Atherosis-associated lnc_000048 activates PKR to enhance STAT1-mediated polarization of THP-1 macrophages to M1 phenotype

Our previous study has demonstrated that lnc_000048 is upregulated in large-artery atherosclerotic stroke and promotes atherosclerosis in ApoE^(-/-)mice.However,little is known about the role of lnc_000048 in classically activated macrophage(M1)polarization.In this study,we established THP-1-derived testing state macrophages(M0),M1 macrophages,and alternately activated macrophages(M2).Real-time fluorescence quantitative PCR was used to verify the expression of marker genes and the expression of lnc_000048 in macrophages.Flow cytometry was used to detect phenotypic proteins(CD11b,CD38,CD80).We generated cell lines with lentivirus-mediated upregulation or downregulation of lnc_000048.Flow cytometry,western blot,and real-time fluorescence quantitative PCR results showed that down-regulation of lnc_000048 reduced M1 macrophage polarization and the inflammation response,while over-expression of lnc_000048 led to the opposite effect.Western blot results indicated that lnc_000048 enhanced the activation of the STAT1 pathway and mediated the M1 macrophage polarization.Moreover,catRAPID prediction,RNA-pull down,and mass spectrometry were used to identify and screen the protein kinase RNA-activated(PKR),then catRAPID and RPIseq were used to predict the binding ability of lnc_000048 to PKR.Immunofluorescence(IF)-RNA fluorescence in situ hybridization(FISH)double labeling was performed to verify the subcellular colocalization of lnc_000048 and PKR in the cytoplasm of M1 macrophage.We speculate that lnc_000048 may form stem-loop structure-specific binding and activate PKR by inducing its phosphorylation,leading to activation of STAT1 phosphorylation and thereby enhancing STAT1 pathway-mediated polarization of THP-1 macrophages to M1 and inflammatory factor expression.Taken together,these results reveal that the lnc_000048/PKR/STAT1 axis plays a crucial role in the polarization of M1 macrophages and may be a novel therapeutic target for atherosclerosis alleviation in stroke.

Macrophage Inflammatory Protein-1 Beta (MIP-1<i>β</i>) and Platelet Indices as Predictors of Spontaneous Bacterial Peritonitis<br>—MIP, MPV and PDW in SBP

Background/Aims: The objective of this study is to measure macrophage inflammatory protein one beta (MIP-1β), mean platelet volume (MPV) and platelet distribution width (PDW) to evaluate their usefulness in the diagnosis of spontaneous bacterial peritonitis (SBP) in cirrhotic patients. Materials and Methods: This study comprised 41 cirrhotic patients with ascites. MPV, PDW and MIP-1β were measured in serum and ascitic fluid. Results: A significant increase MPV, PDW, C-reactive Protein (CRP) and white blood cell was observed in SBP group compared to non SBP (P ≤ 0.001, P = 0 β was significantly in-creased in ascitic fluid in patients with SBP versus non SBP (P ≤ 0.001). At cutoff value of 8.3 fl MPV had 85.7% sensitivity and 75% specificity (AUC = 0.876) for diagnosis of SBP. At cutoff value of 15.4 PDW had 90.4% sensitivity and 55% specificity (AUC = 0.762). At cutoff value of 121.9 pg/ml MIP-1β in ascitic fluid had 76.1% sensitivity and 100% specificity (AUC = 0.881) for detecting SBP. Conclusion: MIP-1β and platelet indices are useful marker in the diagnosis of SBP in cirrhotic patients. Combined measurement of MIP-1β in serum and ascitic fluid had 100% sensitivity and specificity for diagnosis of SBP.

Effect of Mnk1 on lipopolysaccharide-induced inflammatory responses in macrophages

Objective:To investigate the effect of mitogen-activated protein kinase interaction serine kinase 1(Mnk1)gene deletion on lipopolysaccharide(LPS)-induced inflammatory response in mouse macrophages(Mφ)and the possible mechanism.Methods:Healthy male wildtype C57BL/6J(WT)and Mnk1 knockout(KO)mice were selected at 8-10 weeks of age and divided into WT+PBS,KO+PBS,WT+LPS and KO+LPS groups,and the serum levels of IL-1βwere measured by ELISA after 24 h intraperitoneal injection of PBS or LPS.The mRNA expression levels of IL-1βand Sprouty2(Spry2)in the spleen Mφwere measured by qRTPCR.Mφwas also extracted from the peritoneal cavity of two strains of mice for in vitro experiments to detect macrophage adhesion function and stimulated with equal volumes of LPS or PBS solution for 24 h,divided into WT+PBS group,KO+PBS group,WT+LPS group and KO+LPS group,and transfected with adenovirus expressing Spry2.qRT-PCR was used to detect the mRNA expression levels of LFA-1α,IL-1β,iNOS,CD206,Arg1 and Spry2 in Mφ.Mnk1,ERK1/2,P-ERK1/2,P-p38,P-JNK and Spry2 protein levels in Mφwere detected by western blot.Results:In the in vivo experiments,the concentration of IL-1βin the serum of the KO+LPS group was more significantly elevated than that of the WT+LPS group in mice injected intraperitoneally with LPS.The expression level of splenic MφIL-1βwas higher and the mRNA expression level of Spry2 was decreased in the KO+LPS group compared to the WT+LPS group.In the in vitro experiments,the mRNA expression levels of IL-1βand iNOS were elevated and those of CD206,Arg1 and Spry2 were decreased in the KO+LPS group compared with the WT+LPS group;the expression of LFA-1αwas not significantly different in the WT+PBS and WT+LPS groups,while the expression level of LFA-1αwas significantly increased in the KO+LPS group compared with the WT+LPS group.The results of the macrophage adhesion function assay showed that the adhesion rate of Mφin the KO group was increased at several time points compared to the WT group.After LPS stimulation,the expression of MφSpry2 decreased in Mnk1 KO group compared to WT group,while the expression of P-ERK1/2 increased compared to WT group.After Mφwas transfected with adenovirus overexpressing Spry2 and stimulated with LPS,MφSpry2 expression increased in the KO+AdSpry2 group and P-ERK1/2 expression decreased significantly compared to KO+AdGFP.Conclusion:Mnk1 knockdown enhances LPS-induced inflammatory responses in macrophages,and the mechanism may be related to the involvement of Spry2,a substrate of Mnk1,in regulating macrophage function.

The action mechanism by which C1q/tumor necrosis factor-related protein-6 alleviates cerebral ischemia/reperfusion injury in diabetic mice

Studies have shown that C1q/tumor necrosis factor-related protein-6 (CTRP6) can alleviate renal ischemia/reperfusion injury in mice. However, its role in the brain remains poorly understood. To investigate the role of CTRP6 in cerebral ischemia/reperfusion injury associated with diabetes mellitus, a diabetes mellitus mouse model of cerebral ischemia/reperfusion injury was established by occlusion of the middle cerebral artery. To overexpress CTRP6 in the brain, an adeno-associated virus carrying CTRP6 was injected into the lateral ventricle. The result was that oxygen injury and inflammation in brain tissue were clearly attenuated, and the number of neurons was greatly reduced. In vitro experiments showed that CTRP6 knockout exacerbated oxidative damage, inflammatory reaction, and apoptosis in cerebral cortical neurons in high glucose hypoxia-simulated diabetic cerebral ischemia/reperfusion injury. CTRP6 overexpression enhanced the sirtuin-1 signaling pathway in diabetic brains after ischemia/reperfusion injury. To investigate the mechanism underlying these effects, we examined mice with depletion of brain tissue-specific sirtuin-1. CTRP6-like protection was achieved by activating the sirtuin-1 signaling pathway. Taken together, these results indicate that CTRP6 likely attenuates cerebral ischemia/reperfusion injury through activation of the sirtuin-1 signaling pathway.

Expression of macrophage inflammatory protein-1αin Kupffer cells following liver ischemia or reperfusion injury in rats

AIM： To explore the expression of macrophage inflammatory protein-1α （MIP-1α） in Kupffer cells （KCs） following liver ischemia/reperfusion injury IRI in rats. METHODS： Forty male SD rats were divided randomly into five groups. A model of partial warm ischemia/ reperfusion injury in the rat liver was established. KCs were isolated and incubated one hour, six hours, 12 h, and 24 h after the reperfusion. Tumor necrosis factor alpha （TNF-α） and interleukin-lbeta （IL-1β） in the supernatants were measured by ELISA. MIP-1α in KCs was detected by immunocytochemical and RT-PCR. RESULTS： No or few MIP-1α protein and mRNA were expressed in the KCs of the control group. Its expression in the IRI group had a significant increase after the reperfusion （P 〈 0.05）, which was contrary to the control group. CONCLUSION： The active behavior of the MIP-1α gene in KCs following liver ischemia/reperfusion injury is assumed to be one of the major causes for the hepatic ischemia/reperfusion injury.

Expression of Macrophage Inflammatory Protein 1α in the Endothelial Cells Exposed to Diamide

In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1α (MIP-1α), the expression of MIP-1α protein in the cells was detected by cell enzyme-linked immunosorbent assay (ELISA) and that of MIP-1α mRNA was determined by cell in situ hybridization and nuclease S1 protection assay after the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activity of MIP-1α was tested by micropore filter method using modified Boyden chambers. Cell ELISA showed that the expression of MIP-1α protein in endothelial cells exposed to 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the control cells, which was statistically significant by analysis of variance. In situ hybridization revealed that the mRNA expression of ECs treated with 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1 3-fold, 3.0-fold and 1.7-fold as much as that in the control group, which had statistical significance ( F =188.93, P <0.01). The mRNA expression in 5 μmol/L dimide treated ECs, measured by nuclease S1 protection assay, was 3.4-fold as much as that in the control group( t =8 70, P <0 05). Chemotactic response(99.50±4.31 μm) to the culture medium conditioned by 5 μmol/L diamide treated ECs , which was stronger than that(66.47±3.25 μm) conditioned by the ECs ( F =404.31, P <0.05), was significantly decreased ( F =192.25, P <0.05) after adding MIP-1α antibody. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce high level of MIP-1α, and might play an important role in atherogenesis by promoting the migration of peripheral blood monocytes into arterial intima.

Effect of monocyte chemoattractant protein-1 on chemotactic gene expression by macrophage cell line U937

Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.

丁酸钠经AMPK/Nrf2/HO-1信号通路调节脂多糖诱导肺泡巨噬细胞极化的作用机制

目的探讨丁酸钠(sodium butyrate,SB)对脂多糖(lipopolysaccharide,LPS)诱导肺泡巨噬细胞极化的影响及其作用机制。方法小鼠肺泡巨噬细胞(MH-S细胞)随机分为对照(Control)组、LPS组、SB组、LPS+SB(LB)组、LPS+SB+腺苷酸活化蛋白激酶(AMPK)抑制剂(Compound C)(LC)组、LPS+SB+核因子E2相关因子2(Nrf2)抑制剂(ML385)(LM)组。通过CCK8检测MH-S细胞活力,筛选出最佳的1000 ng/mL LPS、1 mmol/L SB、10μmol/L Compound C、5μmol/L ML385药物浓度进行后续实验;实时荧光定量(qRT-PCR)检测MH-S细胞白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)、白细胞分化抗原86(CD86)、巨噬细胞甘露糖受体(CD206)、AMPK、Nrf2和血红素加氧酶1(HO-1)的mRNA表达水平;酶联免疫吸附试验(ELISA)检测培养基上清IL-6、TNF-α、IL-1β和IL-10蛋白含量;流式细胞术测定M1和M2型巨噬细胞相关标记物CD86和CD206的表达。各组数据通过单因素方差分析和Tukey法进行检验。结果通过CCK8选取了1000 ng/mL LPS、1 mmol/L SB、10μmol/L Compound C和5μmol/L ML385进行造模和干预。qRT-PCR和ELISA结果一致显示,与LPS组比较,LB组M1型巨噬细胞相关促炎细胞因子IL-6、TNF-α、IL-1β显著降低(均P<0.01),但M2型巨噬细胞相关抑炎细胞因子IL-10显著升高(均P<0.01)。qRT-PCR和流式细胞术结果一致显示,与Control组比较,LPS组CD86水平显著升高(均P<0.01),SB组差异无统计学意义;与LPS组比较,LB组CD86表达水平显著降低(均P<0.01),但M2型巨噬细胞标记物CD206的变化趋势与CD86相反。qRT-PCR结果显示,与LPS组比较,LB组促进AMPK/Nrf2/HO-1的表达(均P<0.05);与LB组比较,LC组降低了AMPK/Nrf2/HO-1的表达(均P<0.05),LM组降低了Nrf2/HO-1的表达(均P<0.05)。流式细胞术结果显示,与LB组比较,LC组和LM组逆转了SB对CD86水平的抑制作用(均P<0.01);M2型巨噬细胞标记物CD206的表达趋势与M1型巨噬细胞标记物CD86相反。结论SB通过激活AMPK/Nrf2/HO-1信号通路,抑制LPS诱导的M1型、促进M2型肺泡巨噬细胞极化,改善了炎症反应。

Reveal more mechanisms of precondition mesenchymal stem cells inhibiting inflammation

Hypoxia can get more ability to inhibit inflammation.But how it impact on survival time of mesenchymal stem cells(MSCs)is confusing and how preconditioned MSCs inhibiting inflammation are partially known.Those issues decided the value of preconditioned MSCs by hypoxia.

Transient Receptor Potential Ankyrin 1 Ion Channel Facilitates Acute Inflammation Induced by Surgical Incision in Mice

Background: Transient receptor potential ankyrin (TRPA) 1 is known as a peripheral initiator of acute inflammation and hyperalgesia. However, its role in the facilitation of innate immune responses followed by resolution of the inflammation triggered by a surgical incision has not been fully investigated. Therefore, we evaluated the mechanism by which TRPA1 regulates the inflammatory responses mainly facilitated by neutrophils and macrophages in the early course of wound repair after an incision. Methods: Plantar incision was performed in wild-type and TRPA1-/- mice. The infiltration of polymorphonuclear neutrophils, macrophage phenotype, and induction of inflammatory mediators were assessed for 7 days postoperatively. Results: TRPA1-/-?mice exhibited decreased infiltration of polymorphonuclear neutrophilscompared with wild-type mice on day 1. Consistently, the influx of F4/80+ iNOS+ proinflammatory M1 macrophages to incised sites was markedly decreased on day 2. Similarly, F4/80+ CD206+M2 macrophages, which regulate the resolution of inflammation and promote wound healing in the later phase of acute inflammation, were significantly decreased in TRPA1-/- compared with those in wild-type mice on day 7. In addition, the induction of heme oxygenase-1, which promotes wound healing by switching phenotype of macrophages, was decreased in the early phase of acute inflammation, whereas the expression of proinflammatory mediators such as tumor necrosis factor and cyclooxygenase-2, and 15-lipoxygenase, which are involved in the resolution of inflammation was increased in the late phase in TRPA1-/- mice. Conclusions: Early innate immune responses including neutrophil infiltration and macrophage polarization at incised sites were inhibited in TRPA1-/- mice, associated with increased pro-inflammatory mediators in later phase. Peripheral TRPA1 may facilitate the acute inflammatory process, leading to the promotion of macrophage-mediated resolution of inflammation and wound repair after a surgical incision.

Are M1 and M2 macrophages Effectual Players in Pathological Conditions?

Pathologic inflammatory conditions are frequently correlated with dynamic alterations through macrophage activation,with classically activated Ml cells associated with promoting and sustaining inflammation and M2 cells implicated in resolving or smoldering chronic inflammation.Inflammation is a common feature of various chronic diseases,and it has direct involvement in the emergence and progression of these conditions.Macrophages participate in an autoregulatory loop characterizing inflammatory process,as they produce a wide range of biologically active mediators that exert either deleterious or beneficial effects during inflammation.Therefore,balancing the ratio of M1/M2 macrophages can help to ameliorate the inflammatory landscape of pathological conditions.This review will explore the role of macrophage polarization in distant pathological inflammatory conditions,such as cancer,autoimmunity,renal inflammation,stroke,and atherosclerosis,while sharing macrophage-driven pathogenesis.

TAK1对非酒精性脂肪肝小鼠的保护作用及其对巨噬细胞极化的影响

目的:探讨转化生长因子β激活激酶-1(TAK1)在非酒精性脂肪性肝病(NFALD)中的作用及其对巨噬细胞极化的影响。方法:将80只5~6周龄C57BL/6雄性小鼠随机分为对照组、模型组、低剂量组和高剂量组。其中,对照组小鼠给予正常饮食,模型组小鼠给予高脂饮食,低剂量和高剂量组小鼠在高脂饮食的基础上,每周腹腔注射1 mg/kg和5 mg/kg的TAK1抑制剂5Z-7-oxozeaenol。持续饲养12周后,HE染色评估肝脏形态。测量小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白(LDL-C)含量。流式细胞术检测小鼠肝脏组织巨噬细胞极化情况。RT-PCR法检测小鼠肝脏组织CD11c、CD206、IL-1β和IL-10 mRNA表达。ELISA检测小鼠肝脏组织IL-1β、IL-10、TNF-α和TGF-β1含量。结果:HE染色结果显示,对照组小鼠肝细胞排列整齐,无脂肪变性和炎症细胞浸润;模型组小鼠肝细胞内可见明显脂肪变性,NAFLD活动度积分显著升高(P<0.05);低剂量组和高剂量组小鼠脂肪变性和炎症细胞显著减少,NAFLD活动度积分明显降低(P<0.05)。与对照组相比,模型组小鼠血清ALT、AST、ALP、TC、TG和LDL-C含量均显著升高,HDL-C含量显著降低(P<0.05);与模型组相比,低剂量组和高剂量小鼠血清ALT、AST、ALP、TC、TG和LDL-C含量显著降低,HDL-C含量显著升高(P<0.05)。与对照组相比,模型组小鼠肝脏组织M1/M2、M1型巨噬细胞标志物CD11c和IL-1βmRNA均明显升高,M2型巨噬细胞标志物CD206和IL-10 mRNA表达降低(P<0.05);与模型组相比,低剂量和高剂量组小鼠肝脏组织M1/M2、CD11c和IL-1βmRNA表达均显著降低,CD206和IL-10 mRNA表达升高(P<0.05)。与对照组相比,模型组小鼠肝脏组织IL-1β和TNF-α表达明显升高,IL-10和TGF-β1表达明显降低(P<0.05);与模型组相比,低剂量和高剂量组小鼠肝脏组织IL-1β和TNF-α表达明显降低,IL-10和TGF-β1表达明显升高(P<0.05)。结论:TAK1抑制剂通过调控巨噬细胞极化,抑制肝脏炎症,发挥肝脏保护作用。

Anti-inflammatory pathways and alcoholic liver disease: Role of an adiponectin/interleukin-10/heme oxygenase-1 pathway

The development of alcoholic liver disease (ALD) is a complex process involving both the parenchymal and non-parenchymal cells in the liver. Enhanced inflammation in the liver during ethanol exposure is an important contributor to injury. Kupffer cells, the resident macrophages in liver, are particularly critical to the onset of ethanol-induced liver injury. Chronic ethanol exposure sensitizes Kupffer cells to activation by lipopolysaccharide via Toll-like receptor 4. This sensitization enhances production of inflammatory mediators, such as tumor necrosis factor-α and reactive oxygen species, that contribute to hepatocyte dysfunction, necrosis, apoptosis, and fibrosis. Impaired resolution of the inflammatory process probably also contributes to ALD. The resolution of inflammation is an active, highly coordinated response that can potentially be manipulated via therapeutic interventions to treat chronic inflammatory diseases. Recent studies have identif ied an adiponectin/interleukin-10/heme oxygenase-1 (HO-1) pathway that is profoundly effective in dampening the enhanced activation of innate immune responses in primary cultures of Kupffer cells, as well as in an in vivo mouse model of chronic ethanol feeding. Importantly, induction of HO-1 also reduces ethanol-induced hepatocellular apoptosis in this in vivo model. Based on these data, we hypothesize that the development of therapeutic agents to regulate HO-1 and its downstream targets could be useful in enhancing the resolution of inflammation during ALD and preventing progression of early stages of liver injury.

Effect of pharmacological intervention on MIP-1α,MIP-1β and MCP-1 expression in patients with psoriasis vulgaris

Objective:To detect the expression level of macrophage inflammatory protein-1(MIP-1)α,MIP-1βand monocyte chemoattractant protein-1(MCP-1)in with psoriasis vulgaris and explore the role in the pathogenesis of psoriasis vulgaris.Methods:The level of MIP-1α,MIP-1βand MCP-1 in peripheral blood from 50 patients with psoriasis vulgaris and 50 normal controls were measured by enzyme linked immunosorbent assay.The correlation with psoriasis area and severity index(PASI)was analyzed.The level of MIP-1α,MIP-1βand MCP-1 was compared between psoriasis vulgaris patients at active stage and resting stage.And the change in MIP-1α,MIP-1βand MCP-1 before and after therapy was also observed.Results:The content of MIP-1α,MIP-1βand MCP-1 in patients with psoriasis vulgaris was(1342.78±210.30),(175.28±28.18)and(266.86±32.75)ng/L,respectively,significantly higher than those in control group(P<0.05).The expression level of MIP-1α,MIP-1βand MCP-1 in peripheral blood of patients with psoriasis vulgaris was positively correlated wilh PASI(P<0.01).After acitretin therapy,expression level of MIP-1α,MIP-1βand MCP-1 in peripheral blood of patients with psoriasis vulgaris was significantly decreased.Conclusions:Chemokine factor MIP-1α,MIP-1βand MCP-1 may be involved in the pathogenesis of psoriasis vulgaris.

Association of NFKB1 gene polymorphism (rs28362491) with levels of inflammatory biomarkers and susceptibility to diabetic nephropathy in Asian Indians

AIM To investigate the association of NFKB1 gene-94 ATTG insertion/deletion(rs28362491) polymorphism with inflammatory markers and risk of diabetic nephropathy in Asian Indians.METHODS A total of 300 subjects were recruited(100 each), normoglycemic,(NG); type 2 diabetes mellitus(T2DM) without any complications(DM) and T2 DM with diabetic nephropathy [DM-chronic renal disease(CRD)]. Analysis was carried out by polymerase chain reaction-restriction fragment length polymorphism and ELISA. Pearson's correlation, analysis of variance and logistic regression wereused for statistical analysis.RESULTS The allelic frequencies of-94 ATTG insertion/deletion were 0.655/0.345(NG), 0.62/0.38(DM) and 0.775/0.225(DM-CRD). The-94 ATTG ins allele was associated with significantly increased levels of urinary monocyte chemoattractant protein-1(u MCP-1); u MCP-1(P = 0.026) and plasma tumor necrosis factor-alpha(TNF-α); TNF-α(P = 0.030) and almost doubled the risk of diabetic nephropathy(OR = 1.91, 95%CI: 1.080-3.386, P = 0.025).CONCLUSION-94 ATTG ins/ins polymorphism might be associated with increased risk of developing nephropathy in Asian Indian subjects with diabetes mellitus.

Effect of glucocorticoid on MIP-1α and NF-кb expressing in the lung of rats undergoing mechanical ventilation with a high tidal volume

BACKGROUND： Ventilator induced lung injury （VILI） is a serious complication in the treatment of mechanical ventilating patients, and it is also the main cause that results in exacerbation or death of patients. In this study, we produced VILI models by using glucocorticoid in rats with high tidal volume mechanical ventilation, and observed the content of macrophage inflammatory protein-1α （MIP-1α） in plasma and bronchoalveolar lavage fluid （BALF） and the expression of MIP-1α mRNA and nuclear factor-kappa B （NF-кB） p65 mRNA in the lung so as to explore the role of glucocorticoid in mechanical ventilation.METHODS： Thirty-two healthy Wistar rats were randomly divided into a control group, a ventilator induced lung injury （VILI） group, a dexamethasone （DEX） group and a budesonide （BUD） group. The content of MIP-1a in plasma and BALF was measured with ELISA and the level of MIP-1α mRNA and NF-кBp65 mRNA expressing in the lung of rats were detected by RT-PCR. The data were expressed as mean±SD and were compared between the groups.RESULTS： The content of MIP-1α in plasma and BALF and the level of MIP-1α mRNA and NF-кBp65 mRNA in the lung in the DEX and BUD groups were signifi cantly lower than those in the VILI group （P〈0.001）. Although the content of MIP-1α in plasma and BALF and the level of MIP-1α mRNA and NF-кBp65 mRNA in the lung in the BUD group were higher than those in the DEX group, there were no signifi cant differences between them （P〉0.05）.CONCLUSIONS： Glucocorticoid could down-regulate the expression of MIP-1α by inhibiting the activity of NF-кB in the lung and may exert preventive and therapeutic effects on VILI to some extent. The effect of local use of glucocorticoid against VILI is similar to that of systemic use, but there is lesser adverse reaction.

Mpeg1在病原清除中的作用及分子机制

巨噬细胞表达基因1(macrophage-expressed gene 1,Mpeg1)编码蛋白是攻膜复合物/穿孔素(membrane attack complex/perforin,MACPF)超家族中的一种穿孔蛋白,广泛存在于动物体内,被认为是现存最古老的MACPF穿孔蛋白。Mpeg1参与宿主先天性免疫,对细菌、病毒和寄生虫均具有杀伤作用。在抗菌免疫中,高表达Mpeg1的细胞可抵御胞内菌、胞外菌、耐酸菌、耐药菌等各种致病菌,这种广谱杀菌活性限制病原菌在宿主体内传播和感染,并间接抑制慢性进行性疾病的发展。在抗寄生虫免疫中,Mpeg1能直接杀伤刺激隐核虫和迈阿密虫等原虫。在抗病毒免疫中,细胞因子诱导表达Mpeg1以抵御病毒感染。近几年的研究表明,Mpeg1还在炎症反应、抗原呈递和一些肿瘤疾病的初步诊断中发挥重要作用。Mpeg1发挥病原清除作用的分子机制为:被诱导表达的Mpeg1通过泛素化在细胞内再分布,最终在溶酶体中与病原体共定位,酸性环境触发Mpeg1在细菌表面打孔,各种抗微生物效应物进入病原菌共同参与病原清除。笔者对Mpeg1编码蛋白的发现、病原清除及其他免疫相关功能及其分子作用机制进行综述,以期为Mpeg1的深入研究和应用提供参考。

TBC1D1 is an energy-responsive polarization regulator of macrophages via governing ROS production in obesity

Energy status is linked to the production of reactive oxygen species(ROS)in macrophages,which is elevated in obesity.However,it is unclear how ROS production is upregulated in macrophages in response to energy overload for mediating the development of obesity.Here,we show that the Rab-GTPase activating protein(Rab GAP)TBC1D1,a substrate of the energy sensor AMP-activated protein kinase(AMPK),is a critical regulator of macrophage ROS production and consequent adipose inflammation for obesity development.TBC1D1 deletion decreases,whereas an energy overload-mimetic non-phosphorylatable TBC1D1^(S231A)Amutation increases,ROS production and M1-like polarization in macrophages.Mechanistically,TBC1D1 and its downstream target Rab8a form an energy-responsive complex with NOX2 for ROS generation.Transplantation of TBC1D1^(S231A)bone marrow aggravates diet-induced obesity whereas treatment with an ultra-stable Tt SOD for removal of ROS selectively in macrophages alleviates both TBC1D1~(S231A)mutation-and diet-induced obesity.Our findings therefore have implications for drug discovery to combat obesity.

The clinical significance of dynamic changes of MIP-1α, MIP-1βand MCP-1 levels in patients with acute pancreatitis

Objective:To observe dynamic changes of MIP-1α, MIP-1β and MCP-1 in Acute pancreatitis patients, and to evaluate the value of MIP-1α, MIP-1β and MCP-1 in acute pancreatitis severity assessment.Methods: A total of 112 cases of acute pancreatitis were divided into mild pancreatitis (MAP group, 44 cases), moderate severe acute pancreatitis group (MSAP group, 36 cases) and severe acute pancreatitis group (SAP group, 32 cases). Serum MIP-1α, MIP-1βand MCP-1 levels were detected by enzyme-linked immunosorbent assay in patients at 1st, 4th day, 7th days.Results:(1) In 1st day, Serum concentrations of MIP-1α, MIP-1β and MCP-1 were significantly increased in three Groups, There were significant differences between the control group, MAP, MSAP and SAP group. (2) Serum concentrations of MIP-1α, MIP-1β and MCP-1 reached the peak level on 4th day in MAP group and were significantly higher than the level of those on 1st day. In the MSAP and SAP group, serum concentration of MIP-1α, MIP-1β and MCP-1 reached the peak level on 1st day, and significantly higher than the level of those on the 4th day and 7th day. There was a significant difference between MSAP group and MAP group, SAP group and MSAP group in the serum concentration of MIP-1α, MIP-1β and MCP-1 at each monitoring time.Conclusions:MIP-1α, MIP-1β and MCP-1 levels have the rule of dynamic change in Patients with acute pancreatitis, which are useful in evaluating the severity of the patients with acute pancreatitis.

Effect of short- and long-term immunization of recombinant disorganized muscle protein-1(rDIM-1) against human filarial parasite Brugia malayi in rodents

Objective: To evaluate the effect of short-term and long-term immunization of recombinant disorganized muscle protein-1(r DIM-1) in rodents against human filarial parasite Brugia malayi.Methods: Recombinant Brugia malayi DIM-1(rDIM-1 bm) protein was cloned, expressed and purified using a Ni-NTA affinity column. Mastomys coucha were immunized with rDIM-1 bm in three immunization schedules: short-term(3-dose of rDIM-1 bm), and long-term(booster doses till 3-and 6-week) and subsequently challenged with infective third-stage larvae of filarial parasite Brugia malayi(L3). Microfilaraemia was monitored in L3 exposed groups on day 90 post larval inoculation(p.l.i.) and continued till day 205 p.l.i. On day 205 p.l.i. all the infected animals were killed and total worm burden was estimated. Cellular proliferative response, macrophage activity, nitric oxide(NO) release, specific IgG and its subtypes, IgE, IgA and Th1(IFN-γ, TNF-ααand IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release were determined. Results: Of the 3 different immunization schedules, shortterm immunization(3-dose schedule) showed better reduction in microfilarial burden(36%-63%) in the peripheral circulation, adult worm load(52%), whereas long-term immunization(3-and 6-week schedule) exerted less effect on peripheral microfilariae count(9%-58%), and adult worm burden(9%-12.5%). Short-term immunization resulted in upregulation of cellular proliferation, macrophages activity, NO release, specific IgG, IgG1, IgG2 a, Ig G2 b, IgE and IgA levels and both Th1(IFN-γ, TNF-α and IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release whereas long-term immunization(3-and 6-week schedule) exerted less effect on parasite burden and showed mixed immunological responses. None of the rDIM-1 bm administration schedules induced any pathology in lymphoid tissues, or alteration in mast cell number and granularity. Conclusions: The short-term immunization with rDIM-1 bm(3-dose schedule) induces robust immune responses and protects the host from filarial parasite infection.