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.

The Expression of Interleukin-17, Interferon-gamma, and Macrophage Inflammatory Protein-3 Alpha mRNA in Patients with Psoriasis Vulgaris

Summary: To investigate the role of Interleukin-17 (IL-17), Interferon-gamma (IFN-γ), and macrophage inflammatory protein-3 alpha (MIP-3α) in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to semi-quantitatively analyze the mRNA expression of IL-17, IFN-γ, and MIP-3α in 31 psoriatic lesions and 16 normal skin tissues. The results showed that the mRNA of the three cytokines was present in all specimens. And the expression level of IL-17 mRNA in skin lesions was 1.1416±0.0591, which was significantly higher than that in normal controls (0.8788±0.0344, P<0.001). The expression levels of IFN-γ mRNA were 1.1142±0.0561 and 0.9050±0.0263, respectively, with significant difference(P<0.001). And the expression levels of MIP-3α mRNA in psoriatic lesions was 1.1397±0.0521, which was markedly higher than that in normal controls (0.8681±0.0308, P<0.001). These findings indicate that up-regulated expression of IL-17, IFN-γ, and MIP-3α might be involved in the pathogenesis of psoriasis.

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.

Neutrophil peptide 1 accelerates the clearance of degenerative axons during Wallerian degeneration by activating macrophages after peripheral nerve crush injury

Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration.However,the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear.This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury.The functions of RAW 264.7 cells we re elucidated by Cell Counting Kit-8 assay,flow cytometry,migration assays,phagocytosis assays,immunohistochemistry and enzyme-linked immunosorbent assay.Axonal debris phagocytosis was observed using the CUBIC(Clear,Unobstructed Brain/Body Imaging Cocktails and Computational analysis)optical clearing technique during Wallerian degeneration.Macrophage inflammatory factor expression in different polarization states was detected using a protein chip.The results showed that neutrophil peptide 1 promoted the prolife ration,migration and phagocytosis of macrophages,and CD206 expression on the surfa ce of macrophages,indicating M2 polarization.The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention.Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α,-6,-12,and tumor necrosis factor-αin invo and in vitro.Thus,the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration,which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.

Th17/Treg balance and macrophage polarization ratio in lower extremity arteriosclerosis obliterans

Objective:To explore the balance of peripheral blood T helper 17 cells/regulatory T cell(Th17/Treg)ratio and the polarization ratio of M1 and M2 macrophages in lower extremity arteriosclerosis obliterans(ASO).Methods:A rat model of lower extremity ASO was established,and blood samples from patients with lower extremity ASO before and after surgery were obtained.ELISA was used to detect interleukin 6(IL-6),IL-10,and IL-17.Real-time RCR and Western blot analyses were used to detect Foxp3,IL-6,IL-10,and IL-17 expression.Moreover,flow cytometry was applied to detect the Th17/Treg ratio and M1/M2 ratio.Results:Compared with the control group,the iliac artery wall of ASO rats showed significant hyperplasia,and the concentrations of cholesterol and triglyceride were significantly increased(P<0.01),indicating the successful establishment of ASO.Moreover,the levels of IL-6 and IL-17 in ASO rats were pronouncedly increased(P<0.05),while the IL-10 level was significantly decreased(P<0.05).In addition to increased IL-6 and IL-17 levels,the mRNA and protein levels of Foxp3 and IL-10 in ASO rats were significantly decreased compared with the control group.The Th17/Treg and M1/M2 ratios in the ASO group were markedly increased(P<0.05).These alternations were also observed in ASO patients.After endovascular surgery(such as percutaneous transluminal angioplasty and arterial stenting),all these changes were significantly improved(P<0.05).Conclusions:The Th17/Treg and M1/M2 ratios were significantly increased in ASO,and surgery can effectively improve the balance of Th17/Treg,and reduce the ratio of M1/M2,and the expression of inflammatory factors.

Macrophage inflammatory protein-2 as mediator of inflammation in acute liver injury

Macrophage inflammatory protein(MIP)-2 is one of the CXC chemokines and is also known as chemokine CXC ligand(CXCL2). MIP-2 affects neutrophil recruitment and activation through the p38 mitogen-activatedprotein-kinase-dependent signaling pathway, by binding to its specific receptors, CXCR1 and CXCR2. MIP-2 is produced by a variety of cell types, such as macrophages, monocytes, epithelial cells, and hepatocytes, in response to infection or injury. In liver injury, activated Kupffer cells are known as the major source of MIP-2. MIP-2-recruited and activated neutrophils can accelerate liver inflammation by releasing various inflammatory mediators. Here, we give a brief introduction to the basic molecular and cellular sources of MIP-2, and focus on its physiological and pathological functions in acute liver injury induced by concanavalin A, lipopolysaccharides, irradiation, ischemia/reperfusion, alcohol, and hypoxia, and hepatectomy-induced liver regeneration and tumor colorectal metastasis. Further understanding of the regulatory mechanisms of MIP-2 secretion and activation may be helpful to develop MIP-2-targeted therapeutic strategies to prevent liver inflammation.

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.

Induction of macrophage inflammatory cytokines by Ox-LDL is ABCA1 dependent

Objective The current study aimed to evaluate whether the induction of macrophage inflammatory cytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages Objective The current study aimed to evaluate whether the induction of macrophage inflammatory eytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages

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.

Role of monocytes and macrophages in experimental and human acute liver failure

Acute liver failure (ALF) is a devastating clinical syndrome characterised by progressive encephalopathy, coagulopathy, and circulatory dysfunction, which commonly leads to multiorgan failure and death. Central to the pathogenesis of ALF is activation of the immune system with mobilisation of cellular effectors and massive production of cytokines. As key components of the innate immune system, monocytes and macrophages are postulated to play a central role in the initiation, progression and resolution of ALF. ALF in humans follows a rapidly progressive clinical course that poses inherent difficulties in delineating the role of these pivotal immune cells. Therefore, a number of experimental models have been used to study the pathogenesis of ALF. Here we consider the evidence from experimental and human studies of ALF on the role of monocytes and macrophages in acute hepatic injury and the ensuing extrahepatic manifestations, including functional monocyte deactivation and multiple organ failure.

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.

血清COL10A1、TK1、MIP-3α水平与胃癌患者病理特征的关系及其对腹膜转移的诊断价值研究

目的研究血清X型胶原α1链(COL10A1)、胸苷激酶1(TK1)、巨噬细胞炎症蛋白-3α(MIP-3α)水平与胃癌患者病理特征的关系及其对胃癌腹膜转移的诊断价值。方法以2021年1月至2022年12月邢台市人民医院收治的96例胃癌患者作为恶性组,其中有腹膜转移27例,无腹膜转移69例;选取同期收治的104例胃良性病变患者作为良性病变组,选取112例健康体检者作为健康对照组。比较各组血清COL10A1、TK1、MIP-3α水平及不同病理特征、有无腹膜转移胃癌患者血清COL10A1、TK1、MIP-3α水平,采用受试者工作特征(ROC)曲线分析血清COL10A1、TK1、MIP-3α对胃癌患者腹膜转移的诊断价值。结果恶性组血清COL10A1、MIP-3α、TK1水平高于健康对照组、良性病变组,良性病变组血清COL10A1、MIP-3α水平高于健康对照组,差异均有统计学意义(P<0.05)。低/未分化、有脉管浸润、肿瘤临床病理分期(TNM)分期Ⅲ~Ⅳ期胃癌患者血清COL10A1、TK1、MIP-3α水平高于高/中分化、无脉管浸润、TNM分期Ⅰ~Ⅱ期患者(P<0.05)。有腹膜转移胃癌患者血清COL10A1、TK1、MIP-3α水平高于无腹膜转移患者(P<0.05)。血清COL10A1、TK1、MIP-3α单项及3项联合检测诊断胃癌腹膜转移的曲线下面积(AUC)分别为0.722(95%CI:0.621~0.809)、0.749(95%CI:0.651~0.832)、0.736(95%CI:0.637~0.821)、0.853(95%CI:0.766~0.917),3项联合检测诊断的AUC大于3项单独检测(Z=1.990、3.617、2.986,P<0.001)。结论胃癌的发生导致患者血清COL10A1、TK1、MIP-3α水平升高,同时随着胃癌患者病情的进展,血清COL10A1、TK1、MIP-3α水平升高,且3项指标联合检测对胃癌患者腹膜转移具有较好的诊断价值。

血清MCP-1和MIP-1β水平与老年脓毒症患者心肌损伤的相关性

目的探讨血清单核细胞趋化因子蛋白1(MCP-1)和巨噬细胞炎性蛋白-1β(MIP-1β)水平与老年脓毒症患者心肌损伤的相关性。方法前瞻纳入2021年3月~2022年3月医院50例出现心肌损伤的老年脓毒症患者,纳入心肌损伤组,同期50例未出现心肌损伤的老年脓毒症患者,纳入非心肌损伤组,测定并比较两组入院时血清MCP-1和MIP-1β水平,分析血清MCP-1和MIP-1β水平与老年脓毒症患者心肌损伤的关系。结果心肌损伤组血清心肌肌钙蛋白T(cTnT)、B型钠尿肽前体(Pro-BNP)、肌酸激酶同工酶(CK-MB)、MCP-1和MIP-1β表达水平高于非心肌损伤组,差异有统计学意义(均P<0.01);经单项Logistic回归分析后建立多元回归模型行多因素分析,结果显示,血清c TnT、Pro-BNP、CK-MB、MCP-1、MIP-1β表达与老年脓毒症患者心肌损伤有关,可能是老年脓毒症患者心肌损伤的预测因素(OR>1,P<0.05);绘制ROC曲线发现,血清MCP-1、MIP-1β表达单独及联合预测老年脓毒症患者心肌损伤的AUC均>0.850,均有一定预测价值。结论血清MCP-1、MIP-1β过表达可能与老年脓毒症患者心肌损伤有关,增加心肌损伤,联合检测老年脓毒症早期血清MCP-1和MIP-1β水平可预测心肌损伤风险。

黄柏碱调节MCP-1/CCR2信号通路对特应性皮炎大鼠炎症反应的影响

目的:探讨黄柏碱对特应性皮炎(Atopic dermatitis,AD)大鼠炎症反应的影响及其作用机制。方法:构建AD大鼠模型,大鼠分为正常组、模型组、黄柏碱低剂量组、黄柏碱高剂量组、黄柏碱+巨噬细胞趋化蛋白-1(Monocyte chemotactic protein-1,MCP-1)组;给药结束后,各组大鼠做皮损评分,记录搔抓次数,HE染色观察皮肤组织病理变化,甲苯胺蓝染色测定肥大细胞数,ELISA试剂盒测定皮肤组织活性氧(Reactiveoxygenspecies,ROS)和血清免疫球蛋白(Immunoglobulins,IgE)、白介素-17(Interleukin,IL-17)、IL-6的含量,Western blot检测皮肤组织MCP-1、CC类趋化因子受体2(CC chemokinereceptor 2,CCR2)蛋白表达。结果:与正常组相比,模型组大鼠脱毛处皮肤发生溃疡、红斑,表皮和棘层增厚且有大量炎性细胞浸润,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平增加(P<0.05);与模型组相比,黄柏碱低、高剂量组皮肤组织红斑、溃疡情况、炎性细胞浸润减少,未见脱屑,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平依次降低(P<0.05);与黄柏碱高剂量组相比,黄柏碱+MCP-1组皮肤红斑、溃疡和炎性细胞浸润加重,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平增加(P<0.05)。结论:黄柏碱可能通过抑制MCP-1/CCR2信号通路降低AD大鼠炎症反应。

血清人分泌型磷脂酶A2、人可溶性髓系细胞触发受体-1、巨噬细胞炎性蛋白3α在妊娠合并获得性重症肺炎患者中的临床应用价值

目的探讨血清人分泌型磷脂酶A2(sPLA2)、人可溶性髓系细胞触发受体-1(sTREM-1)、巨噬细胞炎性蛋白3α(MIP-3α)在妊娠合并获得性重症肺炎(SCAP)疾病中的临床应用价值。方法选择2019年2月至2023年1月我院收治疗80例妊娠合并SCAP患者作为研究组,同期100例妊娠合并获得性非重症肺炎患者作为对照组,比较两组血清sPLA2、sTREM-1、MIP-3α水平、急性生理学及慢性健康状况评分系统II(APACHE-II)评分及孕妇妊娠结局与新生儿结局,经Spearman分析妊娠合并SCAP患者血清sPLA2、sTREM-1、MIP-3α水平与APACHE-II评分的关系;采用多元Logistic回归分析影响妊娠合并SCAP患者新生儿结局发展的因素。结果研究组血清sPLA2、sTREM-1、MIP-3α水平及APACHE-II评分明显高于对照组,其孕产妇不良妊娠结局发生率以及新生儿出现感染、窒息、败血症、宫内窘迫、新生儿肺炎发生率均高于对照组(P<0.05);经Spearman分析发现妊娠合并SCAP患者sPLA2、sTREM-1、MIP-3α水平与APACHE-II评分存在正相关(P<0.05);多元Logistic回归分析显示高水平sPLA2、sTREM-1、MIP-3α水平及APACHE-II评分升高是妊娠合并SCAP患者新生儿不良结局的危险因素(P<0.05);ROC曲线分析提示MIP-3α、sTREM-1、sPLA2均可预测妊娠合并SCAP患者新生儿不良结局,其中sTREM-1的诊断效能最高(P<0.05)。结论血清sPLA2、sTREM-1、MIP-3α与妊娠合并SCAP患者病情发展密切相关,能有效预测新生儿结局发展,可应用于临床。

Del-1纳米颗粒/丝素水凝胶通过促进炎症消退加速慢性皮肤伤口愈合修复

目的 探讨负载发育内皮基因座-1(developmental endothelial locus-1,Del-1)纳米颗粒的丝素水凝胶对小鼠慢性皮肤伤口愈合的影响。方法 在6~8周BALB/c小鼠背部皮肤用磁铁挤压12 h放松12 h,循环4 d,形成慢性压疮;小鼠分为3组,每组8只,分别给予皮肤创面PBS、丝素水凝胶或Del-1纳米颗粒/丝素水凝胶治疗,拍照并计算创面愈合率,连续治疗9 d, HE、Masson染色检测皮肤组织愈合;通过CD14和TNF-α的免疫荧光检测创面巨噬细胞和炎症因子表达;过氧叔丁醇(tert-butyl hydroperoxide, TBHP)体外刺激小鼠巨噬细胞系RAW 264.7和小鼠血管内皮细胞系C166;在C166细胞中过表达Del-1,结晶紫染色检测巨噬细胞迁移;RT-qPCR检测炎症因子IL-6表达。结果 小鼠实验证实Del-1纳米颗粒/丝素水凝胶组的创面愈合快于丝素水凝胶组和PBS组(P<0.01);Del-1纳米颗粒/丝素水凝胶组创面的TNF-α及CD14的表达低于丝素水凝胶组和PBS组(P<0.01),但胶原沉积和组织修复高于PBS和丝素水凝胶组(P<0.01)。体外实验证实TBHP激活巨噬细胞向内皮细胞迁移,但Del-1过表达组巨噬细胞迁移率显著下降(P<0.01);RT-qPCR显示Del-1抑制了IL-6表达(P<0.01)。结论 Del-1纳米颗粒/丝素水凝胶能够加速小鼠压疮愈合,其机制可能是通过促进炎症消退和组织修复。

巨噬细胞上髓系细胞触发受体-1/2在炎症性肠病中的作用研究进展

炎症性肠病(inflammatory bowel disease,IBD)是以克罗恩病(Crohn’s disease,CD)和溃疡性结肠炎(ulcerative colitis,UC)为代表的慢性肠道炎症性疾病,机制涉及遗传易感性以及环境与微生物群间相互作用削弱肠道屏障导致免疫激活等多种途径。近年来,巨噬细胞上表达的TREMs(triggering receptors expressed on myeloid cells),即髓系细胞触发受体,被发现在固有免疫和适应性免疫中发挥重要作用,并与IBD的发生发展密切相关。本文将着重对TREM-1/2(TREM-1和TREM-2)的结构、配体和作用,其在巨噬细胞上参与IBD与伴发精神障碍的机制研究进行概述,旨在为IBD的预防和治疗提供理论支持。

子宫内膜异位症患者血清VEGFR-1和MIP-3α水平表达对术后复发的预测价值研究

目的探究血清血管内皮生长因子受体1(vascular endothelial growth factor receptor-1,VEGFR-1)和巨噬细胞炎性蛋白-3α(macrophage inflammatory protein-3α,MIP-3α)在子宫内膜异位症(endometriosis,EMs)患者中的表达以及两者联合检测对EMs患者术后复发的预测价值。方法选取2019年4月~2021年6月秦皇岛市妇幼保健院行腹腔镜手术治疗的114例EMs患者作为观察组,同期选择在该院体检的孕龄期女性114例健康体检者作为对照组。采用酶联免疫吸附法(ELISA)测定患者血清中VEGFR-1和MIP-3α水平;根据术后二年复发情况,将其分为复发组(n=78)和未复发组(n=36)。采用多因素Logistic回归分析EMs患者术后复发的影响因素;采用受试者工作特征(receiver operating characteristic,ROC)曲线分析血清VEGFR-1与MIP-3α联合检测对EMs患者术后复发的预测价值。结果与对照组相比,观察组VEGFR-1(116.25±48.57pg/ml vs 92.43±25.37pg/ml)及MIP-3α(19.25±5.24pg/ml vs 13.67±4.28pg/ml)水平升高,差异具有统计学意义(t=4.641,8.806,均P<0.05)。轻度、中度、重度EMs患者VEGFR-1水平(104.22±5.78pg/ml,118.60±6.56pg/ml,138.55±7.85pg/ml)和MIP-3α水平(15.37±1.15pg/ml,19.28±2.12pg/ml,25.42±2.56pg/ml)依次升高,差异具有统计学意义(F=147.757,133.654,均P<0.001)。复发组中后穹隆存在触痛结节及r-AFS分期(Ⅲ~Ⅳ期)占比显著大于未复发组(χ^(2)=15.139,10.310,均P<0.05);复发组术后用药6个月及以上占比显著低于未复发组(χ^(2)=15.016,P<0.001),差异具有统计学意义。多因素Logistic回归分析显示,血清VEGFR-1,MIP-3α,后穹隆存在触痛结节及r-AFS分期为EMs术后复发的危险因素(均P<0.05),术后用药6个月及以上为保护因素(P<0.05)。ROC曲线显示,血清VEGFR-1与MIP-3α联合预测EMs术后复发的曲线下面积(area under the curve,AUC)最大(0.929),其敏感度和特异度分别为85.90%和86.11%。结论VEGFR-1及MIP-3α在EMs患者血清中表达升高,且二者联合检测在预测EMs术后复发的效能更佳。

大鼠脊髓损伤后单核细胞趋化蛋白-1、巨噬细胞炎症蛋白-2α基因表达的变化

目的 :探讨大鼠脊髓损伤后单核细胞趋化蛋白 1(MCP 1)、巨噬细胞炎症蛋白 1α (MIP 1α)mRNA表达的变化规律。方法 :SD大鼠 42只 ,随机分为 7组 ,采用改良Allen’s脊髓损伤打击模型 ,以逆转录 -聚合酶链反应 (RT PCR)法测定伤段脊髓MCP 1、MIP 1αmRNA表达情况。结果 :正常脊髓组织内存在MCP 1、MIP 1αmR NA的表达 ,脊髓损伤后MCP 1、MIP 1αmRNA表达逐渐增强 ,MCP 1在伤后 2 4h达到高峰 ,MIP 1α伤后 6h达到高峰。结论 :MCP 1、MIP 1α存在于正常的脊髓组织内 ,脊髓损伤后MCP 1、MIP 1α表达迅速增强 ,提示参与了继发性脊髓损伤过程 ,并可能是损伤因素。

高同型半胱氨酸血症大鼠主动脉平滑肌细胞中巨噬细胞炎性蛋白1α和C-C型趋化因子受体1的表达

目的通过检测高同型半胱氨酸血症大鼠胸主动脉平滑肌细胞中巨噬细胞炎性蛋白1α(MIP-1α)和C-C型趋化因子受体1(CCR1)蛋白的表达,探讨高同型半胱氨酸血症引起动脉粥样硬化(AS)的机制。方法8周龄SD雄性大鼠16只随机分为对照组和高蛋氨酸组,每组8只。对照组给予普通饲料喂养,高蛋氨酸组在普通饲料喂养的基础上加质量分数2%的蛋氨酸。喂养8周后处死大鼠,心脏采血,取胸主动脉。肉眼及光镜下观察血管形态学改变,采用高效液相色谱法测定血浆同型半胱氨酸(Hcy)浓度,S-P免疫组织化学染色法检测MIP-1α和CCR1的表达。结果(1)血浆Hcy浓度:高蛋氨酸组大鼠血浆Hcy浓度(45.74±9.65)mmol.L-1显著高于对照组(7.38±2.28)mmol.L-1(P<0.01)。(2)形态学改变:肉眼观可见对照组大鼠胸主动脉内膜光滑,高蛋氨酸组胸主动脉内膜粗糙、隆起;光镜下可见对照组大鼠主动脉内膜内皮细胞排列整齐,高蛋氨酸组内膜细胞排列紊乱,脂质沉积。(3)免疫组织化学染色结果:MIP-1α和CCR1蛋白的阳性表达表现为细胞质呈棕褐色。MIP-1α和CCR1蛋白在高蛋氨酸组的阳性表达强于对照组,二者在高蛋氨酸组的阳性表达平均灰度值明显低于对照组(P<0.01)。结论高同型半胱氨酸血症可诱导大鼠胸主动脉平滑肌细胞MIP-1α及其受体CCR1的表达,引起血管壁的炎症,导致AS形成。