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Gymnema montanum improves endothelial function via inhibition of endoplasmic reticulum stress by activating Nrf2 signaling

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Asian Pacific Journal of Tropical Biomedicine 2020年第8期10卷 379-386页

作者： Dornadula Sireesh Natarajan Suganya Suvro Chatterjee Kunka Mohanram Ramkumar SRM Research Institute SRM Institute of Science and TechnologyKattankulathur-603203Tamil NaduIndia Department of Biotechnology School of BioengineeringSRM Institute of Science and TechnologyKattankulathur-603203Tamil NaduIndia Vascular Biology Lab AU-KBC Research CentreAnna UniversityChromepetChennai-600044Tamil NaduIndia

Objective:To investigate the effects of Gymnema montanum leaf extract against endoplasmic reticulum(ER)stress-induced toxicity in endothelial cells.Methods:The immortalized endothelial hybrid cell,EA.hy926 was treated with different concentrations of Gymnema montanum leaf extract(0–100μg/mL)and the ER stress inducer,tunicamycin.The cytotoxicity was assessed by MTT as well as lactate dehydrogenase and malondialdehyde levels were determined.The levels of ER stress markers,GRP78 and CHOP were analysed by Western blot assay.The Gymnema montanum leaf extract-mediated activation of nuclear factor erythroid 2-related factor 2(Nrf2)was assessed by cell-based luciferase enzyme fragment complementation assay and antioxidant responsive element driven luciferase reporter assay.The levels of phosphoproteins of the MAPK pathway were analyzed using the Bioplex system.Results:A dose-dependent cytoprotective effect of Gymnema montanum leaf extract was observed in tunicamycin-induced toxicity.Gymnema montanum leaf extract significantly reduced lactate dehydrogenase activity and malondialdehyde levels in ER stress-induced endothelial cells.It also suppressed ER stress markers dose dependently and inhibited the phosphorylation of JNK,ERK,MEK and p38 MAPK in tunicamycin-induced endothelial cells.Moreover,Gymnema montanum leaf extract increased the expression of Nrf2 and its downstream targets in endothelial cells.Conclusions:Gymnema montanum leaf extract attenuates ER stress by increasing the expression of Nrf2 and its downstream genes.

关键词： ER stress Gymnema montanum Nrf2 endothelial cells

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The research of HSPB1 in apoptosis,autophagy and cardiogenic shock

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实用休克杂志（中英文） 2023年第6期7卷 380-384页

作者： Gu Xiaoxiao Jiang Weibin Chen Fang Liu Xiehong Hunan provincial key laboratory of emergency and critical care metabonomics Hunan provincial peoples hospital(The first affiliated hospital of Hunan normal university) The first affiliated hospital of Hunan normal university ChangshaChina

Cardiogenic shock(CS)is a common cause of mortality and treatment remains challenging despite advances in therapeutic options.CS is caused by severe impairment of myocardial performance that results in decreased cardiac output,hypoperfusion of the end organ,and hypoxia.Clinically this presents as hypotension refractory to volume resuscitation with features of end-organ hypoperfusion requiring pharmacological or mechanical intervention.Acute myocardial infarction(AMI)accounts for 81%of patients with CS.Heat shock protein family B member 1(HSPB1)is a multifunctional protein induced by various stress factors and has a protective effect on cells.A large number of studies have demonstrated that HSPB1 plays an important role in regulating apoptosis.Recently,some studies have suggested that HSPB1 also participates in the autophagic process.HSPB1 are expressed in many cells of the cardiovascular system such as endothelial cells,cardiac muscle cells,monocytes,and platelets.They are up-regulated in response to inflammation,oxidative stress,or ischemia and protect cells against extracellular stress factors.Here,we explore the involvement of HSPB1 in apoptosis,autophagy,and CS.We speculate that HSPB1 may exert its anti-myocardial injury role via the regulation of apoptosis and autophagy;this may provide the basis for the development of new approaches for the prevention and treatment of CS.

关键词： Cardiogenic shock Acute myocardial infarction HSPB1 endothelial cells Cardiac muscle cells Apoptosis Autophagy

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Vascularization mediated by mesenchymalstem cells from bone marrow and adiposetissue: a comparison

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Cell Regeneration 2015年第1期4卷 106-115页

作者： Karoline Pill Sandra Hofmann Heinz Redl Wolfgang Holnthoner Ludwig Boltzmann Institute for Experimental and Clinical Traumatology Donaueschingenstrasse 131200ViennaAustria Austrian Cluster for Tissue Regeneration ViennaAustria Department of Biomedical Engineering Eindhoven University of Technology5600 MB EindhovenThe Netherlands Institute for Complex Molecular Systems Eindhoven University of TechnologyPO Box 5135600 MB EindhovenThe Netherlands Institute for Biomechanics Swiss Federal Institute of Technology ZürichVladimir-Prelog-Weg 38093 ZürichSwitzerland

Tissue-engineered constructs are promising to overcome shortage of organ donors and to reconstruct at least partsof injured or diseased tissues or organs. However, oxygen and nutrient supply are limiting factors in many tissues,especially after implantation into the host. Therefore, the development of a vascular system prior to implantationappears crucial. To develop a functional vascular system, different cell types that interact with each other need tobe co-cultured to simulate a physiological environment in vitro. This review provides an overview and a comparison ofthe current knowledge of co-cultures of human endothelial cells (ECs) with human adipose tissue-derived stem/stromal cells (ASCs) or bone marrow-mesenchymal stem cells (BMSCs) in three dimensional (3D) hydrogel matrices.Mesenchymal stem cells (MSCs), BMSCs or ASCs, have been shown to enhance vascular tube formation of ECs and toprovide a stabilizing function in addition to growth factor delivery and permeability control for ECs. Althoughphenotypically similar, MSCs from different tissues promote tubulogenesis through distinct mechanisms. In this report,we describe differences and similarities regarding molecular interactions in order to investigate which of these two celltypes displays more favorable characteristics to be used in clinical applications. Our comparative study shows that ASCsas well as BMSCs are both promising cell types to induce vascularization with ECs in vitro and consequently arepromising candidates to support in vivo vascularization.

关键词： Vascularization endothelial cells Mesenchymal stem cells Co-culture Molecular mechanisms

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Characterization of Adherent Nonhematopoietic cells Derived from Human Umbilical Cord Blood

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Tsinghua Science and Technology 2003年第4期8卷 412-416页

作者：安小惠蔡国平 Department of Biological Sciences and Technology Tsinghua University Beijing 100084 China

To confirm and characterize the adherent fibroblast like progenitors in human umbilical cord blood, we isolated mononuclear cells from human umbilical cord blood by Ficoll Hypaque. Two main morphologically different kinds of cells were formed by culturing the cells in collagen coated 24 well plastic dishes and flasks. One type was the adherent fibroblast like cells, while the other was loosely adherent clonally expanded round cells. Our experiments demonstrate that the adherent fibroblast like cells possess multilineage potential, including the ability to differentiate into endothelial like cells and to express the mesenchymal cell marker.

关键词： human umbilical cord blood mesenchymal cells endothelial cells

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Construction of eukaryotic expression vector of humanarresten gene and its secreted expression in HEK 293 cells

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Frontiers of Medicine 2009年第3期3卷 297-302页

作者： Wei LI Zifang SONG Qichang ZHENG Jun XIONG Dan SHANG Siming GUAN Xiaogang SHU Department of Gerontology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China Department of General Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

The eukaryotic expression vector of human arresten gene was constructed and its secretive expression in human embryonic kidney(HEK 293)cells was detected.Human arresten gene was ampliﬁed from recombinant plasmid pGEM-Arr by polymerase chain reaction(PCR),and then digested with restriction endonucleases BamH I and EcoR I.The target fragment was inserted into the BamH I and EcoR I restriction sites of eukaryotic expression vector pSecTag2 to construct pST-AT.Restric-tion analysis and DNA sequencing indicated that the arresten gene was successfully inserted into pSecTag2.The recombinant plasmid was subsequently transfected into HEK 293 cells with LipofectAMINETM2000 Reagent,and the expression of the target gene was detected.Reverse transcription PCR(RT-PCR)revealed that the mRNA of the target gene was transcribed in the transfected HEK 293 cells.Western blot analysis veriﬁed that the recombinant protein in supernatants was correct.The supernatants of transfected cells were prepared,and 3-(4,5)-dimethylthiazol(-2-y1)-3,5-di-phenyltetrazoliumbromide(MTT)assay was carried out to assess their effects on the proliferation of human umbilical vein endothelial cells,which showed that the recombinant protein could signiﬁcantly suppress the proliferation of human umbilical vein endothelial cells in vitro.These results provided a solid foundation to explore the usage of arresten in tumor anti-angiogenic gene therapy.

关键词： angiogenesis inhibitor arresten eukaryotic expression HEK 293 cells endothelial cells

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Angiogenesis in acute myeloid leukemia

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Journal of Cancer Metastasis and Treatment 2020年第1期6卷 690-706页

作者： Ugo Testa Germana Castelli Elvira Pelosi Department of Oncology Istituto Superiore di SanitàRome 00161Italy

Angiogenesis is a word that refers to new blood vessel formation,and this process is of fundamental importance for physiological development and tissue homeostasis,as well as the genesis of several diseases,including tumors.Thus,studies carried out in the last years have shown that angiogenesis is essential for the growth of many solid tumors.Angiogenesis is also important for the growth of many hematological malignancies,including acute myeloid leukemia(AML).endothelial cells are essential constituents of the bone marrow vascular niches,structures essential for the survival and maintenance of normal hematopoietic stem/progenitor cells.Bone marrow endothelial cells play an essential role in leukemia development and there is growing evidence that a targeting of both leukemic and endothelial cells of the leukemic vascular niche may improve the efficacy of antileukemic therapies.Bone marrow angiogenesis is frequently increased in AML,is morphologically evidenced as increased microvascular density,and is typically associated with some AML subtypes.The molecular mechanisms underlying the increased angiogenesis in some AML subtypes have been defined.In conclusion,a better understanding of angiogenesis as well as the fundamental interactions between bone marrow endothelial cells and leukemic stem cells may contribute to improve antileukemia treatments.

关键词： Acute myeloid leukemia angiogenesis microvascular density vascular niche endothelial cells endothelial growth factors

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Mechanism of vascular endothelial growth factor on the prevention of restenosis after angioplasty

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Frontiers of Medicine 2009年第2期3卷 177-180页

作者： Qigong LIU Honglian ZHOU Yan ZENG Shan YE Jiani LIU Zaiying LU Department of Internal Cardiology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

To evaluate the mechanism of vascular endothelial growth factor(VEGF)on the prevention of restenosis after angioplasty,the recombinant adenovirus vector containing hVEGF165 cDNA was constructed and transfected into vascular smooth muscle cells(VSMC)in vitro.The conditioned medium containing VEGF was collected 72 h after the infection.Then,the VSMC and human umbilical vein endothelial cells(HUVEC)were divided into control group,H2O2-treated group and H2O2+VEGF-treated group to observe the proliferation and apoptosis by water soluble tetrazolium(WST-1)method,in situ nick end labeling(TUNEL)andﬂow cytometry(FCM).Compared with the control and H2O2+VEGF-treated groups,the absorbance(A)value of HUVEC was decreased,and apoptosis of HUVEC was signiﬁcantly increased in H2O2-treated group.The changes of A value and apoptosis of VSMC were contrary to those of HUVEC.H2O2 could stimulate the proliferation of VSMC and induce the apoptosis of HUVEC,inhibit the proliferation of HUVEC and the apoptosis of VSMC and induce restenosis.VEGF could inhibit the effect of H2O2 on HUVEC and VSMC and prevent restenosis.These results offered further theoretical evidence for VEGF on the prevention of restenosis after angioplasty.

关键词： vascular endothelial growth factors resteno-sis reactive oxygen species endothelial cells vascular smooth muscle cell

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Study of Zonula occluden-1

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Neural Regeneration Research 2006年第6期1卷 553-556页

作者： Yujing Huang Jiachun Feng Department of Neurology First Hospital Jilin University Changchun 130021 Jilin Province China

OBJECTIVE： To investigate the structure, location, function and regulation of Zonula occluden-1 （ZO-1） as well as the correlation of ZO-1 with diseases for helping diagnosis, treatment and determining prognosis of correlative diseases. DATA SOURCES ： Using the terms ＂Zonula Occluden＂, we searched the Medline database to identify ZO-1 related articles published between May 1963 and December 2006 in English. STUDY SELECTION ： The articles were selected firstly, and those involved the structure, location, function and regulation of ZO-1 as well as the correlation of ZO-1 with diseases were chosen. The full texts of the left articles were searched. Inclusive cnteda： Studies associated with the structure, location, function and regulation of ZO-1 as well as those associated with the correlation of ZO-1 with diseases. Exclusive cnteria： Studies with repetitive or uncorrelated contents. DATA EXTRACTION ： Among 1 462 collected articles of ZO-1, 1 424 were excluded. Of the 38 articles, which met inclusive criteria, 11, 2, 10, 11 and 4 were associated with the structure, location, function and regulation of ZO-1 as well as the correlation of ZO-1 with diseases, respectively. DATA SYNTHESIS： ZO-1 is a member of membrane associated guarylate kinasellike proteins （MAGUKS）, can not form homooligomers and exists as ZO-1/ZO-2 and ZO-1/ZO-3 heterodimers. It is expressed in endothelial cells and epithelial cells and also in other types （except for tight junction） of cells. It exists not only in tight junction but also in adherens junction. ＂ Messenger＂ which contacts with various junctional complexes, interacts with tight junction, adherens junction and gap junction to maintain the structure and function of junctional complex. The structure and function of ZO-1 are regulated by many factors, such as Ca^2＋, phosphorylation, protein kinase C, G protein and so on. ZO-1 plays an important role in the treatment of ischemia/hypoxia, inflammation and tumors. CONCLUSION： ZO-1, which is only used as a tight junction protein initi

关键词： adherens junction structure and function gap junction endothelial cells epithelial cells

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Progress Update in Space Cell Mechano-biological Coupling

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空间科学学报 2020年第5期40卷 935-936页

作者： LONG Mian SUN Shujin LI Ning LÜDongyuan GAO Yuxin National Microgravity Laboratory Institute of MechanicsChinese Academy of SciencesBeijing 100190

Recent progresses in 2018–2019 from space experiments onboard SJ-10 recoverable satellite and on parabolic flight were summarized,mainly focusing on cell mechano-biological coupling under microgravity.In the meantime,technical pre-research and experimental system design for the biomechanics research platform on China Space Station was carried out and updated.

关键词： Microgravity SJ-10 satellite Parabolic flight endothelial cells Mesenchymal stem cells Hepatocytes Mechanotransduction China Space Station

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Relationship of endothelial nitric oxide synthase gene polymorphism with blood pressure,lipid profile and blood glucose level

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Frontiers of Medicine 2008年第2期2卷 178-181页

作者： Zhongzhi TANG Junyao LI Jianhong YANG Department of Critical Care Medicine Wuhan General HospitalGuangzhou Command of PLAWuhan 430070China

To study the relationship of the polymorphism of endothelial nitric oxide synthase(eNOS)gene and blood pressure,lipid profiles and blood glucose level.By using PCR-RFLP,the eNOS Glu298Asp gene polymorphism was detected in 184 patients with essential hypertension and 196 matched healthy individuals with normal blood pressure.Taking into account eNOS Glu298Asp poly-morphisms,the relationship of blood pressure with trigly-cerides(TG),total cholesterol(TC),high density lipoprotein(HDL),low density lipoprotein(LDL)and blood glucose level was analyzed.The distribution of eNOS Glu298Asp polymorphism had no significant differ-ence between different blood pressure groups and gender groups,but there was a significant difference between dif-ferent age groups,diastolic blood pressure groups or BMI groups(P<0.05).Asp/Asp genotype significantly increased the risk of hypertension in individuals with serum TC above 5.4 mmol/L(P=0.03,OR=52.65).eNOS Glu298Asp polymorphism and serum lipid could synergis-tically modulate the blood pressure.eNOS Asp/Asp geno-type could significantly increase the risk of hypertension in individuals with serum TC over 5.4 mmol/L.eNOS Glu298Asp in combination with serum TC could be used to predict the risk of hypertension.

关键词： endothelial cells nitric oxide synthase type III hypertension serum lipid blood glucose

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