ABSTRACT
ObjectiveLymphatic epithelial cells (LECs) are important links involved in lymphatic metastasis in the microenvironment of cholangiocarcinoma. This study aims to detect the modulation of inflammatory factors and chemokines secreted by LECs after stimulation of cholangiocarcinoma cells, and observe the effects of highly expressed factors on lymphangiogenesis.MethodsThe culture medium of cholangiocarcinoma (RBE, HCCC9810), LECs stimulated by cholangiocarcinoma cell culture medium (CCM), and normal LECs were prepared. Inflammatory factors and chemokines in the culture medium were detected using protein chip. The experiments are divided into the following groups, including a blank control group, CCM group, CCM coupled with Anti-ENA-78 group, Anti-ENA-78 group, ENA-78 group, ENA-78 coupled with SB2252002, and SB225002 group. The relationship between the content of factor and time was investigated using ELISA, while the relation between target factors and lymphangiogenesis obtained by cell proliferation and tubule formation assay.ResultsWe found ENA-78, IP-10, GCP-2, MCP-2, MCP-3, MIP-3a, HCC-1, and Lymphotactin expression increased in LECs supernatant after CCM stimulation. However, I-TAC, MIP-1d, IL-10, MIG, PDGF-BB, and CXCL16 factors showed down-regulation. The secretion of ENA-78 in CCM was relatively low. By ELISA, we found that the ENA-78 protein in RBE-LECs and HCCC9810-LECs gradually increased over time, and reached the plateau phase at the point of 48h. The lymphatic tube forming ability of LECs cultured in CCM was significantly increased compared with that of the control group, and this ability could be partially weakened by ENA-78 neutralizing antibodies. In the exogenous ENA-78 protein group, the lymphatic tube formation ability was as well significantly increased compared with that in the control group, and this ability could be effectively blocked by the IL-8B inhibitor.ConclusionThe increased secretion ENA-78 of lymphatic epithelial cells induced by cholangiocarcinoma may play a role in promoting lymphangiogenesis through the IL-8B receptor.
ABSTRACT
ObjectiveLymphatic epithelial cells (LECs) are important links involved in lymphatic metastasis in the microenvironment of cholangiocarcinoma. This study aims to detect the modulation of inflammatory factors and chemokines secreted by LECs after stimulation of cholangiocarcinoma cells, and observe the effects of highly expressed factors on lymphangiogenesis.MethodsThe culture medium of cholangiocarcinoma (RBE, HCCC9810), LECs stimulated by cholangiocarcinoma cell culture medium (CCM), and normal LECs were prepared. Inflammatory factors and chemokines in the culture medium were detected using protein chip. The experiments are divided into the following groups, including a blank control group, CCM group, CCM coupled with Anti-ENA-78 group, Anti-ENA-78 group, ENA-78 group, ENA-78 coupled with SB2252002, and SB225002 group. The relationship between the content of factor and time was investigated using ELISA, while the relation between target factors and lymphangiogenesis obtained by cell proliferation and tubule formation assay.ResultsWe found ENA-78, IP-10, GCP-2, MCP-2, MCP-3, MIP-3a, HCC-1, and Lymphotactin expression increased in LECs supernatant after CCM stimulation. However, I-TAC, MIP-1d, IL-10, MIG, PDGF-BB, and CXCL16 factors showed down-regulation. The secretion of ENA-78 in CCM was relatively low. By ELISA, we found that the ENA-78 protein in RBE-LECs and HCCC9810-LECs gradually increased over time, and reached the plateau phase at the point of 48h. The lymphatic tube forming ability of LECs cultured in CCM was significantly increased compared with that of the control group, and this ability could be partially weakened by ENA-78 neutralizing antibodies. In the exogenous ENA-78 protein group, the lymphatic tube formation ability was as well significantly increased compared with that in the control group, and this ability could be effectively blocked by the IL-8B inhibitor.ConclusionThe increased secretion ENA-78 of lymphatic epithelial cells induced by cholangiocarcinoma may play a role in promoting lymphangiogenesis through the IL-8B receptor.
ABSTRACT
Objective To observe the changes of ENA -78 expression levels in patients with endometriosis (EM)before and after treatment,and the role of ENA -78 expression in pain and pelvic inflammation.Methods A total of 160 patients with EMwere selected as the research subjects for I phase,II phase,III phase,IV phase patients.In the same period,40 cases of non EMpatients were selected as control group.Serum ENA -78 expression levels were detected among patients with EM before treatment,3 months after treatment and 6 months after treatment,and the control group before treatment by the method of ELISA .The correlation between ENA -78 expression and pelvic inflammatory adhesion and pain grading among patients with EMwas analyzed.Results There was significant difference in the level of serum ENA -78 expression between the two groups at different time points (P <0.01).The expression level of serum ENA -78 of case group decreased with time (P <0.01),while the expression level of serum ENA -78 of EM patients with different pain grading was different (P <0.05).The expression level of ENA -78 in severe group was higher than that in mild group and moderate group (P <0.05).The degree of pelvic adhesion in EMpatients was positively correlated with the level of ENA -78 expression (rs =0.675,P =0.000).The expression of ENA -78 in serum of EMpatients with different degrees of pelvic inflammation was different (P <0.05 ),and the expression of V -grade group ENA -78 was higher than other <br> groups (P <0.05).The level of ENA -78 expression was positively correlated with the degree of pain in patients with EM (rs =0.601,P =0.000).Conclusion Serum ENA -78 expression levels among EM patients were high,and positively correlated with pelvic inflammatory adhesion and pain grading.Serum ENA -78 decreased after GnRh -a treatment.
ABSTRACT
Objective To observe the expressions of grouth-related oncogen (GRO)α, epithelial neutrophil activating protein-78 (ENA-78) and neutrophil-activating peptide-2 (NAP-2) of rat asthma. And to investigate the role of neutrophil in the pathogenesis of asthma exacerbation. Methods In this experi-ment, the rat model of asthma were randomly divided into two groups on average, including asthma group and control group. Levels of ENA-78 at blood neutrophil were detected by flow cytometry method. The ex-pressions of GROα protein at bronchial wall and NAP-2 protein at blood neutrophil were detected by immuno-histochemieal method. Results Levels of GROα, ENA-78 and NAP-2 proteins in asthma group [0.138 ±0.009(A value), 97.65±13.99(MFI), 0.198±0.016(A value), respectively]were significantly higher than those in control group[0.077±0.010(A value), 50.79±8.66(MFI), 0.079±0.015(A value), re-spectively], all P < 0.01. Conclusion Levels of GROα, ENA-78 and NAP-2 were increased at rat asth-ma. They may be participate in inflammation of asthma exacerbation. Neutrophil may promote inflammatory cells influxing into airway wall via increasing synthesis of CXC chemotactic factors.
ABSTRACT
Endometriosis is a common gynecologic disorder defined by the presence,growth,and invasion of endometrial tissue outside the uterine cavity.Although many hypotheses have been proposed,the etiology of the disease remains an enigma.Recent studies have suggested that factors within the peritoneal fluid contribute to endometriotic implant invasion,neovascularization,and proliferation.Chemokines,especially ENA-78,similar to IL-8,are postulated to play an essential role in the local inflammatory reactions seen in endometriosis.
ABSTRACT
BACKGROUND: Inflammation, where vascular endothelial cells are activated by cytokines, recruits circulating leukocytes such as neutrophils into the tissues. Mononuclear phagocytes as well as tissue cells activated by these stimuli produce these chemokines. In this study, the effects of IL-1 and LPS on the expression of CXC chemokines such as GRO-alpha, IL-8 and ENA-78 in vascular endothelial cells and the neutrophil adnesion effects of ENA-78 and GRO-alpha was investigated. METHODS: Human umbilical vein endothelial cells were cultured and stimulated with various concentrations of IL-1 and LPS. The concentrations of the GRO-alpha, IL-8 and ENA-78 secreted were measured using enzyme-linked immunosorbent assay. The effects of ENA-78 and GRO-alpha on neutrophil adhesion to the endothelial cells were also investigated. RESULTS: The addition of IL-1 and LPS to the vascular endothelial cells induced GRO-alpha, IL-8 and ENA-78 secretion in a time- and dose-dependent manner. The neutrophil adhesion was also increased by induction of ENA-78 and GRO-alpha to the vascular endothelial cells in a dose-dependent manner. CONCLUSION: CXC chemokines such as GRO-alpha, IL-8 and ENA-78 secreted by the vascular endothelial cells play an important role in the acute inflammatory responses by stimulating neutrophil adhesion to the vascular endothelial cells, raising the possibility that the CXC chemokines are one of the targets in the clinical application of acute inflammation.