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proteomic analysis reveals distinctive protein profiles involved in CD8^(+) T cell-mediated murine autoimmune cholangitis

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Cellular & Molecular Immunology 2018年第8期15卷 756-767页

作者： Weici Zhang Ren Zhang Jun Zhang Ying Sun Patrick SC Leung Guo-Xiang Yang Zongwen Shuai William M Ridgway M Eric Gershwin Division of Rheumatology Allergy and Clinical ImmunologyUniversity of CaliforniaDavisCaliforniaUSA Department of Pathogenic Biology and Immunology Guangzhou University of Chinese MedicineGuangzhou 510006China Center for non-infectious liver diseases Beijing 302 HospitalBeijing 100039China Department of Rheumatology and Immunology The First Affiliated Hospital of Anhui Medical UniversityHefei 230022China Division of Immunology Allergy and RheumatologyUniversity of CincinnatiCincinnatiOH 45267

Autoimmune cholangitis arises from abnormal innate and adaptive immune responses in the liver,and T cells are critical drivers in this process.However,little is known about the regulation of their functional behavior during disease development.We previously reported that mice with T cell-restricted expression of a dominant negative form of transforming growth factor beta receptor type Ⅱ(dnTGFβ RII)spontaneously develop an autoimmune cholangitis that resembles human primary biliary cholangitis(PBC).Adoptive transfer of CD8^(+) but not CD4^(+) T cells into Rag1^(−/−)mice reproduced the disease,demonstrating a critical role for CD8^(+) T cells in PBC pathogenesis.Herein,we used SOMAscan technology to perform proteomic analysis of serum samples from dnTGFβRII and B6 control mice at different ages.In addition,we analyzed CD8 protein profiles after adoptive transfer of splenic CD8^(+) cells into Rag1^(−/−)recipients.The use of the unique SOMAscan aptamer technology revealed critical and distinct profiles of CD8 cells,which are key to biliary mediation.In total,254 proteins were significantly increased while 216 proteins were significantly decreased in recipient hepatic CD8^(+) cells compared to donor splenic CD8^(+) cells.In contrast to donor splenic CD8^(+) cells,recipient hepatic CD8^(+) cells expressed distinct profiles for proteins involved in chemokine signaling,focal adhesion,T cell receptor and natural killer cell-mediated cytotoxicity pathways.

关键词： autoimmune cholangitis CD8 dnTGFβRII mice proteomic analysis

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proteomic analysis of a chronic obstructive pulmonary disease mouse model to determine the efficacy of treatment using Guben Zhike decoction

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Journal of Traditional Chinese Medical Sciences 2021年第1期8卷 34-42页

作者： Mingzhe Wang Guoxing Liu Yao Xiao Zhe Cai Chang Liu Lin Pan Ying Liu Mengchao Liu Hongchun Zhang Graduate School Beijing University of Chinese MedicineBeijing 100029China National Clinical Research Center for Respiratory Diseases and Traditional Chinese Medicine Department of Pulmonary Diseases China-Japan Friendship HospitalBeijing 100029China Respiratory Department Beijing University of Chinese Medicine Third Affiliated HospitalBeijing 100029China Nephropathy Department Dongzhimen Hospital Beijing University of Chinese MedicineBeijing 100700China Clinical Research Institute China-Japan Friendship HospitalBeijing 100029China No.4 Department of Healthcare China-Japan Friendship HospitalBeijing 100029China Nephropathy Department Beijing Traditional Chinese Medical HospitalBeijing 100010China

Background:Guben Zhike decoction(GBZKD)is derived from the experience of Professor Enxiang Chao,an esteemed master of Chinese medicine,while treating chronic obstructive pulmonary disease(COPD).GBZKD reinforces the healthy qi and consolidates defensive qi.This study explored the efficacy and potential mechanism of action of GBZKD in a COPD mouse model using proteomics.Methods:A COPD mouse model was established through cigarette smoke exposure and intranasal lipopolysaccharide administration.The model was verified through lung function test and lung histopathological observation.Label-free quantitative proteomics was used to detect the lung tissue proteins of mice from the GBZKD,COPD,and control groups.Results:GBZKD markedly improved the lung function and associated pathological conditions in the COPD mouse model.proteomic analysis identified 4316 proteins,of which 3696 were quantitative proteins.We highlighted 287 and 184 proteins with significant regulatory roles in the lung tissues of COPD mice and GBZKD-treated mice,respectively.These proteins participated in multiple functions,including complement/coagulation cascade,immune response,and metabolic pathways.Conclusion:GBZKD exhibits multitarget and multipathway therapeutic effects in a COPD mouse model.

关键词： Guben zhike decoction Chronic obstructive pulmonary disease proteomic analysis Mouse model

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A highly efficient protein corona-based proteomic analysis strategy for the discovery of pharmacodynamic biomarkers

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Journal of Pharmaceutical analysis 2022年第6期12卷 879-888页

作者： Yuqing Meng Jiayun Chen Yanqing Liu Yongping Zhu Yin-Kwan Wong Haining Lyu Qiaoli Shi Fei Xia Liwei Gu Xinwei Zhang Peng Gao Huan Tang Qiuyan Guo Chong Qiu Chengchao Xu Xiao He Junzhe Zhang Jigang Wang Artemisinin Research Center and Institute of Chinese Materia Medica China Academy of Chinese Medical SciencesBeijing100700China Department of Biological Sciences National University of Singapore117543Singapore Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety Institute of High Energy PhysicsChinese Academy of SciencesBeijing100049China

The composition of serum is extremely complex,which complicates the discovery of new pharmacodynamic biomarkers via serum proteome for disease prediction and diagnosis.Recently,nanoparticles have been reported to efficiently reduce the proportion of high-abundance proteins and enrich lowabundance proteins in serum.Here,we synthesized a silica-coated iron oxide nanoparticle and developed a highly efficient and reproducible protein corona(PC)-based proteomic analysis strategy to improve the range of serum proteomic analysis.We identified 1,070 proteins with a median coefficient of variation of 12.56%using PC-based proteomic analysis,which was twice the number of proteins identified by direct digestion.There were also more biological processes enriched with these proteins.We applied this strategy to identify more pharmacodynamic biomarkers on collagen-induced arthritis(CIA)rat model treated with methotrexate(MTX).The bioinformatic results indicated that 485 differentially expressed proteins(DEPs)were found in CIA rats,of which 323 DEPs recovered to near normal levels after treatment with MTX.This strategy can not only help enhance our understanding of the mechanisms of disease and drug action through serum proteomics studies,but also provide more pharmacodynamic biomarkers for disease prediction,diagnosis,and treatment.

关键词： Protein corona Nanoparticles Mass spectrometry proteomic analysis Pharmacodynamic biomarkers

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proteomic analysis of energy metabolism and signal transduction in irradiated melanoma cells

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International Journal of Ophthalmology(English edition) 2013年第3期6卷 286-294页

作者： Lu-Bin Yan Kai Shi Zhi-Tong Bing Yi-Lan Sun Yang Shen Department of Surgery the Second Hospital of Lanzhou University Department of Ophthalmology the Second Hospital of Lanzhou University Institute of Modern Physics Chinese Academy of Sciences

AIM: To analyze proteomic and signal transduction alterations in irradiated melanoma cells. METHODS: We combined stable isotope labeling with amino acids in cell culture (SILAC) with highly sensitive shotgun tandem mass spectrometry (MS) to create an efficient approach for protein quantification. Protein protein interaction was used to analyze relationships among proteins. RESULTS: Energy metabolism protein levels were significantly different in glycolysis and not significantly different in oxidative phosphorylation after irradiation. Conversely, tumor suppressor proteins related to cell growth and development were downregulated, and those related to cell death and cell cycle were upregulated in irradiated cells. CONCLUSION: Our results indicate that irradiation induces differential expression of the 29 identified proteins closely related to cell survival, cell cycle arrest, and growth inhibition. The data may provide new insights into the pathogenesis of uveal melanoma and guide appropriate radiotherapy.

关键词： melanoma cell 2D-LC-MS/MS stable isotope labeling with amino acids proteomic analysis X-ray irradiation protein-protein interaction

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proteomic analysis of differentially expressed proteins between Xiangyou 15 variety and the mutant M15

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Frontiers in Biology 2014年第3期9卷 234-243页

作者： Zhen-Qian ZHANG Gang XIAO Rui-Yang LIU Tai-Long TAN Chun-Yun GUAN Guo-Huai WANG She-Yuan CHEN Xian-Meng WU Mei GUAN Qin LI Hunan Branch of National Oilseed Crops Improvement Centre Hunan Agricultural University Changsha Hunan 41 O128 China Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization Changsha Hunan 410128 China

A high oleic acid rapeseed material MI5 （derived from Xiangyou 15 variety） has been received more attention for its significant effect for human health. And it has almost the same physiological characteristic with Xiangyou 15 variety. To find out the difference between high oleic acid rapeseed material and Xiangyou 15 seedling, a comparative proteomic approach based on 2-DE and mass spectrometry was adopted. A total of 277 protein spots showed a significant change in intensity by more than 2.0-fold from M15 compared with Xiangyou 15 variety. Among them, 48 spots that changed at least 3.0-fold were excised from gels and successfully identified by MALDI-TOF/TOF MS. The identified proteins involved in metabolism of carbohydrate and energy （75%）, stress and defense （8.3%）, photosynthesis （6.3%）, protein metabolism （2.1%） and other functions （8.3%）. Then real-time quantitative PCR （qPCR） analysis was used to verify the expression levels of differentially expressed proteins, but the results did well agree with the proteomic results. In this work, most of the proteins involved in metabolism of carbohydrate and energy have higher expression in M15, which may reveal M15 has higher metabolism ability. These results provided much information to understand the differences between high oleic acid rapeseed material and Xiangyou 15 variety, which will be useful to screen high oleic rapeseed materials in seedling period.

关键词： proteomic analysis high oleic acid 2-D electrophoresis real-time quantitative PCR

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proteomic analysis of differentially expressed proteins involving in liver metastasis of human colorectal carcinoma

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Hepatobiliary & Pancreatic Diseases International 2010年第2期9卷 149-153页

作者： Li, Shi-Yong An, Ping Cai, Hui-Yun Bai, Xue Zhang, Ying-Nan Yu, Bo Zuo, Fu-Yi Chen, Gang Gen Hosp Beijing Mil Command Dept Gen Surg Beijing 100700 Peoples R China

BACKGROUND: Early diagnosis of liver metastasis of colorectal carcinoma is very important for the appropriate treatment of such patients. However, there has been no effective approach available for clinical application. The present study aimed to investigate the differential expression of proteins in patients with liver metastasis of colorectal carcinomas using proteomic analysis and evaluate its potentiality in clinical diagnosis. METHODS: Fluorescence two-dimensional differential in-gel electrophoresis (2-D DIGE) was used to analyze and compare the protein expression between normal mucosa, the primary focus, and liver metastases. proteomic analysis was made to identify the differentially expressed proteins. Immunohistological staining was used to confirm the expression of differentially expressed proteins in colorectal carcinomas and areas of liver metastasis. RESULTS: A 1.5-fold difference was found with 46 differentially expressed proteins. In 20 differentially expressed proteins, 3 were down-regulated and 17 up-regulated in liver metastases. proteomic analysis showed that the S-adenosylmethionine transgelin variant was down-regulated in liver metastasis tissues. Zinc finger protein 64 homolog (Zfp64), guanine nucleotide exchange factor 4 (GEF4), human arginase, glutathione S-transferases (GSTs) A3, and tumor necrosis factor a (TNF-alpha)-induced protein 9 were up-regulated in liver metastasis tissues. Immunohistochemical staining confirmed that human arginase expression was higher in liver metastases than in the primary focus. CONCLUSIONS: There was a significant difference in protein expression between the primary focus of colorectal carcinoma and liver metastases. The differentially regulated proteins were closely related to liver metastasis of colorectal carcinoma. Elevated human arginase may be an important molecular marker for liver metastasis from colorectal carcinoma. (Hepatobiliary Pancreat Dis Jut 2010; 9: 149-153)

关键词： colorectal carcinoma liver metastasis proteomic analysis human arginase

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proteomic analysis of anti-angiogenic effects by conbercept in the mice with oxygen induced retinopathy

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International Journal of Ophthalmology(English edition) 2020年第12期13卷 1844-1853页

作者： Ji Jin Lei Chen Gao-Qin Liu Pei-Rong Lu Department of Ophthalmology the First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina Department of Ophthalmology the Second Affiliated Hospital of Soochow UniversitySuzhou 215004Jiangsu ProvinceChina Department of Ophthalmology Children's Hospital of Soochow UniversitySuzhou 215025Jiangsu ProvinceChina

AIM:To analyze the retinal proteomes with and without conbercept treatments in mice with oxygen-induced retinopathy(OIR)and identify proteins involved in the molecular mechanisms mediated by conbercept.METHODS:OIR was induced in fifty-six C57 BL/6 J mouse pups and randomly divided into four groups.Group 1:Normal17(n=7),mice without OIR and treated with normal air.Group 2:OIR12/EXP1(n=14),mice received 75%oxygen from postnatal day(P)7 to 12.Group 3:OIR17/Control(n=14),mice received 75%oxygen from P7 to P12 and then normal air to P17.Group 4:Lang17/EXP2(n=21),mice received 75%oxygen from P7 to P12 with intravitreal injection of 1μL conbercept at the concentration of 10 mg/m L at P12,and then normal air from P12 to P17.Liquid ChromatographyMass Spectrometry(LC-MS)/MS data were reviewed to find proteins that were up-regulated after the conbercept treatment.Gene ontology(GO)analysis was performed of conbercept-mediated changes in proteins involved in single-organism processes,biological regulation,cellular processes,immune responses,metabolic processes,locomotion and multiple-organism processes.RESULTS:Conbercept induced a reversal of hypoxiainducible factor 1 signaling pathway as revealed by the Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis and also induced down-regulation of proteins involved in blood coagulation and fibrin clot formation as demonstrated by the Database for Annotation,Visualization and Integrated Discovery(DAVID)and the stimulation of interferon genes studies.These appear to be risk factors of retinal fibrosis.Additional conbercept-specific fibrosis risk factors were also identified and may serve as therapeutic targets for fibrosis.CONCLUSION:Our studies reveal that many novel proteins are differentially regulated by conbercept.The new insights may warrant a valuable resource for conbercept treatment.

关键词： anti-vascular endothelial growth factor conbercept retinopathy fibrosis proteomic analysis

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iTRAQ-based quantitative proteomic analysis on differentially expressed proteins of rat mandibular condylar cartilage induced by reducing dietary loading

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Frontiers of Medicine 2017年第1期11卷 97-109页

作者： Liting Jiang Yinyin Xie Li Wei Qi Zhou Ning Li Xinquan Jiang Yiming Gao Department of Stomatology Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Department of Prosthodontics Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai Key Laboratory of Stomatology Shanghai 200011 China State Key Laboratory of Medical Genomics Shanghai Institute of Hematology Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Shanghai Institute of Traumatology and Orthopedics Shanghai 200025 China

As muscle activity during growth is considerably important for mandible quality and morphology, reducing dietary loading directly influences the development and metabolic activity of mandibular condylar cartilage （MCC）. However, an overall investigation of changes in the protein composition of MCC has not been fully described in literature. To study the protein expression and putative signaling in vivo, we evaluated the structural changes of MCC and differentially expressed proteins induced by reducing functional loading in rat MCC at developmental stages. Isobaric tag for relative and absolute quantitation-based 2D nano-high performance liquid chromatography （HPLC） and matrix-assisted laser desorption/ionization time-of-flight/ time-of-flight （MALDI-TOF/TOF） technologies were used. Global protein profiling, KEGG and PANTHER pathways, and functional categories were analyzed. Consequently, histological and tartrate-resistant acid phosphatase staining indicated the altered histological structure of condylar cartilage and increased bone remodeling activity in hard-diet group. A total of 805 differentially expressed proteins were then identified. GO analysis revealed a significant number of proteins involved in the metabolic process, cellular process, biological regulation, localization, developmental process, and response to stimulus. KEGG pathway analysis also suggested that these proteins participated in various signaling pathways, including calcium signaling pathway, gap junction, ErbB signaling pathway, and mitogen-activated protein kinase signaling pathway. Collagen types I and II were further validated by immunohistochemical staining and Western blot analysis. Taken together, the present study provides an insight into the molecular mechanism of regulating condylar growth and remodeling induced by reducing dietary loading at the protein level.

关键词： condylar cartilage mechanical loading proteomic analysis iTRAQ bioinformatics analysis

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proteomic analysis of exosomes isolated from saliva and salivary glands of tick Haemaphysalis longicornis

Proteomic analysis of exosomes isolated from saliva and sali...

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中国畜牧兽医学会兽医寄生虫学分会第一届青年科学家学术论...

作者： Mohsin Nawaz Muhammad Irfan Malik Houshuang Zhang Ibrahim A.Hassan Jie Cao Yongzhi Zhou Mudassar Hameed Zulfiqar Hussain Kuthu Jinlin Zhou key Laboratory of Animal Parasitology of Ministry of Agriculture Shanghai Veterinary Research InstituteChinese Academy of Agricultural Sciences

Background:Exosomes,the nano-sized membrane vesicles are considered as vehicles of cellular communication.Parasites usually release exosomes in their exeretory-secretory products to modulate host environment.However,a little is known about the secretion of exosomes by ticks.In this study,we showed for the very first time that the tick,Haemaphysalis longicornis,secretes micro vesicles,resembling to exosomes.Methods:Exosomes were purified from saliva and salivary glands of H.longicornis ticks,through serial steps of ultracentrifugation.Presence of exosomes was confirmed by electron microscopy.Liquid chromatography(LC-MS/MS) was used to characterize the proteins within tick-derived exosomes.In addition,GO and KEGG database were mapped to analyse the molecular functions,biological processes,cellular components and pathways associated with salivary-exosomal proteins.Results:Electron microscopy analysis of saliva and culture medium containing salivary glands revealed the presence of exosomes like vesicles within a size range of 50-100 nm.Presence of exosomal markers(Hsp70 and elongation factor 1) as well as parasite specific proteins gave us further confirmation of their release by the ticks.A total of 354 proteins in salivary exosomes whereas 287 proteins in culture medium containing salivary glands were identified.Most of the proteins within tick-derived exosomes have been identified as components of parasitic exosomes.Bioinformatics analysis(Gene Ontology) indicated association of certain biological and molecular functions with salivary exosomal proteins which may be helpful in development of ticks.Likewise,KEGG database revealed involvement of salivary exosomal proteins in proton transport,detoxification,ECM-receptor interaction,Ribosome,RNA transport,ABC transporters and oxidative phosphorylation.Conclusion:Results of this study provide evidence that exosomes are being secreted by the ticks and suggests that tick derived exosomes could play important roles in host parasite relationship.Moreover,exosomes could be u

关键词： Haemaphysalis longicornis exosomes salivary glands proteomic analysis

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proteomic analysis of hippocampal phosphoprotein reveals up-regulation of phosphorylated 14-3-3 theta in F344 rats exposed to 1-bromopropane

Proteomic analysis of hippocampal phosphoprotein reveals up-...

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中国毒理学会第六届全国毒理学大会

作者： HUANG Zhen-lie Gaku ICHIHARA Sahoko ICHIHARA CHANG Jie ZHANG Ling-yi Guangdong Prevention and Treatment Center for Occupational Diseases Nagoya University Graduate School of Medicine Mie University Graduate School of Regional Innovation Studies

OBJECTIVE 1-Bromopropane(1-BP) has been introduced as an alternative toozone-depleting solvents and exhibits neurotoxicity both in experimental animals and human.To obtain further information on the underlying mechanisms of 1-BP-induced neurotoxicity in the central nerve system (CNS),a proteomic analysis was conducted to identify and characterize 1-BP-induced modification of phosphoprotein expression in the hippocampus of rats.METHODS Male F344 rats were exposed to 1-BP at 0,400,or 1000 ppm for 8 h·d for 1 week and 4 weeks by inhalation(n =9 rats of each group).Hippocampal protein extracts(n=3 rats of each group) from control and 1000 ppm in 4-week exposure were resolved by two-dimensional electrophoresis(2DE),Pro-Q Diamond Phosphoprotein Gel Staining and SYPRO gel staining for qualitation of phosphoproteins.Two-dimensional difference in gel electrophoresis(2D-DIGE) was conducted to quantitatively analysis of phosphoprotein(n=6 rats in each group) and matrix-assisted laser-desorption ionization time-of-flight(MALDI-TOF) mass spectrometry (MS) was used for protein identification.Selected protein was further confirmed by Phos-tag SDS-PAGE. RESULTS 2D-DIGE revealed 28 protein spots in 1000 ppm group of 4-week 1-BP exposure with statistically significant difference(P<0.05) compared with the controls,including 21 up-regulated spots and 7 down-regulated spots,which were detected as phosphoproteins by Pro-Q Diamond gel staining combined with SYPRO staining.MALDI-TOF-TOF/MS successfully identified 19 up-regulated spots and 7 down-regulated spots.These identified proteins participate in the biological processes of metabolic, immune,response to stimulus,transport and cellular communication,and involved in pathways including Parkinson’s disease,apoptosis signaling,FGF and EGF receptor signaling pathway,etc.Expression of phosphorylated 14-3-3 theta protein was further confirmed by Phos-tag SDS-PAGE.CONCLUSION These results help us understand the pro-apoptosis mechanism of 1 -BP-induced hippocampal response and establish pote

关键词： 1-bromopropane proteomic analysis phosphoprotein

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