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Nutritional stimulation by in-ovo feeding modulates cellular proliferation and differentiation in the small intestinal epithelium of chicks

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Animal Nutrition 2022年第1期8卷 91-101页

作者： Naama Reicher Tal Melkman-Zehavi Jonathan Dayan Eric AWong Zehava Uni Department of Animal Science The Robert H.SmithFaculty of AgricultureFood and EnvironmentThe Hebrew University of JerusalemRehovot 76100Israel Department of Animal and Poultry Sciences Virginia TechBlacksburgVA 24061USA

Nutritional stimulation of the developing small intestine of chick embryos can be conducted by in-ovo feeding(IOF).We hypothesized that IOF of glutamine and leucine can enhance small intestinal development by promoting proliferation and differentiation of multipotent small intestinal epithelial cells.Broiler embryos(n=128)were subject to IOF of glutamine(IOF-Gln),leucine(IOF-Leu),NaCl(IOF-NaCl)or no injection(control)at embryonic d 17(E 17).Multipotent,progenitor and differentiated cells were located and quantified in the small intestinal epithelium between E 17 and d 7 after hatch(D 7)in all treatment groups by immunofluorescence of SRY-box transcription factor 9(Sox9)and proliferating cell nuclear antigen(PCNA),in-situ hybridization of leucine-rich repeat containing G-protein coupled receptor 5(Lgr5)and peptide transporter 1(PepT1)and histochemical goblet cell staining.The effects of IOF treatments at E 19(48 h post-IOF),in comparison to control embryos,were as follows:total cell counts increased by 40%,33%and 19%,and multipotent cell counts increased by 52%,50%and 38%,in IOF-Gln,IOF-Leu and IOF-NaCl embryos,respectively.Only IOF-Gln embryos exhibited a significance,36%increase in progenitor cell counts.All IOF treatments shifted Lgr5+ stem cell localizations to villus bottoms.The differentiated,PepT1+region of the villi was 1.9 and 1.3-fold longer in IOF-Gln and IOF-Leu embryos,respectively,while goblet cell densities decreased by 20%in IOF-Gln embryos.Postehatch,crypt and villi epithelial cell counts were significantly higher IOF-Gln chicks,compared to control chicks(Pdifferentiation,resulting in enhanced compartmentalization of multipotent and differentiated cell niches and expansions of the absorptive surface area.

关键词： Chick Intestinal development Proliferation differentiation Multipotent cells In-ovo feeding

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miR-103-3p targets Ndel1 to regulate neural stem cell proliferation and differentiation

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Neural Regeneration Research 2022年第2期17卷 401-408页

作者： Wen Li Shan-Shan Wang Bo-Quan Shan Jian-Bing Qin He-Yan Zhao Mei-Ling Tian Hui He Xiang Cheng Xin-Hua Zhang Guo-Hua Jin Department of Human Anatomy Institute of NeurobiologyNantong UniversityNantongJiangsu ProvinceChina Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education NantongJiangsu ProvinceChina Co-Innovation Center of Neuroregeneration Nantong UniversityNantongJiangsu ProvinceChina

The regulation of adult neural stem cells(NSCs) is critical for lifelong neurogenesis. MicroRNAs(miRNAs) are a type of small, endogenous RNAs that regulate gene expression post-transcriptionally and influence signaling networks responsible for several cellular processes. In this study, mi R-103-3 p was transfected into neural stem cells derived from embryonic hippocampal neural stem cells. The results showed that mi R-103-3 p suppressed neural stem cell proliferation and differentiation, and promoted apoptosis. In addition, mi R-103-3 p negatively regulated Nud E neurodevelopment protein 1-like 1(Ndel1) expression by binding to the 3′ untranslated region of Ndel1. Transduction of neural stem cells with a lentiviral vector overexpressing Ndel1 significantly increased cell proliferation and differentiation, decreased neural stem cell apoptosis, and decreased protein expression levels of Wnt3 a, β-catenin, phosphor-GSK-3β, LEF1, c-myc, c-Jun, and cyclin D1, all members of the Wnt/β-catenin signaling pathway. These findings suggest that Ndel1 is a novel mi R-103-3 p target and that mi R-103-3 p acts by suppressing neural stem cell proliferation and promoting apoptosis and differentiation. This study was approved by the Animal Ethics Committee of Nantong University, China(approval No. 20200826-003) on August 26, 2020.

关键词： apoptosis canonical Wnt pathway differentiation MiR-103-3p Ndel1 neural stem cells neurogenesis proliferation

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Poly-L-ornithine blocks the inhibitory effects of fibronectin on oligodendrocyte differentiation and promotes myelin repair

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Neural Regeneration Research 2023年第4期18卷 832-839页

作者： Ya-Jie Xiong Shahid Hussain Soomro Zhong-Hai Huang Pan-Pan Yu Jie Ping Hui Fu Department of Anatomy School of Basic Medical SciencesWuhan UniversityWuhanHubei ProvinceChina Guangdong-Hongkong-Macao Institute of CNS Regeneration Ministry of Education Joint International Research Laboratory of CNS RegenerationJinan UniversityGuangzhouGuangdong ProvinceChina Department of Pharmacology School of Basic Medical SciencesWuhan UniversityWuhanHubei ProvinceChina

The extracellular matrix surrounding oligodendrocytes plays an important role during myelination and remyelination in the brain.In many cases,the microenvironment surrounding demyelination lesions contains inhibitory molecules,which lead to repair failure.Accordingly,blocking the activity of these inhibitory factors in the extracellular matrix should lead to more successful remyelination.In the central nervous system,oligodendrocytes form the myelin sheath.We performed primary cell culture and found that a natural increase in fibronectin promoted the proliferation of oligodendrocyte progenitors during the initial stage of remyelination while inhibiting oligodendrocyte differentiation.Poly-L-ornithine blocked these inhibitory effects without compromising fibronectin’s pro-proliferation function.Experiments showed that poly-L-ornithine activated the Erk1/2 signaling pathway that is necessary in the early stages of differentiation,as well as PI3K signaling pathways that are needed in the mid-late stages.When poly-L-ornithine was tested in a lysolecithin-induced animal model of focal demyelination,it enhanced myelin regeneration and promoted motor function recovery.These findings suggest that poly-L-ornithine has the potential to be a treatment option for clinical myelin sheath injury.

关键词： differentiation Erk1/2 extracellular matrix fibronectin lysolecithin-induced demyelination oligodendrocyte PI3K poly-L-ornithine proliferation remyelination

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The developmental changes in intestinal epithelial cell proliferation,differentiation,and shedding in weaning piglets

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Animal Nutrition 2022年第2期9卷 214-222页

作者： Min Wang Lixia Wang Xian Tan Lei Wang Xia Xiong Yancan Wang Qiye Wang Huansheng Yang Yulong Yin Hunan International Joint Laboratory of Animal Intestinal Ecology and Health Laboratory of Animal Nutrition and Human HealthCollege of Life SciencesHunan Normal UniversityChangsha 410081HunanChina Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process Hunan Provincial Engineering Research Center for Healthy Livestock and Poultry ProductionKey Laboratory of Agro-ecological Processes in Subtropical RegionInstitute of Subtropical AgricultureChinese Academy of SciencesChangsha 410125HunanChina

Intestinal epithelial homeostasis plays an important role in intestinal morphology and function.However,the developmental changes in intestinal epithelial cell turnover in piglets during early weaning are unknown so far.Thus,the aim of this work was to detect changes in piglet gut development from weaning to post-weaning d 14.Accordingly,40 piglets were used in the present study,and 8 piglets were randomly selected for sampling at d 0,1,3,7 and 14 post-weaning,respectively.The results showed that weaning stress significantly affected small intestinal morphological architecture,and this impact was the worst on d 3,and then returned to normal on d 14.Furthermore,the number of the marker of proliferation Ki-67(Ki67)positive cells was decreased on d 1 and 3,and then recovered on d 14(Pdifferentiation(Pdifferentiation and shedding,and the mTOR signaling pathway was involved in this process.Our findings provide a cellular mechanism for intestinal developmental changes during weaning periods.This may provide nutritionists with better insight into designing efficient in-feed alternatives for preventing the unfavorable gut development in weaning piglets.

关键词： Intestinal epithelial cell Proliferation differentiation Shedding Weaning piglet

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Deciphering the chromatin spatial organization landscapes during BMMSC differentiation

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Journal of Genetics and Genomics 2023年第4期50卷 264-275页

作者： Zhaowei Teng Yun Zhu Da Lin Qinggang Hao Qiaoning Yue Xiaochao Yu Shuo Sun Lihong Jiang Sheng Lu Department of Orthopedics The First People's Hospital of Yunnan ProvinceAffiliated Hospital of Kunming University of Science and TechnologyKunmingYunnan 650032China Key Laboratory of Yunnan Provincial Innovative Application of Traditional Chinese Medicine The First People's Hospital of Yunnan ProvinceKunmingYunnan 650032China Clinical Medical Research Center The First People's Hospital of Yunnan ProvinceKunmingYunnan 650032China The Sixth Affiliated Hospital of Kunming Medical University YuxiYunnan 653100China State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhanHubei 430070China Center for Life Sciences School of Life SciencesYunnan UniversityKunmingYunnan 650504China

The differentiation imbalance in bone marrow mesenchymal stem cells(BMMSCs)is critical for the development of bone density diseases as the population ages.BMMSCs are precursor cells for osteoblasts and adipocytes;however,the chromatin organization landscapes during BMMSC differentiation remain elusive.In this study,we systematically delineate the four-dimensional genome and dynamic epigenetic atlas of BMMSCs by RNA sequencing,assay for transposase-accessible chromatin sequencing,and highthroughput chromosome conformation capture.The structure analyses reveal 17.5%common and28.5%-30%specific loops among BMMSCs,osteoblasts,and adipocytes.The subsequent correlation of genome-wide association studies and expression quantitative trait locus(e QTL)data with multi-omics analysis reveal 274 genes and 3634 single nucleotide polymorphisms(SNPs)associated with bone degeneration and osteoporosis(OP).We hypothesize that SNP mutations affect transcription factor(TF)binding sites,thereby affecting changes in gene expression.Furthermore,26 motifs,260 TFs,and 291SNPs are identified to affect the e QTL.Among these genes,DAAM2,TIMP2,and TMEM241 are found to be essential for diseases such as bone degeneration and OP and may serve as potential drug targets.

关键词： Bone marrow mesenchymal stem cells differentiation Hi-C ATAC-Seq Single nucleotide polymorphisms Genome-wide association studies

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Overexpression of GATA binding protein 4 and myocyte enhancer factor 2C induces differentiation of mesenchymal stem cells into cardiac-like cells

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World Journal of Stem Cells 2022年第9期14卷 700-713页

作者： Syeda Saima Razzaq Irfan Khan Nadia Naeem Asmat Salim Sumreen Begum Kanwal Haneef University of Karachi Karachi 75270Pakistan Panjwani Center for Molecular Medicine and Drug Research International Center for Chemical and Biological SciencesUniversity of KarachiKarachi 75270Pakistan Dow Research Institute of Biotechnology&Biomedical Sciences(DRIBBS) Dow University of Health Sciences(DUHS)Ojha CampusKarachi 75200Pakistan Stem Cells Research Laboratory(SCRL) Sindh Institute of Urology and Transplantation(SIUT)Karachi 74200Pakistan

BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as a new area to support heart regeneration.The transcription factors GATA binding protein 4(GATA-4)and myocyte enhancer factor 2C(MEF2C)are considered prominent factors in the development of the cardiovascular system.AIM To explore the potential of GATA-4 and MEF2C for the cardiac differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs).METHODS hUC-MSCs were characterized morphologically and immunologically by the presence of specific markers of MSCs via immunocytochemistry and flow cytometry,and by their potential to differentiate into osteocytes and adipocytes.hUC-MSCs were transfected with GATA-4,MEF2C,and their combination to direct the differentiation.Cardiac differentiation was confirmed by semiquant itative real-time polymerase chain reaction and immunocytochemistry.RESULTS hUC-MSCs expressed specific cell surface markers CD105,CD90,CD44,and vimentin but lack the expression of CD45.The transcription factors GATA-4 and MEF2C,and their combination induced differentiation in hUC-MSCs with significant expression of cardiac genes i.e.,GATA-4,MEF2C,NK2 homeobox 5(NKX2.5),MHC,and connexin-43,and cardiac proteins GATA-4,NKX2.5,cardiac troponin T,and connexin-43.CONCLUSION Transfection with GATA-4,MEF2C,and their combination effectively induces cardiac differentiation in hUC-MSCs.These genetically modified MSCs could be a promising treatment option for heart diseases in the future.

关键词： Heart disease GATA binding protein 4 Myocyte enhancer factor 2C Transcription factors differentiation Human umbilical cord-mesenchymal stem cells

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E26 transformation-specific 1 is implicated in the inhibition of osteogenic differentiation induced by chronic high glucose by directly regulating Runx2 expression

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The Journal of Biomedical Research 2022年第1期36卷 49-57页

作者： Wenqian Xia Xiao Han Lin Wang Key Laboratory of Human Functional Genomics of Jiangsu Province Nanjing Medical UniversityNanjingJiangsu 211166China Jiangsu Key Laboratory of Oral Diseases Nanjing Medical UniversityNanjingJiangsu 210009China The Affiliated Nantong Stomatological Hospital of Nantong University NantongJiangsu 226001China

Chronic high glucose(HG) plays a crucial role in the pathogenesis of diabetes-induced osteoporosis by inhibiting the differentiation and proliferation of osteoblasts. This study aims to examine the role of E26 transformation-specific 1(ETS1) in the inhibition of osteoblast differentiation and proliferation caused by chronic HG, as well as the underlying mechanism. Chronic HG treatment downregulated ETS1 expression and inhibited differentiation and proliferation of MC3 T3-E1 cells. Downregulation of ETS1 expression inhibited the differentiation and proliferation of MC3 T3-E1 cells under normal glucose conditions, and ETS1 overexpression attenuated the damage to cells exposed to chronic HG. In addition, ETS1 overexpression reversed the decrease in runt-related transcription factor 2(Runx2) expression in MC3 T3-E1 cells treated with chronic HG. Using chromatin immunoprecipitation(ChIP) and luciferase reporter assays, we confirmed that ETS1 directly bound to and increased the activity of the Runx2 promoter. In summary, our study suggested that ETS1 was involved in the inhibitory effect of chronic HG on osteogenic differentiation and proliferation and may be a potential therapeutic target for diabetes-induced osteoporosis.

关键词： ETS1 Runx2 chronic high glucose osteoblast differentiation proliferation

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Serum biomarkers for the differentiation of autoimmune pancreatitis from pancreatic ductal adenocarcinoma

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World Journal of Gastrointestinal Oncology 2023年第2期15卷 268-275页

作者： Octavio Caba Carmelo Diéguez-Castillo Joaquina Martínez-Galán Irene González-Cebrián Cristina Jiménez-Luna Institute of Biopathology and Regenerative Medicine(IBIMER) University of GranadaGranada 18016Spain Biosanitary Institute of Granada(ibs.GRANADA) 18014 GranadaSpain Department of Gastroenterology San Cecilio University HospitalGranada 18016Spain Department of Medical Oncology Virgen de las Nieves University HospitalGranada 18014Spain.Biosanitary Institute of Granada(ibs.GRANADA)18014 GranadaSpain Department of Digestive Cancer San Cecilio University HospitalGranada 18016Spain

Autoimmune pancreatitis(AIP),a chronic inflammation caused by the immune system attacking the pancreas,usually presents imaging and clinical features that overlap with those of pancreatic ductal adenocarcinoma(PDAC).Serum biomarkers,substances that quantitatively change in sera during disease development,are a promising non-invasive tool with high utility for differentiating between these diseases.In this way,the presence of AIP is currently suspected when serum concentrations of immunoglobulin G4(IgG4)antibody are elevated.However,this approach has some drawbacks.Notably,IgG4 antibody concentrations are also elevated in sera from some patients with PDAC.This review focuses on the most recent and relevant serum biomarkers proposed to differentiate between AIP and PDAC,evaluating the usefulness of immunoglobulins,autoantibodies,chemokines,and cytokines.The proposed serum biomarkers have proven useful,although most studies had a small sample size,did not examine their presence in patients with PDAC,or did not test them in humans.In addition,current evidence suggests that a single serum biomarker is unlikely to accurately differentiate these diseases and that a set of biomarkers will be needed to achieve adequate specificity and sensitivity,either alone or in combination with clinical data and/or radiological images.

关键词： Autoimmune pancreatitis Pancreatic ductal adenocarcinoma Serum Biomarkers differentiation

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MicroRNA-146a Promotes Embryonic Stem Cell differentiation towards Vascular Smooth Muscle Cells through Regulation of Kruppel-like Factor 4

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Current Medical Science 2023年第2期43卷 223-231页

作者： Qing ZHANG Rong-rong PAN Yu-tao WU Yu-miao WEI Department of Cardiology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China Hubei Key Laboratory of Biological Targeted Therapy Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China Department of Cardiology Affiliated Cixi HospitalWenzhou Medical UniversityNingbo 315300China Department of Cardiology First Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhou 310003China

Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.

关键词： microRNA-146a embryonic stem cells differentiation vascular smooth muscle cells Kruppel-like factor 4

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MiR-743a-5p regulates differentiation of myoblast by targeting Mob1b in skeletal muscle development and regeneration

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Genes & Diseases 2022年第4期9卷 1038-1048页

作者： YongSheng Zhang YiLong Yao ZiShuai Wang Dan Lu YuanYuan Zhang Adeyinka Abiola Adetula SiYuan Liu Min Zhu YaLan Yang XinHao Fan MuYa Chen YiJie Tang Yun Chen YuWen Liu GuoQiang Yi ZhongLin Tang Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenGuangdong 518124PR China Genome Analysis Laboratory of the Ministry of Agriculture Agricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenGuangdong 518124PR China Institute of Animal Science Chinese Academy of Agricultural SciencesBeijing 100193PR China

The microRNAs (miRNAs) play an important role in regulating myogenesis by targeting mRNA. However, the understanding of miRNAs in skeletal muscle development and diseases is unclear. In this study, we firstly performed the transcriptome profiling in differentiating C2C12 myoblast cells. Totally, we identified 187 miRNAs and 4260 mRNAs significantly differentially expressed that were involved in myoblast differentiation. We carried out validation of microarray data based on 5 mRNAs and 5 miRNAs differentially expressed and got a consistent result. Then we constructed and validated the significantly up- and down-regulated mRNA-miRNA interaction networks. Four interaction pairs (miR-145a-5p-Fscn1, miR-200c-5p-Tmigd1, miR-27a-5p-Sln and miR-743a-5p-Mob1b) with targeted relationships in differentiated myoblast cells were demonstrated. They are all closely related to myoblast development. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated cell cycle signals important for exploring skeletal muscle development and disease. Functionally, we discovered that miR-743a targeting gene Mps One Binder Kinase Activator-Like 1B (Mob1b) gene in differentiated C2C12. The up-regulated miR-743a can promote the differentiation of C2C12 myoblast. While the down-regulated Mob1b plays a negative role in differentiation. In addition, the expression profile of miR-743a and Mob1b are consistent with skeletal muscle recovery after Cardiotoxin (CTX) injury. Our study revealed that miR-743a-5p regulates myoblast differentiation by targeting Mob1b involved in skeletal muscle development and regeneration. Our findings made a further exploration for mechanisms in myogenesis and might provide potential possible miRNA-based target therapies for skeletal muscle regeneration and disease in the near future.

关键词： differentiation miR 743a-5p Mob 1b Myoblast Skeletal muscle .Transcriptome

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