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Effect of cholecystokinin octapeptide on diacylglyceroI-PKC signaling pathway in rat pulmonary interstitial macrophages stimulated by lipopolysaccharide

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Acta Pharmacologica Sinica 2005年第12期26卷 1497-1504页

作者： Shun-jiang XU~2 Wei-juan GAO Bin CONG~3 Chun-ling MA Shu-jin LI Yi-ling LING Zhen-yong GU Yu-xia YAO Department of Pathophysiology,Hebei Medical University,Shijiazhuang 050017,China the Hebei Province Center for Disease Prevention and Control,Shijiazhuang 050021,China. Department of Pathophysiology Hebei Medical University Shijiazhuang China

Aim:To investigate the effect of cholecystokinin octapeptide(CCK-8)on thediacylglycerol-protein kinase C(DAG-PKC)signaling pathway in rat pulmonaryinterstitial macrophages(PIM)stimulated by lipopolysaccaride(LPS).Methods:The PIM from rat lung tissues were isolated using the collagenase digestionmethod combined with alveolar lavage and pulmonary vessel perfusion.DAGcontent and PKC activity were measured by radioenzymatic assay.The transloca-tion of PKCζ was determined by semi-quantitative immunoblot analysis.Results:CCK-8,at high concentrations(1×10-6-1×10-5mol/L),decreased DAG content andinhibited PKC activity and PKCζ translocation compared with that in rat restingPIM of a control group(P<0.01).LPS increased DAG content,and promoted PKCactivity and PKC~ translocation(P<0.01).CCK-8 decreased LPS-induced DAGcontent and inhibited LPS-induced PKC activity and PKCζ translocation signifi-cantly at 1×10-8-1×10-5mol/L(P<0.01).This inhibitory effect of CCK-8 could beabrogated partly by proglumide(non-selective CCK receptor antagonist),CR-1409(selective CCK-A receptor antagonist),and CR-2945(selective CCK-Breceptor antagonist)in a concentration-dependent manner(P<0.01).Conclusion:CCK-8 was a negative modulator of the DAG-PKC signaling pathway in rat restingPIM,which is very important for maintaining body homeostasis.It significantlyinhibited LPS-induced DAG content,PKC activity and PKCζ translocation in aconcentration-dependent manner.The CCK receptor,especially the CCK-Areceptor,might play a major role in this process.

关键词： cholecystokinin protein kinase C translocation lung macrophages lipopolysaccharides

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Effect of Tumor Necrosis Factor-α on Acyl Coenzyme A: Cholesteryl Acyltransferase Activity and ACAT1 Gene Expression in THP-1 macrophages

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第2期27卷 170-172页

作者：何平成蓓王毅王洪星 Department of Gerontology Union Hospital Tongji Medical College Huazhong University of Science and Technology Department of Cardiology the First People's Hospital Shanghai Jiao Tong University Department of Molecular and Cell Biology Zeneca Pharmaceuticals

In order to explore the effect and mechanisms of tumor necrosis factor-α （TNF-α） on the activity of the acyl coenzyme A： cholesteryl acyltransferase （ACAT）, THP-I monocytes were cul- tured and induced to differentiate into macrophages with phorbol ester. TNF-α （60 ng/mL） was added at different time points into the macrophage-containing medium and the ACAT enzyme activity was measured by quantifying the incorporation of [1-^14C] oleoyl CoA into cholesteryl esters. The expression of ACAT-1 protein and mRNA was respectively detected by Western blotting and RT-PCR in THP-1 macrophages 24 h after treatment with TNF-α （60 ng/mL）. The results indicated that ACAT activity in THP-I macrophages treated with TNF-α was increased in a time-dependent manner. The expression levels of ACAT-1 protein and mRNA were significantly increased in THP-I macrophages after treatment with TNF-α （P〈0.05）. It was suggested that TNF-α could increase the activity of ACAT in THP-1 macrophages by up-regulating the expression of ACAT-1 gene.

关键词： acyl coenzyme A： cholesteryl acyltransferase tumor necrosis factor-α macrophages cholesterol atherosclerosis

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Effects of Rapamycin on the Differentiation and Function of macrophages In Vitro

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Agricultural Sciences in China 2009年第5期8卷 620-627页

作者： SHEN Hong WU Guo-juan LIU Guang-wei MA Hai-xia LIAO An-yan ZHAO Yong Department of Animal Science and Technology Beijing University of Agriculture Beijing 102206 P.R.China State Key Laboratory of Biomembrane and Membrane Biotechnology/Institute of Zoology Chinese Academy of Sciences Beijing 100101 P.R. China The Third Hospital Affiliated to Beijing University of Chinese Medicine Beijing 100029 P.R.China

macrophages have been generated from bone marrow progenitor cells （BMPCs） with macrophage colony-stimulating factor （M-CSF）. Rapamycin （rapa） is an immunosuppressive drug, which could prevent renal graft rejection and inhibit tumor growth to yield antiproliferative activity in a variety of malignancies. However, the direct effect of rapa on the development of macrophages is unknown. In this study, we explored the direct effect of rapa on the differentiation and function of macrophages differentiated from mouse BMPCs in vitro in the presence of M-CSF. The experimental data showed that rapa prevented the differentiation of macrophagcs by down-regulating CD80 and CD86 expression but upregulating F4/80 expression, as well as by reducing the capacity of differentiated macrophages to stimulate lymphocyte proliferation in the allogeneic mixed lymphocyte reaction. Furthermore, the phagocytic capacity of differentiated macrophages was significantly reduced by rapa. Therefore, rapa may directly inhibit macrophage differentiation and function, which may have been one of the major targets for rapa to mediate its immunosuppressive properties.

关键词： function macrophages rapamycin differentiation

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Effect of HCMV IE1 Protein on Cytokines Secretion and Apoptosis of macrophages

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Chinese Journal of Cancer Research 2008年第1期20卷 12-16页

作者：陈琳万艳平陈熙刘安元朱翠明余敏君曹清香 Institute of Pathogenic Biology University of South China

Objective： To investigate the effect of human cytomegalovirus （HCMV） IE1 protein on the secretory activity and apoptosis of macrophages. Methods： The eukaryotic expression vector pEGFP-C1/IE1 was used to transfect THP-1-macrophages. 48 h after transfection, the expression and localization of GFP or GFP-IE1 was observed under fluorescent microscope. The levels of IL-1β and TNF-α in the culture media were examined by ELISA, and the mRNA expression of them was analyzed by RT-PCR. Cell undergoing apoptosis were determined by flow cytometry using the propidium iodide （PI） staining method. The data were analyzed by SPSSI3.0. Results： As observed under fluorescent microscope, the expressions of GFP-IEI and GFP by plasmid pEGFP-C1/IE1 or pEGFP-C1 in THP-1-macrophages could be found in nuclei or whole cells. Conclusion： As demonstrated by RT-PCR and ELISA, mRNA and protein expressions of IL-1β and TNF-α and promotes apoptosis in THP-1-macrophages.

关键词： Human cytomegalovirus （HCMV） IE1 macrophages IL-1β TNF-α Apoptosis

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The effect of Helicobacter pylori infection on expression of macrophage migration inhibitory factor by T cells and macrophages in gastric mucosa

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Chinese Medical Journal 2005年第14期118卷 1201-1205页

作者： HEXing-xiang YANGJun ZHENGXue-ling DINGYuan-wei SHENQing-yan LIUWei ZHAOYing-heng DepartmentofGastroenterology theSecondAffiliatedHospitalofGuangzhouMedicalCollegeGuangzhou510260China DepartmentofEpidemiology RoswellParkCancerInstituteElmandCarltonStreetsBuffaloNewYork14263USA

Background Macrophage migration inhibitory factor (MIF) plays a pivotal role in inflammatory and immune-mediated diseases. However, its molecular function and role in gastrointestinal diseases has rarely been studied and thus warrants an in-depth investigation. This study was designed and conducted to determine MIF expression in Helicobacter pylori (H. pylori)-induced gastritis and the effect of H. pylori on MIF expression in monocytes in vitro. Methods Gastric specimens of 62 patients with chronic gastritis were obtained through endoscopic biopsies. Both gastric antrum and body were examined for histopathologic changes. Positive H. pylori was determined through rapid urease test and histopathological examination. A patient was classified as H. pylori positive if both tests showed positive results. The updated Sydney System was employed to assess the severity and activity of gastric inflammation. Double immunoassaying for MIF/T-cells (CD45RO) and MIF/macrophage (KP1), as well as in situ hybridization for the expression of MIF mRNA were used for the current analysis. THP-1, a monocyte cell line, was co-incubated with H. pylori strains (ATCC26695) and subsequently examined for the expression of MIF protein and mRNA by enzyme linked immunosorbent assay and retrospective transcription-polymerase chain reaction, respectively. Results Among 62 patients with chronic gastritis, significant increase in total T-cells, MIF+ T-cells, total macrophages, MIF+ macrophages and MIF mRNA+ cells was observed in 42 H. pylori positive patients compared to H. pylori negative patients. Moreover, the increase of the MIF mRNA+ cells was highly correlated with the severity of the disease(number of MIF mRNA+cells/mm2, mild: 2834±382, moderate: 3569±123, severe: 3881±118, P<0.01). In vitro results showed that the expression of MIF protein and mRNA in monocytes was significantly increased after incubation with H. pylori strains.Conclusions Overexpression of MIF is common in H. pylori-induced gastric inflammation, which suggests MIF may play

关键词： macrophage migration inhibitory factor Helicobacter pylori gastric inflammation T cells macrophages

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Hepatitis B Virus X Protein Up-regulates TNF-α and IL-1β Secretion of macrophages

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Chinese Journal of Cancer Research 2007年第3期19卷 206-209页

作者：陈春莲吴移谋王静朱翠明刘安元尹卫国万艳平 Institute of Pathogenic Biology University of South China

Objective： To provide the experimental basis for further studying the molecular transformation mechanism of Hepatitis B virus （HBV） X protein （HBx） on hepatocellular carcinoma. Methods： Reconstructed plasmid pcDNA3.1（＋）-HBx was transfected into THP-1 macrophages. Expression of HBx was assayed in macrophages lysate by Western-blotting, and TNF-α and IL-1β contents were detected respectively by ELISA. All the data were analyzed by SPSS13.0. Results： In THP-lmacrophages, the pcDNA3.1（＋）-HBx plasmid expressed HBx with a molecular weight of about 17 KDa demonstrated by Western-blotting. The secreted TNF-α and IL-1β from macrophages were determined by ELISA, the results from analysis of all groups showed as following： control group was different from LPS group and pcDNA3.1（＋） group （P〈0.01）, and so was pcDNA3.1（＋）-HBx group; but there was no obvious difference between pcDNA3.1（＋） group and LPS group （P〉0.05）, all of which indicated that transient overexpression of HBx enhanced LPS-induced production of TNF-α and IL-1β by macrophages. Conclusion： Transient overexpression of HBx up-regulates LPS-induced TNF-t~ and IL-113 secretion of macrophages.

关键词： HBV X protein macrophages TNF-α IL-1β

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Role of hypoxia inducible factor 1α in cobalt nanoparticle induced cytotoxicity of human THP-1 macrophages

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Biomaterials Translational 2021年第2期2卷 143-150页

作者： Wendy Rachel Francis Zhao Liu Sian E Owens Xiao Wang Huaming Xue Alex Lord Venkateswarlu Kanamarlapudi Zhidao Xia Centre for Nanohealth Swansea University Medical SchoolSwanseaUK Department of Orthopaedic Surgery Orthopaedic InstituteThe First Affiliated Hospital of Soochow UniversitySuzhouJiangsu ProvinceChina Department of Orthopaedic Surgery Johns Hopkins School of MedicineBaltimoreMDUSA Department of Orthopaedics Yangpu Hospital affiliated to Tongji University School of MedicineShanghaiChina.

Cobalt is one of the main components of metal hip prostheses and cobalt nanoparticles(CoNPs)produced from wear cause inflammation,bone lyses and cytotoxicity at high concentrations.Cobalt ions mimic hypoxia in the presence of normal oxygen levels,and activate hypoxic signalling by stabilising hypoxia inducible transcription factor 1α(HIF1α).This study aimed to assess in vitro the functional role of HIF1αin CoNP induced cellular cytotoxicity.HIF1α,lysosomal pH,tumour necrosis factorαand interleukin 1βexpression were analysed in THP-1 macrophages treated with CoNP(0,10 and 100μg/mL).HIF1αknock out assays were performed using small interfering RNA to assess the role of HIF1αin CoNP-induced cytotoxicity.Increasing CoNP concentration increased lysosomal activity and acidity in THP-1 macrophages.Higher doses of CoNP significantly reduced cell viability,stimulated caspase 3 activity and apoptosis.Reducing HIF1αactivity increased the pro-inflammatory activity of tumour necrosis factorαand interleukin 1β,but had no significant impact on cellular cytotoxicity.This suggests that whilst CoNP promotes cytotoxicity and cellular inflammation,the apoptotic mechanism is not dependent on HIF1α.

关键词： cobalt nanoparticle cytotoxicity hypoxia inducible factor macrophages TNFα

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Tumor tissue stiffness regulates macrophages differentiation and functions

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医用生物力学 2013年第S1期28卷 132-133页

作者： Yi Zhang Huafeng Zhang Bo Huang Department of Immunology Chinese Academy of Medical Sciences

Background&Objective Tumor-associated macrophages(TAM)are induced by many cytokines,such as IL-4,IL-10,IL-13,and are considered to be of the M2 phenotype,which provides an immunosuppressive microenvironment for tumor growth.However,biomechanistssuggest that cells in the tumor microenvironment are affected not only by chemical signals but also mechanical factors,of which the stiffness of tissues is a major determinant.Compared with normal tissues,including the peri-tumor tissues,tumor tissues are more rigid.Heretofore,the influence,on the differentiation and function of macrophages,of the differences in such physical aspect between

关键词： macrophages stiffness cytokines phenotype rigid determinant cytometry tumorigenesis nude inhibited

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Paeoniflorin prevents hepatic fibrosis of Schistosomiasis japonica by inhibiting TGF-β1 production from macrophages in mice

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Frontiers of Medicine 2008年第2期2卷 154-165页

作者： Deyong CHU Conglei LI Qiang WU Jilong SHEN Institute of Zoonoses of Anhui Medical University and the Key Laboratory of Gene Resource Utilization for Severe Diseases Anhui Medical UniversityHefei 230032China Department of Pathology Anhui Medical UniversityHefei 230032China

In order to investigate the effect of paeoniflorin(PAE)on hepatic fibrosis of mice with Schistosomiasis japonica in vivo and in vitro,a model of hepatic fibrosis caused by schistosomiasis was established in mice infected with cercariae of Schistosoma japonicum.Then,PAE was orally administered before and after praziquantel treat-ment and both therapeutics were given simultaneously at different time points after the infection.The concentra-tion of serum hyaluronic acid(HA)was determined by radioimmunoassay(RIA).Hepatic granuloma and fib-rosis were evaluated via HE and Masson staining.The expression of α-smooth muscle actin(α-SMA),transform-ing growth factor β1(TGF-β1)and collagen I(Col I)protein was detected by immunohistochemistry.The effect of soluble egg antigen(SEA)and PAE on the pro-duction of TGF-β1 from mouse peritoneal macrophages(PMQs)was investigated by RT-PCR,Western blotting and ELISA.The effect of TGF-β1 in optimum macro-phage-conditioned medium(OPMCM)on the prolifera-tion of hepatic stellate cells(HSCs)and collagen secretion from HSCs with anti-TGF-β1 antibody was explored by MTT assay and ELISA.The results show that PAE could significantly reduce the concentration of serum HA,the size of egg granuloma,the severity of hepatic fibrosis and the expression of a-SMA,TGF-β1 and Col I protein in the pre-treatment group.However,in sim-or post-treatment group,PAE did not have any significant therapeutic effect.TGF-β1 could be secreted from PMQs stimulated by SEA.Meanwhile,the production of TGF-β1 from PMQs could be depressed significantly by PAE in a con-centration-dependent manner.TGF-β1 could promote the proliferation of HSCs and the secretion of collagens.In a word,PAE can prevent hepatic granuloma and fib-rosis caused by schistosomiasis japonica through the inhibition of the secretion of TGF-β1 from PMQs,the proliferation and activation of HSCs and the secretion of collagens from HSCs.

关键词： paeoniflorin Schistosomiasis japonica liver cirrhosis transforming growth factorβ1 macrophages hepatic stellate cell

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It takes two: potential therapies and insights involving microglia and macrophages in glioblastoma

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Neuroimmunology and Neuroinflammation 2018年第10期5卷 1-14页

作者： John Choi Nicholas Mai Christopher Jackson Zineb Belcaid Michael Lim Department of Neurosurgery Johns Hopkins School of Medicine Johns Hopkins University Baltimore MD 21231 USA

Microglia and macrophages, two myeloid cell lineages with different origins, make up the majority of immune cells present in glioblastoma (GBM). However, much of the literature does not distinguish between microglia and macrophages, despite a growing body of evidence that demonstrates key structural and functional differences between the cell types. Furthermore, the current M1/M2 paradigm used to sub-classify microglia and macrophages has proven to be incomplete at best, with the growing amount of in vivo and genomic data incompatible with this dichotomy. Finally, a number of studies have already established that in the setting of the GBM tumor microenvironment, both microglia and macrophages are complicit in tumor progression. This review highlights the differences between microglia and macrophages, particularly in the context of GBM, and discusses at length several potential therapeutic strategies made possible by understanding specific pro-tumor and anti-tumor pathways in these myeloid populations. Ultimately, investigating the differences between microglia and macrophages offers insight into the progression of GBM, its marked resistance to current immunotherapy regimens, and future directions for new treatment modalities.

关键词： Glioblastoma cancer immunotherapy myeloid microglia macrophages pro-tumor anti-tumor immunosuppression

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