Effect of Shufeng Tongluo Prescription on Eotaxin and CCR3 Protein Expression and ERK Phosphorylation Level of Asthma Mice / 中国实验方剂学杂志

ObjectiveTo preliminarily explore the mechanism of Shufeng Tongluo prescription （SFTLP） in inhibiting airway inflammation in asthma mice by affecting the expression levels of eotaxin in the serum， CC type chemokine receptor 3 （CCR3）， and extracellular signal-regulated kinase （ERK） phosphorylation in lung tissues. MethodSeventy C57BL/6 mice were randomly divided into a blank group， a model group， low-， medium-， and high-dose SFTLP groups （7.75， 15.5， 30 g·kg-1）， a pertussis toxin （PTX） group， a CCR3 inhibitor （SB328437） group， a phosphoinositide 3-kinase （PI3K） inhibitor （LY294002） group， a p38 protein kinase antagonist inhibitor （SB203580） group， and an ERK inhibitor （PD98059） group. The asthma model was induced in mice by intraperitoneal injection of ovalbumin （OVA） and aluminum hydroxide ［Al（OH）3］ combined with OVA atomization （0.2 mL for all）. After modeling， hematoxylin-eosin staining （HE staining） was used to observe the inflammatory infiltration of lung tissues in mice. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of eotaxin ［CC chemokine ligand （CCL） 11 and CCL24） in each group. Western blot was used to detect the levels of ERK phosphorylation and CCR3 in lung tissues. ResultCompared with the blank group， the model group showed obvious bronchial constriction， lumen stenosis， damaged alveolar structure， massive inflammatory cell infiltration in lung tissues， mucous plug in the bronchus， edema in the submucosal tissues of the trachea， increased folds， increased serum levels of CCL11 and CCL24 （P<0.01）， and increased expression of CCR3 protein in lung tissues （P<0.05）. The ERK levels in lung tissues of the model group and the PTX group increased （P<0.05）. The level of p-ERK in lung tissues of the model group and the low-dose SFTLP group increased （P<0.05）. As revealed by pathological results， compared with the model group， the high-dose SFTLP group showed relieved lung lesions. The high-dose SFTLP group and the SB328437 group showed reduced CCL11 content （P<0.05）. The low- and high-dose SFTLP group， the PTX group， the SB203580 group， the PD98059 group， and the SB328437 group showed decreased CCR3 protein expression in lung tissues （P<0.05）. The high-dose SFTLP group and the PD98059 group showed reduced p-ERK level （P<0.05）. The PD98059 group showed reduced ERK level （P<0.05）. ConclusionSFTLP can inhibit airway inflammation in asthma， and the mechanism may be related to the inhibition of eosinophil activation by down-regulating CCR3 and CCL11 expression and ERK phosphorylation.

The postoperative cognitive dysfunction induced by central inflammation with possible involvement of the gut-brain axis

Abstract Background: Postoperative cognitive dysfunction is widely recognized as severe postoperative central nervous dysfunction and has a significant impact on the 'patient's physical and mental health. Methods: Postoperative models of tibial fracture in aged rats were established, including the control group, model group, CCL11 protein injection group, and saline injection group. Morris water maze test was used to detect the behavioral characteristics of rats. Enzyme-Linked Immunosorbent Assay was used or determine the content of CCL11 and CXCL10. Immunofluorescence staining was used to detect the distribution of CD14+CD163+macro-phages in colon tissues and CD11b+CCR3+microglia cells in hippocampal tissues. Western blot analyzed NOX1 and STAT3 expression in hippocampus tissues. Results: Water maze test results confirmed severe cognitive impairment in CCL11 rats. The content of CCL11 and CXCL10 in the CCL11 group was much higher than that of the model group. The distribution of macrophage and microglia cells in the CCL11 model group was greater than that in the model group and the saline group. The expression of NOX1 and STAT3 in the CCL11 group was higher compared with the model group. Conclusion: Abnormal macrophage function and excessive CCL11 secretion were observed in the rats with lower limb fractures after surgery. Postoperative central inflammation in rats with lower limb fracture induced postoperative cognitive dysfunction through the gut-brain axis molecular mechanism.

Aleitamento materno e quimiocinas: diferenças entre esquizofrenia e controles / Breastfeeding and chemokines: differences in schizophrenia and controls

Introdução: Avaliar a associação entre níveis plasmáticos da quimiocina CCL11, coeficiente de inteligência e prática da amamentação em homens com esquizofrenia em condições psiquiátricas estáveis sob acompanhamento ambulatorial em um serviço de saúde pública. Métodos: Foi realizado estudo caso-controle com 60 indivíduos: 30 pacientes com esquizofrenia e 30 controles saudáveis, dos quais 15 de cada grupo foram expostos ao aleitamento materno e 15 não foram. Foi aplicado questionário abordando questões socioeconômicas, história ao nascer, dados clínicos e alimentação ao nascer. Foi dosada a quimiocina CCL11 e aplicados testes psicológicos para avaliar quociente de inteligência, funcionalidade, sintomas psiquiátricos, curso da doença e diagnóstico. Para os controles, foi utilizada uma escala para descartar doença psiquiátrica. Resultados: A quimiocina CCL11 apresentou valores significativamente mais altos (> 0,5) em pacientes com esquizofrenia quando comparados aos controles. No grupo de amamentados, os esquizofrênicos apresentaram valores significativamente mais altos a nível intermediário (entre 0.106 e 0.5). Não houve correlação da CCL11 com o número de hospitalizações, idade, tempo de diagnóstico e escolaridade. Não foi evidenciada correlação entre tempo de aleitamento materno em relação aos fatores do Brief Psychiatric Rating Scale. Houve uma tendência de correlação entre a idade de início da doença e o aleitamento materno. Foi encontrada correlação positiva do CCL11 com o tempo de aleitamento materno. Ao comparar os pacientes esquizofrênicos que foram aleitados com os que não foram, foi encontrada diferença estatisticamente significativa apenas para o quociente de inteligência. Conclusão: O aleitamento materno está associado a níveis mais baixos de CCL11, escores mais altos de quociente de inteligência e a esquizofrenia. A quimiocina CCL11 é mais alta em quem não amamentou, especialmente nos esquizofrênicos. (AU)

Introduction: To evaluate the association between plasma levels of chemokine CCL11, intelligence quotient, and exposure to breastfeeding in men with schizophrenia under stable psychiatric condition and monitored as outpatients in a public health care unit. Methods: A case-control study of 60 individuals, 30 patients with schizophrenia and 30 healthy controls; in each group, 15 were exposed to breastfeeding and 15 were not. A questionnaire addressing socioeconomic issues, history at birth, clinical data, and feeding at birth was administered. Chemokine CCL11 levels were measured, and psychological tests were applied to assess intelligence quotient, functional status, psychiatric symptoms, disease course, and diagnosis. A scale to rule psychiatric illness was used for the controls. Results: Chemokine CCL11 levels were significantly higher (> 0.5) in patients with schizophrenia than in controls. In the breastfed group, patients with schizophrenia also had significantly higher CCL11 levels, but at an intermediate level (between 0.106 and 0.5). There was no correlation between CCL11 and number of hospitalizations, age, time since diagnosis, or level of education, nor between duration of breastfeeding and the Brief Psychiatric Rating Scale factors. A trend toward a correlation was observed between age at disease onset and breastfeeding. There was a positive correlation between CCL11 and duration of breastfeeding. The comparison of patients with schizophrenia who were breastfed vs those who were not breastfed showed a statistically significant difference only in intelligence quotient. Conclusion: Breastfeeding is associated with lower CCL11 levels, higher intelligence quotient scores, and schizophrenia. Chemokine CCL11 levels are higher in those not exposed to breastfeeding, especially in patients with schizophrenia. (AU)

Effect of cigarette smoke on counts of immunoreactive cells to eotaxin-1 and eosinophils on the nasal mucosa in young patients with perennial allergic rhinitis / Efeito do tabagismo nas contagens de células imunorreativas a eotaxina-1 e eosinófilos na mucosa nasal em pacientes jovens com rinite alérgica perene

Abstract Introduction: In teenagers with perennial allergic rhinitis, exposure to tobacco cigarette smoke increases the count of eosinophils in the nasal mucosa; the recruitment of eosinophils arises from the combined action of a number of cellular and molecular signals, including eotaxin. Objective: To assess the effect of exposure to tobacco cigarette smoke on the count of immunoreactive cells to eotaxin-1 and eosinophils on the nasal mucosa of children and teenagers with perennial allergic rhinitis. Methods: In a cross-sectional study, forty-four patients were evaluated (aged 7-19 years old): 22 with and 22 with no exposure to tobacco cigarette smoke. After replying to 2 validated questionnaires, on Asthma and Allergies in Childhood and on the severity of nasal symptoms, nasal mucosal samples were obtained by scraping the middle one-third of the inferior turbinates. Then counts of immunoreactive cells to eotaxin-1 and eosinophils were assessed by immunohistochemistry. Results: Patients with exposure to tobacco cigarette smoke showed higher cell counts of both eotaxin-1 and eosinophils than patients with no exposure to the smoke, with no correlation between the two variables. However, both counts, of eotaxin-1 and eosinophils, were related to the cotinine/creatinine ratio. Conclusions: Exposure to tobacco cigarette smoke can increase eotaxin-1 and the count of eosinophils in the nasal mucosa of young patients with perennial allergic rhinitis.

Resumo Introdução: Em adolescentes com rinite alérgica perene, a exposição à fumaça do cigarro de tabaco aumenta a contagem de eosinófilos na mucosa nasal. O recrutamento de eosinófilos surge da ação combinada de alguns sinais celulares e moleculares, inclusive a eotaxina. Objetivo: Avaliar o efeito da exposição à fumaça do cigarro de tabaco na contagem de células imunorreativas a eotaxina-1 e eosinófilos na mucosa nasal de crianças e adolescentes com rinite alérgica perene. Método: Em um estudo transversal, 44 pacientes foram avaliados (entre sete e 19 anos): 22 com e 22 sem exposição à fumaça do cigarro de tabaco. Depois de responder a dois questionários validados, sobre asma e alergias na infância e sobre a gravidade dos sintomas nasais, as amostras de mucosa nasal foram obtidas por meio de raspagem do terço médio das conchas inferiores. Em seguida, as contagens de células imunorreativas para eotaxina-1 e eosinófilos foram avaliadas por imuno-histoquímica. Resultados: Os pacientes com exposição à fumaça do cigarro de tabaco apresentaram contagens de células mais elevadas tanto para eotaxina-1 como para eosinófilos em comparação com os pacientes sem exposição à fumaça, sem correlação entre as duas variáveis. No entanto, ambas as contagens, de eotaxina-1 e eosinófilos foram relacionadas com a razão cotinina/creatinina. Conclusões: A exposição à fumaça do cigarro de tabaco pode aumentar a eotaxina-1 e a contagem de eosinófilos na mucosa nasal de pacientes jovens com rinite alérgica perene.

Eosinophils in Colorectal Neoplasms Associated with Expression of CCL11 and CCL24

BACKGROUND: A decrease in the number of tissue eosinophils is known to reflect the malignancy potential of neoplastic lesions and even prognosis. Increased levels of the chemokines CCL11 and CCL24 in serum and tissue are also known to have diagnostic value as serum tumor markers or prognostic factors. The aim of this study was to evaluate the correlation between the degree of tissue eosinophilia and the expression of these chemokines in the glandular and stromal cells of colorectal neoplastic lesions ranging from benign to malignant tumors. METHODS: We counted the number of infiltrating eosinophils in neoplastic lesion tissue and we evaluated the expression of CCL11 and CCL24 in glandular cells and stromal cells by immunohistochemical staining. RESULTS: The results showed that the number of eosinophils decreased significantly and the expression of CCL11 and CCL24 in glandular cells decreased with tumor progression, whereas the stromal expression of CCL11 and CCL24 appeared to increase. CONCLUSIONS: The discrepancy in CCL11 and CCL24 expression between glandular cells and stromal cells might shed light on how colorectal cancer evades the immune system, which would enable further development of immunotherapies that target these chemokines. Further research on eosinophil biology and the expression pattern of chemokines in tumor cells is needed.

Multiple Signaling Pathways Contribute to the Thrombin-induced Secretory Phenotype in Vascular Smooth Muscle Cells

We attempted to investigate molecular mechanisms underlying phenotypic change of vascular smooth muscle cells (VSMCs) by determining signaling molecules involved in chemokine production. Treatment of human aortic smooth muscle cells (HAoSMCs) with thrombin resulted not only in elevated transcription of the (C-C motif) ligand 11 (CCL11) gene but also in enhanced secretion of CCL11 protein. Co-treatment of HAoSMCs with GF109230X, an inhibitor of protein kinase C, or GW5074, an inhibitor of Raf-1 kinase, caused inhibition of ERK1/2 phosphorylation and significantly attenuated expression of CCL11 at transcriptional and protein levels induced by thrombin. Both Akt phosphorylation and CCL11 expression induced by thrombin were attenuated in the presence of pertussis toxin (PTX), an inhibitor of Gi protein-coupled receptor, or LY294002, a PI3K inhibitor. In addition, thrombin-induced production of CCL11 was significantly attenuated by pharmacological inhibition of Akt or MEK which phosphorylates ERK1/2. These results indicate that thrombin is likely to promote expression of CCL11 via PKC/Raf-1/ERK1/2 and PTX-sensitive protease-activated receptors/PI3K/Akt pathways in HAoSMCs. We propose that multiple signaling pathways are involved in change of VSMCs to a secretory phenotype.

Eosinophil Cationic Protein and Chemokines in Nasopharyngeal Secretions of Infants with Respiratory Syncytial Virus (RSV) Bronchiolitis and Non-RSV Bronchiolitis

Bronchiolitis is a risk factor for the development of childhood asthma. Eosinophilic inflammation in airways plays an important role in the pathophysiology of both bronchiolitis and asthma. To investigate this inflammation, we measured the eosinophil cationic protein (ECP), regulated on activation normal T-cell expressed and secreted (RANTES) and eotaxin levels in nasopharyngeal secretions (NPS). Twenty-eight patients with RSV bronchiolitis (RSV group), 11 patients with non-RSV bronchiolitis (non-RSV group) and 7 controls were enrolled in this study. ECP, RANTES, and eotaxin levels were measured by enzyme immunoassays. The ECP level in the NPS of the RSV group was significantly higher than that in the NPS of the non-RSV group and controls. RANTES and eotaxin levels in infants with bronchiolitis were significantly higher than those in the controls, but there was no significant difference between the RSV and non-RSV groups. In conclusion, with regard to eosinophilic airway inflammation, as compared with non-RSV bronchiolitis, RSV bronchiolitis may be more similar to childhood asthma.