Inhibition of CXCR2 alleviates the development of abdominal aortic aneurysm in Apo E-/- mice

ABSTRACT Purpose To investigate the relationship between atherosclerotic abdominal aortic aneurysm (AAA) and CXC chemokine receptor type 2 (CXCR2). Methods Mouse AAA model was established by embedding angiotensin-II pump (1000 ng/kg/min) in ApoE-/- mice. Mice were received SB225002, a selective CXCR2 antagonist, for treatment. Blood pressure was recorded, and CXCR2+ macrophages were examined by flow cytometry analysis. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining was performed to detect cell apoptosis of abdominal aortic aneurysms. Macrophages were isolated from ApoE-/- mice and treated with Ang II and/or SB225002. Dihydroethidium staining was carried out to determine reactive oxygen species (ROS) activity. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the production of IL-1β and TNF-α. The corresponding gene expressions were measured using real-time polymerase chain reaction (PCR), western blot, and immunohistochemistry staining. Results We found that Ang II activated the expression of CXCR2 in monocytes during the formation of AAA. Inhibition of CXCR2 significantly reduced the size of AAA, attenuated inflammation and phenotypic changes in blood vessels. Ang II-induced macrophages exhibited elevated ROS activity, and elevated levels of 1β and TNF-α, which were then partly abolished by SB225002. Conclusions CXCR2 plays an important role in AAA, suggesting that inhibiting CXCR2 may be a new treatment for AAA.

A comprehensive analysis of the angiogenesis-related genes in glioblastoma multiforme vs. brain lower grade glioma / Uma análise abrangente dos genes relacionados à angiogênese no glioblastoma multiforme vs. glioma cerebral de baixo grau

Abstract Brain tumors are one of the most common causes of cancer-related deaths around the world. Angiogenesis is critical in high-grade malignant gliomas, such as glioblastoma multiforme. Objective: The aim of this study is to comparatively analyze the angiogenesis-related genes, namely VEGFA, VEGFB, KDR, CXCL8, CXCR1 and CXCR2 in LGG vs. GBM to identify molecular distinctions using datasets available on The Cancer Genome Atlas (TCGA). Methods: DNA sequencing and mRNA expression data for 514 brain lower grade glioma (LGG) and 592 glioblastoma multiforme (GBM) patients were acquired from The Cancer Genome Atlas (TCGA), and the genetic alterations and expression levels of the selected genes were analyzed. Results: We identified six distinct KDR mutations in the LGG patients and 18 distinct KDR mutations in the GBM patients, including missense and nonsense mutations, frame shift deletion and altered splice region. Furthermore, VEGFA and CXCL8 were significantly overexpressed within GBM patients. Conclusions: VEGFA and CXCL8 are important factors for angiogenesis, which are suggested to have significant roles during tumorigenesis. Our results provide further evidence that VEGFA and CXCL8 could induce angiogenesis and promote LGG to progress into GBM. These findings could be useful in developing novel targeted therapeutics approaches in the future.

Resumo Os tumores cerebrais são uma das causas mais comuns de mortes relacionadas ao câncer em todo o mundo. A angiogênese tem caráter crítico em gliomas malignos de alto grau, como o glioblastoma multiforme. Objetivo: O objetivo deste estudo foi analisar comparativamente os genes relacionados à angiogênese, VEGFA, VEGFB, KDR, CXCL8, CXCR1 e CXCR2 em GBG vs. GBM para identificar distinções moleculares usando conjuntos de dados disponíveis no The Cancer Genome Atlas (TCGA). Métodos: Os dados de sequenciamento de DNA e expressão de mRNA para 514 pacientes com glioma cerebral de baixo grau (GBG) e 592 pacientes com glioblastoma multiforme (GBM) foram adquiridos do TCGA e as alterações genéticas e os níveis de expressão dos genes selecionados foram analisados. Resultados: Identificamos seis mutações KDR distintas nos pacientes GBG e 18 mutações KDR distintas nos pacientes GBM, incluindo mutações missense e nonsense, exclusão de mudança de quadro e região de emenda alterada. Além disso, VEGFA e CXCL8 foram significativamente super-expressos nos pacientes com GBM. Conclusões: VEGFA e CXCL8 são fatores importantes para a angiogênese, os quais parecem ter um papel significativo durante a tumorigênese. Nossos resultados fornecem evidências adicionais de que o VEGFA e o CXCL8 podem induzir a angiogênese e promover o GBG a progredir no GBM. Esses achados podem ser úteis no desenvolvimento de novas abordagens terapêuticas direcionadas no futuro.

Influência dos polimorfismos genéticos (IL10/CXCL8/CXCR2/ NF?B) na susceptibilidade das doenças reumatológicas autoimunes / Influence of genetic polymorphisms (IL-10/CXCL8/CXCR2/ NF?B) on the susceptibility of autoimmune rheumatic diseases

As doenças reumatológicas autoimunes, na maioria das vezes, possuem uma via genética comum para a autoimunidade. Vários genes foram associados com as doenças reumatológicas, para tanto iremos analisar somente alguns genes nos quais há várias evidências da existência de associação com risco ou proteção de doença autoimune. O fator de transcrição nuclear kappa B (NF-kappa B), o qual regula as respostas imunes e inflamatórias, está associado com esclerose sistêmica (ES), artrite reumatoide (AR) e lúpus eritematoso sistêmico (LES), assim como os genes CXCR2 e CXCL8. Já a interleucina 10 (IL-10), que é uma citocina anti-inflamatória, está associada com quase todas as doenças reumatológicas. Neste artigo, revisamos os potenciais papéis desses genes no sistema imunológico e em diversas doenças reumatológicas. Com relação à IL-10, diversos estudos foram realizados, porém em sua maioria contraditórios - alguns encontraram ausência de associação e outros encontraram associação em diferentes polimorfismos do genes. Já em relação ao NF-kappa B, somente foi estudado em AR e LES, e não foram observadas análises significativas relevantes. Os polimorfismos genéticos do gene CXCR2 foram associados com ES, mas não estão associados com AR e LES. Já os polimorfismos genéticos do gene CXCL8 não estão associados com ES, mas estão associados com AR.

The autoimmune rheumatologic disorders mostly have a common genetic path to the autoimmunity. Several genes have been associated with rheumatologic disorders; therefore, we are analyzing just the ones in those containing several evidences of the existence of association with the risk or protection from autoimmune disorder. The nuclear factor kappa beta (NF-kappa B), which regulates the autoimmune and anti-inflammatory responses, is associated with systemic sclerosis (SS), rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), just as the CXCR2 e CXCL8 genes. On the other hand, the interleukin-10 (IL-10), which is an anti-inflammatory cytokine, is associated with almost all rheumatologic disorders. In this article, we are reviewing the potential roles of these genes in the immune system and in several rheumatologic disorders. In relation to IL-10, several studies have been carried out, but most of them are controversial - some detected the absence of association, and others found association in different genetic polymorphisms. Conversely, in relation to NF-kappa B, it was studied just in RA and SLE, and no relevant significant analyses were observed. The genetic polymorphisms of the CXCR2 gene were associated with SS, but not with RA e SLE. On the other side, the genetic polymorphisms of the CXCL8 gene are not associated with SS, but with RA.

Expression and function of CXCR2, CXCR7 of acute leukemic cells in rat

OBJECTIVE@#To investigate the expression and function of chemokine receptor CXCR2 and CXCR7 in the rat with acute leukemia.@*METHODS@#Flow cytometry and RT-PCR were used to detect the CXCR2, CXCR7 expression on the bone marrow cell surface of the acute leukemia group and the control group.@*RESULTS@#The bone marrow cell surface CXCR2, CXCR7 relative fluorescence intensity of the observation group was significantly higher than the control group (P<0.05). The CXCR7 expression of the extramedullary infiltration group was significantly higher than non-extramedullary infiltration group (P<0.05). The CXCR2, CXCR7mRNA median expression level of the observation group was higher than the control group. The CXCR2 expression and CXCR7 expression of the observation group was positively correlated, and the correlation coefficient was 0.782 (P<0.01).@*CONCLUSIONS@#The chemokine receptor CXCR2 and CXCR7 are highly expressed in acute leukemia, which may be associated with the occurrence of leukemia.

Effect of acupuncture on CXCL8 receptors in rats suffering from embryo implantation failure / 华中科技大学学报（医学）（英德文版）

To observe the effect of acupuncture on CXCL8 receptors (CXCR1 and CXCR2) in rat endometrium experiencing embryo implantation failure, 72 pregnant rats were randomly divided into four groups: normal group (N), embryo implantation failure group (M), acupuncture treatment group (A), and progestin treatment group (W). Then the rats in each group were equally randomized into a day-6 (D6) group, a day-8 (D8) group, and a day-10 (D10) group. The rats in group M, group A, and group W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. Meanwhile, "Housanli" and "Sanyinjiao" were selected for acupuncture. From day 1 to the time of death, the rats in group A were fastened up and then acupuncture was administered while the rats in group N and group M were only fixed, and the rats in group W were given progestin. The number of implanted embryos was calculated. The expression of CXCR1 and CXCR2 in rat endometrium was detected by immunohistochemistry, Western blotting and real-time PCR. Compared to group N, the average number of implanted embryos, the protein and mRNA expression of CXCR1 (D6, D8 and D10), and the protein and mRNA expression of CXCR2 (D8 and D10) in rat endometrium were significantly decreased in group M. Compared to group M, there was significant elevation in the average number of implanted embryos, the protein expression (D6, D8 and D10) and mRNA expression (D8) of CXCR1 in rat endometrium of group A, and the protein expression (D8 and D10) and mRNA expression (D8) of CXCR2 in rat endometrium of group W. These findings indicated that acupuncture can increase the number of implanted embryos in rats of embryo implantation failure, which may be relevant with up-regulation the expression of CXCR1 and CXCR2 at maternal-fetal interface of rats with embryo implantation failure.

Interleukin-8-251 A/T and CXCR2 +1208 C/T genes polymorphisms in chronic rhinosinusitis

IL-8 is one of the pro-inflammatory cytokines which can play an essential role in the pathogenesis of chronic rhinosinusitis [CRS] as well as nasal polyposis [NP]. The ability of individuals in producing IL-8 is partially determined by IL-8-251 A/T polymorphism. Hence, the aim of the present study was to investigate the association between IL-8-251 A/T and CXCR2 +1208 C/T genes polymorphisms and susceptibility to CRS and NP. Two hundred and forty five CRS patients and 204 healthy controls were included in this study. CRS patients were categorized by the existence or absence of NP. IL-8 promoter-251 A/T and CXCR2 +1208 C/T gene polymorphisms were genotyped via the allele specific PCR [AS-PCR] method. While no remarkable difference was demonstrated between patients and controls for both CXCR2 +1208 C/T and IL-8 -251 A/T polymorphisms, a significant increase in IL-8-251 AA genotype was detected in CRS patients with NP compared to those without it [29.3% and 16.2%, respectively; P=0.03]. Interestingly, this association got far stronger when only non-asthmatic CRS patients were taken into consideration [P=0.001]. The results of the present study indicate that the inheritance of IL-8-251 Aallele is associated significantly with NP development in CRS patients. Therefore, NP formation might be a result of the exposure to an intense inflammatory environment, which is more likely in genetically susceptible CRS patients

Cytotoxicity of dichloroacetic acid in lymphocyte and expression of chemokine receptor CXCR2 and chemokine receptor CXCR3 mRNA / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the effects of trichloroethylene (TCE) and its by-products (trichloroacetic acid, TCA; dichloroacetic acid, DCA) on the normal human peripheral blood lymphocyte and the role of DCA in dermatitis medicamentosa- like induced by trichloroethylene (DMLT).</p><p><b>METHODS</b>Lymphocyte was isolated from peripheral venous blood, and cytotoxicity of human lymphocytes treated with different concentrations (0.02 approximately 30.00 mmol/L) of DCA was determined at indicated times (2 h and 4 h) based on the MTS assay. Action of DCA on cell viability, membrane integrity was assessed by neutral red uptake (NRU) assay and lactate dehydrogenase (LDH) release test and measurement of superoxide dismutase (SOD) activity. Fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) was employed for detection and quantization of the chemokine receptor CXCR2 and chemokine receptor CXCR3 mRNA in peripheral blood lymphocyte treated with different concentrations of DCA.</p><p><b>RESULTS</b>DCA had a more vital effect on peripheral blood lymphocyte than TCE and TCA. A concentration-dependent release of LDH was observed at 4 h after cells were exposed to different doses of DCA (0.88, 1.75, 3.50 and 7.00 mmol/L) (P < 0.05), and DCA also caused an inhibition of SOD activity in a concentration-dependent manner (P < 0.05). The results of FQ- RT- PCR indicated that CXCR2 and CXCR3 mRNA were all over- expression. At 48 h after the DCA of 0.5 mmol/L and 10.00 mmol/L was used, CXCR2 and CXCR3 mRNA were 10.34, 5.66-fold and 19.43, 8.75-fold of those in the control group (P < 0.01).</p><p><b>CONCLUSION</b>DCA is of a great cytotoxicity and may be one of crucial evocators on DMLT.</p>

The role of CXCR1/2 in shear stress-induced endothelial cell migration / 生物医学工程学杂志

CXCR1 and CXCR2 are important receptors in regulating vascular endothelial cell activities. In order to elucidate the role of CXCR1/2 in shear stress-induced endothelial cell migration, we have investigated the expression levels of CXCR1 and CXCR2 in the endothelial cells exposed to shear stress. In the experiment, anti-IL8RA and anti-IL8RB were used to antagonize CXCR1 and CXCR2. Different shear stresses were generated in a flow chamber; scratch test was carried out to compare endothelial cell migration in the control group and the receptor-antagonized groups. The results indicated that the migration of endothelial cells was restrained effectively after CXCR1 and CXCR2 were antagonized by anti-IL8RA and anti-IL8RB. And anti-IL8RA showed a stronger inhibitive effect than did anti-IL8RB (P<0.05). In the group with both receptor antagonisms, the migration was further inhibited. These results suggest that both CXCR1 and CXCR2 are important factors in mediating the migration of endothelial cells induced by shear stress, and CXCR1 fulfills a more important role.

Investigation of IL-8Rbeta mRNA expression profile in single human neutrophil / 中国应用生理学杂志

<p><b>AIM</b>To validate the abundance of Interleukin 8 receptor beta (IL-8Rbeta) mRNA in single human neutrophil.</p><p><b>METHODS</b>Human neutrophils were isolated and purified from volunteers, total RNA was extracted and a regular RT-PCR aiming at IL-8Rbeta mRNA was performed to ascertain its expression profile in human neutrophils and optimize the reaction conditions for the following single-cell RT-PCR procedures. Subsequently, single neutrophil or the cellular content was harvested to conduct reverse transcription and two-round PCR with the same primer pairs used before. Serial dilution of single neutrophil cDNA pool was carried out at the same time with the exact two-round PCR followed. The specificity of this single-cell RT-PCR procedure was verified by the BamHI restriction endonuclease digestion on the final cDNA products.</p><p><b>RESULTS</b>Regular RT-PCR indicated IL-8Rbeta mRNA expression in human neutrophils. While single-cell RT-PCR was sensitive enough to detect trace IL-8Rbeta mRNA as predicted cDNA product could be amplified from a 10 000 times diluted intracellular specimen from single neutrophil, which indicated an abundant expression of this mRNA in human neutrophil. Moreover, BamHI digestion on the final cDNA product clarified the specificity of this single-cell RT-PCR procedure.</p><p><b>CONCLUSION</b>This simplified semi-quantitative single-cell RT-PCR procedure specifically confirmed that IL-8Rbeta mRNA was highly expressed in human neutrophil, which also provided the possibility of comparing mRNA abundance at single cell level.</p>

Effects of fluid shear stress on the expression of IL-8 receptor mRNA in EA. Hy926 cells / 生物医学工程学杂志

This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 and CXCR2 mRNA in endothelial cells, EA. Hy926 cells. The HUVEC cell lines were cultured in vitro and then exposed to 5.56, 10.02, and 15.27 dyn/cm2 fluid shear stress respectively for 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 1 h, 2 h, 4 h and 8 h. Semi-quantitative reversal transcription-polymerase chain reaction (RT-PCR) was used for detecting IL-8 receptors mRNA expression at different times. The results showed that, under 5. 56 dyn/cm2 shear stress, both the expression of CXCR1 and CXCR2 mRNA increased significantly with time (P<0.05). When exposed to 10. 02 dyn/cm2, the expression of CXCR1 mRNA was down-regulated with time on every occasion. CXCR2 mRNA increased temporally at 30 min, then it decreased gradually with time and finally went on at a constant lower level. When exposed to 15.27 dyn/cm2, both CXCR1 and CXCR2 mRNA expression decreased significantly with time (P<0.01). These data indicate that the expression of CXCR1 and CXCR2 mRNA of endothelial cell is regulated by fluid shear stress.

Immunohistochemical Study on the Expression of Chemokines and Chemokine Receptors in Psoriasis / 대한피부과학회지

BACKGROUND: Psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes. OBJECTIVE: The purpose is to understand the pathogenetic mechanisms of psoriasis by comparing immunoreactivity of various chemokines and chemokine receptors between lesional and non-lesional skin of psoriasis. METHODS:We have performed immunohistochemical studies with mouse anti-human IL-8, mouse anti-human GRO, anti-huamn MCP-1, mouse anti-human RANTES, anti-human CDw 128 IL-8RA/ CXCR1, and anti-human IL-8RB/CXCR2 for lesional and non-lesional skin of ten psoriatic patients. RESULTS: 1.Immunohistochemical reactivity for IL-8 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for GRO-alpha is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 2.Immunohistochemical reactivity for MCP-1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05), and immunohistochemical reactivity for RANTES is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 3.Immunohistochemical reactivity for CXCR1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for CXCR2 is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 4.Immunofluorescent staining reveals positive finding in epidermis of lesional psoriasis, but negative finding in CXCR2. CONCLUSION: These results suggest that psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and that both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes.

Immunohistochemical Study on the Expression of Chemokines and Chemokine Receptors in Psoriasis / 대한피부과학회지

BACKGROUND: Psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes. OBJECTIVE: The purpose is to understand the pathogenetic mechanisms of psoriasis by comparing immunoreactivity of various chemokines and chemokine receptors between lesional and non-lesional skin of psoriasis. METHODS:We have performed immunohistochemical studies with mouse anti-human IL-8, mouse anti-human GRO, anti-huamn MCP-1, mouse anti-human RANTES, anti-human CDw 128 IL-8RA/ CXCR1, and anti-human IL-8RB/CXCR2 for lesional and non-lesional skin of ten psoriatic patients. RESULTS: 1.Immunohistochemical reactivity for IL-8 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for GRO-alpha is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 2.Immunohistochemical reactivity for MCP-1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05), and immunohistochemical reactivity for RANTES is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 3.Immunohistochemical reactivity for CXCR1 is stronger in lesional epidermis than non-lesional epidermis(p<0.05) and immunohistochemical reactivity for CXCR2 is stronger in lesional epidermis than non-lesional epidermis(p<0.05). 4.Immunofluorescent staining reveals positive finding in epidermis of lesional psoriasis, but negative finding in CXCR2. CONCLUSION: These results suggest that psoriatic keratinocytes express CXC chemokines like IL-8 and GRO-alpha, and CC chemokines like MCP-1 and RANTES, which have a significant role in the accumulation of inflammatory cells in psoriatic skin and that both CXCR1 and CXCR2 receptors are also expressed in psoriatic keratinocytes, which suggests that IL-8 and GRO-alpha could have a role in the characteristic epidermal changes through binding to their receptors in psoriatic keratinocytes.