB1(high mobility group protein B1) and HMGB2 at mRNA level in pORF5-HeLa cells were up-regulated, while the expression of CLIC1 ( chloride intracellular channel protein 1 ) , KRT7 ( typeⅡ cy-toskeletal 7), SFN(14-3-3 protein sigma) and CDKN2A(cyclin-dependent kinase inhibitor 2A) were down-regulated. Western blot analysis confirmed the enhanced expression of HMGB1 and PRAK7 at pro-tein level. The results of qRT-PCR and Western blot analysis were consistent with proteomic data. Con-clusion Expression profiles for differentially expressed proteins between pORF5-HeLa and control HeLa cells were established successfully. The differentially expressed proteins regulated by pORF5 gene were found to be related to cell metabolism, proliferation, adhesion and so on, suggesting that pORF5 might promote the growth and proliferation of Ct by regulating protein expression and biological behavior of host cells.
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Objective To investigate the clinical value of iTRAQ multiplex tandem mass spectrometry in the detection of the ex-pression of hepatocyte growth factor (HGF) in patients with invasive breast cancer .Methods A total of 35 patients with breast cancer and 30 healthy subjects were selected from January 2014 to October 2016 in this hospital ,the expression of serum HGF in breast cancer patients with different clinical stages and healthy subjects was analyzed by iTRAQ labeling ,mass spectrometry ,library searching and Scqffold software ,and the differential expression of HGF was verified by Western blot .Results A total of 237 pro-teins were identified in the serum samples of this study ,and 89 proteins with strict quantitative criteria ,17 differentially expressed proteins ,included HGF ,were screened for breast cancer patients and healthy controls .iTRAQ markers showed that the expression level of serum HGF in different clinical stage of breast cancer patients was significantly higher than that in healthy subjects (P<0 .05) .The results of Western blot showed that the relative expression level of serum HGF in breast cancer patients was significant-ly higher than that in healthy subjects(P<0 .05) .Conclusion iTRAQ multiplex tandem mass spectrometry is useful for the detection of breast cancer patients with high expression of HGF ,which is of great significance in guiding the clinical treatment of breast cancer .
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Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V⁺) and uninfected (V⁻) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V⁺ compared with V⁻ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V⁺ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V⁺ and V⁻ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.
Subject(s)
Glucose-6-Phosphate Isomerase , Glycogen Phosphorylase , Heat-Shock Proteins , Host-Parasite Interactions , Malate Dehydrogenase , Metabolism , Polymerase Chain Reaction , Proteome , Reticuloendotheliosis virus , Ribosomal Proteins , RNA, Double-Stranded , RNA, Messenger , Trichomonas vaginalis , Trichomonas , Triose-Phosphate Isomerase , VirulenceABSTRACT
This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using iTRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by iTRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO (gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories:heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; metE and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.
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To explore the associated proteins of the hypothalamus in aging rat models with intervention by Qiongyugao(QYG) based on iTRAQ technology, find out the target protein candidates and investigate the mechanism of delaying aging for Qiongyugao. The results showed that Qiongyugao increased GSH-Px activity in serum and SOD activity in liver; the total protein count identified by iTRAQ was 3 522, FDR<1%. There were 20 kinds of differential proteins between the blank group and model group; there were 295 kinds of differential proteins between model group and QYG group, and 40 kinds of them had a difference multiple ≥1.30 (the maximum value was 1.47). Compared with blank group, there were 14 kinds of proteins that were down-regulated in model group and up-regulated in QYG group. Combined with literature search and gene function search, 12 kinds of target protein candidates were screened out ： ST18, Ptprc, PSMB8, INPP4B, Shc3, Pik3r1, PIP5K1C, Nampt, Rasgrp2, Asah2, Pdpk1, and Map2k7. The expression of nuclear factor-kappa B (NF-κB) in the hypothalamic inflammatory pathway was detected by Western blot and the results showed that its expression level in model group(0.96) was higher than that in control group(0.85), while its expression level in QYG group(0.89) was lower than that in model group. Q-PCR results showed that the relative mRNA expression levels of PIP5K1C and Ptprc in model group were significantly lower than those in blank group(P<0.01); while compared with the model group, the mRNA expression levels of PIP5K1C and Ptprc in QYG group were significantly increased(P<0.01) . This result was consistent with proteomics data. QYG may delay aging by regulating hypothalamic inflammatory reaction.
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Coptidis Rhizoma is commonly used in clinical medicine. It has been widely used in traditional Chinese medicine (TCM), with the functions of clearing heat, drying dampness, purging intense heat relieve toxins. However, the herb contains complex ingredients, and CYP450 isoenzyme is the main metabolic enzyme of the drug, so it is of important clinical significance to study the effect of Coptidis Rhizoma on cytochrome P450 enzymes. In the experiment, liver tissues of rats were selected and liver microsomes were separated by differential centrifugation. Bicinchoninic acid (BCA) Protein quantitation was done by enzyme linked immunosorbent assay, and iTRAQ (isobaric tags for relative and absolute quantification) was combined with the 2D-LC-MS/MS analysis method for identification of CYP450 proteins. With Coptidis Rhizoma, 30 CYP450 isoenzymes were identified, with 7 proteins significantly increased and 8 decreased in response to Coptidis Rhizoma, while the rest 15 had no change. iTRAQ technology combined with 2D-LC-MS/MS method could be used to comprehensively study CYP450 enzyme, but it is necessary to further evaluate in vitro levels of Coptidis Rhizoma and avoid any potential clinical drug-drug interaction.
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Aims To compare hippocampus between CUMS rats and normal rats, to find differentially ex-pressed proteins and to explore the pathogenesis of de-pression in the protein levels and biological marker. Methods Chronic unpredicted mild stress was taken to establish rat depression model. The ITRAQ-labeled proteins and peptides were separated by the cation col-umn, and differentially expressed proteins were detec-ted and identified by 2D LC-MS/MS. The functions of proteins were analyzed by bioinformatics. Results To-tally 5 109 proteins were identified, 33 differentially expressed proteins were identified, the expressions of 8 proteins were increased and 25 proteins downregulated. Conclusion ITRAQ based sereening is effective in discovering the nosogenesis of depression and new bio-logical marker.