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简介：ScutellariaeRadix(SR),therootofScutellariabaicalensisGeorgi,isusedasanantipyreticdrugandhasbeendemonstratedtohaveanti-inflammatoryactivity.SRisdividedintotwospecifications,'KuQin'(KQ)and'ZiQin'(ZQ),foruseagainstdifferentsymptoms(upperenergizerheatorlowerportionofthetripleenergizer),accordingtothetheoryoftraditionalChinesemedicine(TCM).However,differencesintheefficaciesofthesetwospecificationshavenotbeendetermined.Inthepresentstudy,weaimedtocharacterizethedifferencesintheanti-inflammatoryactivitiesbetweenKQandZQandtoexplorehowtheirdifferencesaremanifestedinlipopolysaccharide(LPS)-inducedmacrophages.Ourresultsshowedthat,inRAW264.7cells(amousemacrophagecelllinederivedfromascites),KQandZQdisplayedanti-inflammatoryeffectsbyinhibitingthereleaseofnitricoxide(NO),inducibleNOS(iNOS),andnuclearfactor-κB(NF-κB)inadose-dependentmannerwithoutdistinction.InNR8383cells(aratalveolarmacrophagecellline),KQandZQdisplayedsimilareffectsonNO,iNOS,andNF-κBasseeninRAW264.7cells,butKQshowedahigherinhibitionrateforNOandiNOSthanthatshownbyZQatthesameconcentration.TheseresultsindicatedthatthereweredifferencesinefficacybetweenKQandZQintreatinglunginflammation.OurfindingsprovidedanexperimentalevidencesupportingthedifferentusesofKQandZQinclinic,asnotedinancientherbalrecords.

简介：AbstractBackground:Cardiac remodeling after acute myocardial infarction (AMI) is an important process. The present study aimed to assess the protective effects of astaxanthin (ASX) on cardiac remodeling after AMI.Methods:The study was conducted between April and September 2018. To create a rat AMI model, rats were anesthetized, and the left anterior descending coronary artery was ligated. The rats in the ASX group received 10 mg·kg-1·day-1 ASX by gavage for 28 days. On the 1st day after AMI, but before ASX administration, six rats from each group were sacrificed to evaluate changes in the heart function and peripheral blood (PB) levels of inflammatory factors. On the 7th day after AMI, eight rats from each group were sacrificed to evaluate the PB levels of inflammatory factors and the M2 macrophage count using both immunofluorescence (IF) and flow cytometry (FC). The remaining rats were observed for 28 days. Cardiac function was examined using echocardiography. The inflammatory factors, namely, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-10, were assessed using enzyme-linked immunosorbent assay. The heart weight/body weight (BW), and lung weight (LW)/BW ratios were calculated, and myocardial fibrosis in the form of collagen volume fraction was measured using Masson trichrome staining. Hematoxylin and eosin (H&E) staining was used to determine the myocardial infarct size (MIS), and TdT-mediated dUTP nick-end labeling staining was used to analyze the myocardial apoptosis index. The levels of apoptosis-related protein, type I/III collagen, transforming growth factor β1 (TGF-β1), metalloproteinase 9 (MMP9), and caspase 3 were assessed by Western blotting. Unpaired t-test, one-way analysis of variance, and non-parametric Mann-Whitney test were used to analyze the data.Results:On day 1, cardiac function was worse in the ASX group than in the sham group (left ventricular end-systolic diameter [LVIDs]: 0.72 ± 0.08 vs. 0.22 ± 0.06 cm, t= -11.38; left ventricular end-diastolic diameter [LVIDd]: 0.89 ± 0.09 vs. 0.48 ± 0.05 cm, t= -9.42; end-systolic volume [ESV]: 0.80 [0.62, 0.94] vs. 0.04 [0.03, 0.05] mL, Z = -2.89; end-diastolic volume [EDV]: 1.39 [1.03, 1.49] vs. 0.28 [0.22, 0.32] mL, Z = -2.88; ejection fraction [EF]: 0.40 ± 0.04 vs. 0.86 ± 0.05, t= 10.00; left ventricular fractional shortening [FS] rate: 0.19 [0.18, 0.20] %FS vs. 0.51 [0.44, 0.58] %FS, Z = -2.88, all P < 0.01; n = 6). The levels of inflammatory factors significantly increased (TNF-α: 197.60 [133.89, 237.94] vs. 50.48 [47.21 57.10] pg/mL, Z= -2.88; IL-1β: 175.23 [160.74, 215.09] vs. 17.78 [16.83, 19.56] pg/mL, Z = -2.88; IL-10: 67.64 [58.90, 71.46] vs. 12.33 [11.64, 13.98] pg/mL, Z = -2.88, all P < 0.01; n = 6). On day 7, the levels of TNF-α and IL-1β were markedly lower in the ASX group than in the AMI group (TNF-α: 71.70 [68.60, 76.00] vs. 118.07 [106.92, 169.08] pg/mL, F = 42.64; IL-1β: 59.90 [50.83, 73.78] vs. 151.60 [108.4, 198.36] pg/mL, F = 44.35, all P < 0.01, n = 8). Conversely, IL-10 levels significantly increased (141.84 [118.98, 158.36] vs. 52.96 [42.68, 74.52] pg/mL, F= 126.67, P < 0.01, n = 8). The M2 macrophage count significantly increased (2891.42 ± 211.29 vs. 1583.38 ± 162.22, F= 274.35, P < 0.01 by immunofluorescence test; 0.96 ± 0.18 vs. 0.36 ± 0.05, F= 46.24, P < 0.05 by flowcytometry test). On day 28, cardiac function was better in the ASX group than in the AMI group (LVIDs: 0.50 [0.41, 0.56] vs. 0.64 [0.56, 0.74] cm, Z = -3.60; LVIDd: 0.70 [0.60, 0.76] vs. 0.80 [0.74 0.88] cm, Z = -2.96; ESV: 0.24 [0.18, 0.45] vs. 0.58 [0.44, 0.89] mL, Z = -3.62; EDV: 0.76 [0.44, 1.04] vs. 1.25 [0.82, 1.46] mL, Z = -2.54; EF: 0.60 ± 0.08 vs. 0.50 ± 0.12, F= 160.48; % FS: 0.29 [0.24, 0.31] vs. 0.20 [0.17, 0.21], Z = -4.43, all P < 0.01; n = 16). The MIS and LW/BW ratio were markedly lower in the ASX group than in the AMI group (myocardial infarct size: 32.50 ± 1.37 vs. 50.90 ± 1.73, t = 23.63, P < 0.01, n = 8; LW/BW: 1.81 ± 0.15 vs. 2.17 ± 0.37, t= 3.66, P = 0.01, n = 16). The CVF was significantly lower in the ASX group than in the AMI group: 12.88 ± 2.53 vs. 28.92 ± 3.31, t = 10.89, P < 0.01, n = 8. The expression of caspase 3, TGF-β1, MMP9, and type I/III collagen was lower in the ASX group than in the AMI group (caspase 3: 0.38 ± 0.06 vs. 0.66 ± 0.04, t= 8.28; TGF-β1: 0.37 ± 0.04 vs. 0.62 ± 0.07, t = 6.39; MMP9: 0.20 ± 0.06 vs. 0.40 ± 0.06, t = 4.62; type I collagen: 0.42 ± 0.09 vs. 0.74 ± 0.07, t = 5.73; type III collagen: 0.13 ± 0.02 vs. 0.74 ± 0.07, t = 4.32, all P < 0.01; n = 4).Conclusions:ASX treatment after AMI may promote M2 macrophages and effectively attenuate cardiac remodeling by inhibiting inflammation and reducing myocardial fibrosis.

简介：Recently,thehumancochleahasbeenshowntocontainnumerousresidentmacrophagesundersteady-state.Themacrophagesaccumulateinthestriavascularis,amongtheauditorynerves,andarealsospottedinthehumanorganofCorti.ThesemacrophagesmayprocessantigensreachingthecochleabyinvasionofpathogensandinsertionofCIelectrode.Thus,macrophagesexecuteaninnate,andpossiblyanadaptiveimmunity.Here,wedescribethemolecularmarkersCD4andCD8ofTcells,macrophagemarkersMHCⅡandCD11b,aswellasthemicroglialmarkersTEME119andP2Y12,inthehumancochlea.Immunohistochemistryandtheadvantageoussuper-resolutionstructuredilluminationmicroscopy(SR-SIM)wereusedinthestudy.CD4~+andCD8~+cellswerefoundinthehumancochleae.Theywereseeninthemodiolusinasubstantialnumberadjacenttothevessels,intheperipheralregionoftheRosenthal’scanal,andoccasionallyinthespiralligament.Whilethereareasurprisinglylargenumberofmacrophagesinthestriavascularisaswellasbetweentheauditoryneurons,CD4~+andCD8~+cellsarehardlyseenintheseareas,andneitherareseenintheorganofCorti.Inthemodiolus,macrophages,CD4~+andCD8~+cellsappearedofteninclusters.InteractionbetweenthesedifferentcellswaseasilyobservedwithSR-SIM,showingcloselyplacedcellbodies,andtheprocessesfrommacrophagesreachingoutandtouchingthelymphocytes.OtherwisetheCD4~+andCD8~+cellsinhumancochleartissuearediscretelyscattered.Thepossiblerolesoftheseimmunecellsarespeculated.

简介：AbstractBackground:Previous evidence suggests inflammation may be a double-edged sword with cancer-promoting and cancer suppressing function. In this study, we explore the impact of local and systemic inflammation on cancer growth.Methods:Female BALB/C mice were subcutaneously implanted with foreign body (plastic plates) to build up a local inflammation and intraperitoneally injected with PolyIC or lipopolysaccharides (LPS) to build up a systemic inflammation, followed by subcutaneous injection of 5 × 105 colon cancer cells. Immunohistochemistry and enzyme linked immunosorbent assay were utilized to detect the Ki67 and interleukin (IL) 6, IL-1β, and monocyte chemoattractant protein-1 expression in the tumor tissues and serum, respectively. The distributions of immune cells and expression of toll-like receptors (TLRs) were evaluated by flow cytometry (FCM) and quantitative real time-polymerase chain reaction.Results:The results showed that local inflammation induced by foreign body implantation suppressed tumor growth with decreased tumor weight (P = 0.001), volume (P = 0.004) and Ki67 index (P < 0.001). Compared with the control group, myeloid-derived suppressive cells sharply decreased (P = 0.040), while CD4+ T cells slightly increased in the tumor tissues of the group of foreign body-induced local inflammation (P = 0.035). Moreover, the number of M1 macrophages (P = 0.040) and expression of TLRs, especially TLR3 (P < 0.001) and TLR4 (P < 0.001), were significantly up-regulated in the foreign body group. Contrarily, tumor growth was significantly promoted in LPS or PolyIC-induced systemic inflammation (P = 0.009 and 0.006). FCM results showed M1 type macrophages (P = 0.017 and 0.006) and CD8+ T cells (P = 0.031 and 0.023) were decreased, while M2 type macrophages (P = 0.002 and 0.007) were significantly increased in tumor microenvironment of LPS or PolyIC-induced systemic inflammation group. In addition, the decreased expression of TLRs was detected in LPS or PolyIC group.Conclusions:The foreign body-induced local inflammation inhibited tumor growth, while LPS or PolyIC-induced systemic inflammation promoted tumor growth. The results suggested that the different outcomes of tumor growth might be attributed to the infiltration of anti-tumor or pro-tumor immune cells, especially M1 or M2 type macrophages into tumor microenvironment.

简介：AnovelPleurotusnebrodensispolysaccharide(PN-S)waspurifiedandcharacterized,anditsimmune-stimulatingactivitywasevaluatedinRAW264.7macrophages.PN-SinducedtheproliferationofRAW264.7cellsinadose-dependentmanner,asdeterminedbytheMTTassay.AfterexposuretoPN-S,thephagocytosisofthemacrophageswassignificantlyimproved,withremarkablechangesinmorphologybeingobserved.FlowcytometricanalysisdemonstratedthatPN-SpromotedRAW264.7cellstoprogressthroughSandG2/Mphases.PN-Streatmentenhancedtheproductionsofinterleukin-6(IL-6),nitricoxide(NO),interferongamma(INF-γ),andtumornecrosisfactor-α(TNF-α)inthemacrophages,withup-regulationofmRNAexpressionsofinterleukin-6(IL-6),induciblenitricoxidesynthase(iNOS),interferongamma(INF-γ)andtumornecrosisfactor-α(TNF-α)beingobservedinadose-dependentmanner,asmeasuredbyqRT-PCR.Inconclusion,theseresultssuggestthatthepurifiedPN-Scanimproveimmunitybyactivatingmacrophages.

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简介：Themoditicationoftumorcellsoreffcorcellsusingcytokinegenesasastrategytoenhancehostantitumorimmunityhasbeenstudiedintensivelyoverthe

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简介：AbstractBackground:Macrophages play an important role in renal ischemia reperfusion injury, but the functional changes of macrophages under hypoxia/reoxygenation and the related mechanism are unclear and need to be further clarified.Methods:The effects of hypoxia/reoxygenation on functional characteristics of RAW264.7 macrophages were analyzed through the protein expression detection of pro-inflammatory factors TNF-α and CD80, anti-inflammatory factors ARG-1 and CD206. The functional implications of C-X3-C motif chemokine receptor 1(CX3CR1) down-regulation in hypoxic macrophages were explored using small interfering RNA technology. Significance was assessed by the parametric t-test or nonparametric Mann-Whitney test for two group comparisons, and a one-way ANOVA or the Kruskal-Wallis test for multiple group comparisons.Results:Hypoxia/reoxygenation significantly increased the protein expression of M1-related pro-inflammatory factors TNF-α, CD80 and chemokine C-X3-C motif chemokine ligand 1 (CX3CL1)/CX3CR1 and inhibited the protein expression of M2-related anti-inflammatory factors ARG-1 and CD206 in a time-dependent manner in RAW264.7 cells. However, the silencing of CX3CR1 in RAW264.7 cells using specific CX3CR1-siRNA, significantly attenuated the increase in protein expression of TNF-α (P < 0.05) and CD80 (P < 0.01) and the inhibition of ARG-1 (P < 0.01) and CD206 (P < 0.01) induced by hypoxia/reoxygenation. In addition, we also found that hypoxia/reoxygenation could significantly enhance the migration (2.2-fold, P < 0.01) and adhesion capacity (1.5-fold, P < 0.01) of RAW264.7 macrophages compared with the control group, and CX3CR1-siRNA had an inhibitory role (40% and 20% reduction, respectively). For elucidating the mechanism, we showed that the phosphorylation levels of ERK (P < 0.01) and the p65 subunit of NF-κB (P < 0.01) of the RAW264.7 cells in the hypoxic/reoxygenation group were significantly increased, which could be attenuated by down-regulation of CX3CR1 expression (P < 0.01, both). ERK inhibitors also significantly blocked the effects of hypoxic/reoxygenation on the protein expression of M1-related pro-inflammatory factors TNF-α, CD80 and M2-related anti-inflammatory factors ARG-1 and CD206. Moreover, we found that conditioned medium from polarized M1 macrophages induced by hypoxia/reoxygenation, notably increased the degree of apoptosis of hypoxia/reoxygenation-induced TCMK-1 cells, and promoted the protein expression of pro-apoptotic proteins bax (P < 0.01) and cleaved-caspase 3 (P < 0.01) and inhibited the expression of anti-apoptotic protein bcl-2 (P < 0.01), but silencing CX3CR1 in macrophages had a protective role. Finally, we also found that the secretion of soluble CX3CL1 in RAW264.7 macrophages under hypoxia/reoxygenation was significantly increased.Conclusions:The findings suggest that hypoxia/reoxygenation could promote M1 polarization, cell migration, and adhesion of macrophages, and that polarized macrophages induce further apoptosis of hypoxic renal tubular epithelial cells by regulating of CX3CL1/CX3CR1 signaling pathway.

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简介：cAMPmediatedsignalingmayplayasuppressiveroleinimmuneresponse.WepreviouslyfoundthatthecAMP-elevators(CTxand8-Br-cAMP)inhibitedIL-12,IL-la,IL-6geneexpression,butincreasedthetranscriptionallevelsofIL-10andIL-1RainLPS-treatedmurineperitonealmacrophages.ThepresentstudyexaminedapossiblemolecularmechanisminvolvedincAMPelevators-inducedinhibitionofIL-12p40expressioninresponsetoLPS.OurdatademonstratedthatcAMPelevatorsdownregulatedIL-12p40mRNAexpressionandIL-12pT0productioninmurineperitonealmacrophages.SubsequentstudiesrevealedthatcAMP-elevatorsblockedphosphorylationofp38MAPK,butdidnotaffecttheactivityofNF-κBbindingtoIL-12promoter(-136/-112).ThisisthefirstreportthatcAMPelevatorsinhibitLPS-inducedIL-12productionbyamechanismthatisassociated,atleastinpart,withp38-dependentinhibitionbycAMPsignalingpathways.

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简介：我们表明白氨酸应答的规章的蛋白质(rLrp)禁止的那Mycobacterium肺结核recombinantlipopolysaccharide(LPS)导致了肿瘤坏死因素alpha(TNF-α；)，interleukin-6，和interleukin-12生产和块原子受体的抄写因素NF-κ的子单元的原子translocation；B(原子factor-kappaB)。而且，rLrp稀释导致LPS的DNA绑定和NF-κ；Btranscriptional活动，被禁止的Iκ的降级伴随，Bα；并且在NF-κ的原子translocation的作为结果的减少；Bp65子单元。RLrp防碍TNF联系受体的因素的导致LPS的聚类6并且与interleukin-1联系受体的kinase1绑定到TAK1。而且，rLrp没稀释proinflammatorycytokines或CD86和interferon-gamma在像使用费的受体的巨噬细胞导致的主要histocompatibility建筑群class-II的表示2删除(TLR2−/−)老鼠并且在蛋白质kinase，b(Akt)弄空老鼠房间，显示rLrp的禁止的效果依赖于phosphatidylinositol3-OHkinase(PI3K)的调停TLR2的激活/Akt小径。RLrp能也由刺激PI3K，Akt，和肝糖synthasekinase的快速的phosphorylation激活PI3K/Akt小径在巨噬细胞的3贝它。另外，在rLrp的N终点的19氨基酸残余决心重要、要求因为禁止的效果由TLR2调停了。rLrp的这19氨基酸的禁止的功能提起模仿的禁止的肽能被用来处于发信号的延长TLR在有有害效果的状况限制天生的有免疫力的回答的可能性。我们的学习提供新卓见进调停TLR2的PI3K/Akt小径由保证有势力的短暂表示的禁止的机制煽动性的调停人。

简介：Brucella流产胎是zoonotic在ruminants和人引起brucelosis的克否定的病原体。像使用费的受体(TLR)认出Brucella流产胎并且开始影响天生、适应的免疫的介绍抗原的房间活动。在这研究，我们集中了于recombinantBrucella房间表面蛋白质31(rBCSP31)在老鼠巨噬细胞上决定它的效果。我们表明的结果rBCSP31导致了TNF-α；，IL-6和IL-12p40生产，由刺激p38和JNK和抄写因素NF-κ的激活的快速的phosphorylation取决于激活mitogen的蛋白质kinases(MAPK)的激活，在巨噬细胞的B。另外，连续暴露(>；24h)RAW264.7，到rBCSP31的房间显著地提高了IFN-γ；MHC-II的导致的表示和介绍rBCSP31肽给CD4+T房间的能力。而且，我们发现rBCSP31能与TLR2和TLR4交往。由从TLR2−/−和TLR4−/−老鼠的巨噬细胞的导致rBCSP31的cytokine生产从NF-κ的C57BL/6巨噬细胞，和激活是比那低的；B和MAPK从TLR2−/−和TLR4−/−老鼠在巨噬细胞被稀释。另外，从与rBCSP31使免疫的C57BL/6老鼠的CD4+T房间生产了IFN-γ的高水平；并且IL-2与从TLR2−/−和TLR4−/−老鼠的CD4+T房间相比。从使免疫的C57BL/6老鼠的巨噬细胞从TLR2−/−和TLR4−/−老鼠比那些生产了IL-12p40的高水平。而且，有rBCSP31的免疫在B以后比在TLR2−/−和TLR4−/−老鼠在C57BL/6老鼠提供了更好的保护。流产胎2308挑战。这些结果显示rBCSP31是导致cytokine生产的TLR2和TLR4收缩筋，upregulates巨噬细胞功能并且导致Th1有免疫力的反应。

简介：AbstractBackground:Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.Methods:We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditions in vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.Results:The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20 vs. 0.44 ± 0.08, t = 6.67, P < 0.05), while the expression of p62 was decreased (0.77 ± 0.04 vs. 0.95 ± 0.10, t = 2.90, P < 0.05), and PI3K (0.40 ± 0.06 vs. 0.63 ± 0.10, t = 3.42, P < 0.05) and p-Akt/Akt ratio was also decreased (0.39 ± 0.02 vs. 0.58 ± 0.03, t = 9.13, P < 0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68 ± 0.14 vs. 1.27 ± 0.20, t = 4.12, P < 0.05), up-regulated p62 expression (1.10 ± 0.20 vs. 0.77 ± 0.04, t = 2.80, P < 0.05), and up-regulated PI3K (0.54 ± 0.05 vs. 0.40 ± 0.06, t = 3.11, P < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05 vs. 0.39 ± 0.02, t = 9.13, P < 0.05). A whole-genome microarray assay screened the differentially expressed gene HO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration of HO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway.Conclusions:Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstances in vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.