Objective To observe the influence of triamcinolone acetonide (TA) on the expression of pigment epitheliumderived factor (PEDF) of human retinal pigment epithelial (RPE) cells. Methods Cultured humanRPE cells (4th－6th generations) were treated with four different concentrations of TA (40, 400, 4times;103 and 4times;104 mu;g/L) for three different periods (12 or 24 or 48 hours), the levels of PEDF protein in the cell culture supernatant and cell lysates were determined by Western blot. After the initial experiment, RPE cells were treated with or without tumor necrosis factor-alpha; (TNF-alpha;, 20 ng/ml） for 24 hours, followed by TA (400 mu;g/L) treatment. The levels of PEDF and phospho-p38 mitogen activated protein kinase(p-p38MAPK) protein expression in cell culture supernatant and cell lysates were measured by Western blot. Results TAtreated RPE cells had higher PEDF expression, and 400 mu;g/L TA group had the highest effect (F＝16.98,P＜0.05). 400 mu;g/L TA treatment for one, six or 24 hours, with or without TNF-alpha; pretreatment, could all promote the PEDF expression and inhibit the p-p38MAPK protein expression (F＝16.87, 10.28; P＜0.01). TNF-alpha; pretreatment alone could inhibit PEDF protein expression and promote p-p38MAPK protein expression (F=16.87, 10.28; P＜0.01). Conclusions TA can up-regulate the expression of PEDF, and downregulate the expression of p-p38MAPK in the cultured human RPE cells.
Objective To investigate the effects of histone modification on the expression of chemokines in alveolar epithelial typeⅡ cells ( AECⅡ) in a rat model of chronic obstructive pulmonary disease ( COPD) . Methods 20 SD rats were randomly assigned to a normal control group and a COPD group. The rat model of COPD was established by cigarette smoking. Lung histological changes were observed by HE staining. AECⅡ cells were isolated and identified by alkaline phosphatase staining and electron microscopic. The mRNA expressions of monocyte chemoattractant protein ( MCP) -1, IL-8, and macrophage inflammatory protein ( MIP) -2αwere detected by real-time quantitative PCR. The expression of histone deacetylase ( HDAC) 2 was measured by western blot. Chromatin immunoprecipitation ( ChIP) was used todetect H3 and H4 acetylation, and H4K9 methylation in the promoter region of chemokine gene. Results Compared with the control group, the mRNA expressions of MCP-1, IL-8, and MIP-2αin the COPD group increased 4. 48,3. 14, and 2. 83 times, respectively. The expression of HDAC2 protein in the COPD group wassignificantly lower than in the control group ( 0. 25 ±0. 15 vs. 0. 66 ±0. 15, P lt; 0. 05) . The expression of HDAC2 had a negative correlation with the gene expressions of IL-8, MCP-1, and MIP-2α( r = - 0. 960,- 0. 914, - 0. 928, respectively, all P lt;0. 05) . The levels of H3 and H4 acetylation were higher, and H4K9 methylation level was lower in the promoter region of chemokine gene in the COPD group compared with the control group ( all P lt; 0. 05) . Conclusions MCP-1, IL-8, and MIP-2α participate and promote the lung inflammatory response in COPD. HDAC2-mediated histone modification may play an important role in COPD inflammation.
ObjectiveTo observe the expression of inflammatory cytokines in diabetic rats received posterior sub-Tenon capsule injection of triamcinolone acetonide (TA) and pan-retinal photocoagulation. MethodsA total of 48 Brown Norway rats received intraperitoneal injection of streptozotocin to establish the diabetic model. Diabetic rats were randomly divided into experimental group (20 rats), control group (20 rats) and blank group (8 rats). 50 μl TA or saline was injected into the posterior sub-Tenon capsule immediately after the photocoagulation in the experimental group and the control group, respectively. The blank group received no treatment. The mRNA and protein expression level of retinal vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and tumor necrosis fator-α (TNF-α) were measured by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) at 1, 3, 7 days after laser photocoagulation. ResultsThe mRNA and protein expression of VEGF, IL-6, TNF-α of the experimental group and control group were significantly higher than the blank group, the difference was statistically significant (P < 0.05). The mRNA and protein expression of VEGF, IL-6 and TNF-α of the experimental group were significantly lower than that of the control group. On day 1 after laser photocoagulation, the mRNA expression of VEGF was not statistically significant in the experimental group and control group (P > 0.05), the mRNA and protein expression of VEGF, IL-6, TNF-α of the two groups were statistically significant in the remaining observing time (P < 0.05). ConclusionPosterior sub-Tenon capsule injection of TA can effectively reduce retinal photocoagulation induced VEGF, IL-6, TNF-α expression.
Objective To observe the effects of stromal cellderived factor 1alpha; (SDF-1alpha;) in secondary neovascular glaucoma (NVG) of proliferative diabetic retinopathy (PDR). Methods The vitreous specimens from 25 PDR patients (31 eyes) were collected with 13 NVG eyes and non-NVG 18 eyes. The concentrations of SDF-1alpha; and vascular endothelial growth factor (VEGF) in those specimens were measured by enzyme-linked immunosorbent assay （ELISA）. Human umbilical vein endothelial cells (HUVEC) were treated by different concentrations of SDF-1alpha;and vascular endothelial growth factor (VEGF) in vitro, and the formation of tube cavity-like structure, length of capillarylike structures and 5prime;-bromo-2prime;-deoxyuridine (BrdU) labeling of treated HUVEC were measured. Results The length of HUVEC tube-like and capillarylike structure formation in 10, 100, 1000 ng/ml SDF-1alpha; and 10 ng/ml VEGF groups were longer than that in the control group, the differences were statistically significant (P＜0.01). The A value of BrdU labeling of 10, 100, 1000 ng/ml SDF-1alpha; and 10 ng/ml VEGF groups were increased than that in the control group, the differences were statistically significant (P＜0.01). The vitreous levels of SDF-1alpha; and VEGF of NVG specimens were higher than those in the non-NVG group, the differences were statistically significant (P＜0.01). Conclusions SDF-1alpha; may promote the migration and proliferation of vascular endothelium cells, and participate in the neovascularization process in NVG patients with PDR.
ObjectiveTo observe the concentration of the inflammatory cytokines in vitreous of severe proliferative diabetic retinopathy (PDR) after intravitreal ranibizumab injection (IVR). MethodsA total of 80 PDR patients (80 eyes) were enrolled in this study. The patients were randomly divided into vitrectomy group (group A) and IVR combined with vitrectomy group (group B), 40 eyes in each group. The differences of sex (χ2=0.05), age (t=0.59), duration of diabetes (t=0.36), HbA1c (t=0.13) and intraocular pressure (F=0.81) between two groups were not significant (P>0.05). The eyes in group B received 0.5 mg (0.05 ml) ranibizumab injection at 7 days before operation. The vitreous samples (0.4 ml) were obtained before operation. The concentration of vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, intercellular adhesion molecule-1 (ICAM-1) and connective tissue growth factor (CTGF) were measured by enzyme-linked immunosorbent assays. ResultsThe concentration of VEGF and ICAM-1 were (10.70±3.60), (224.64±90.32) pg/L in group B and (72.38±23.59), (665.61±203.34) pg/L in group A. The differences of VEGF and ICAM-1 concentration between two groups was significant (t=16.34, 12.53; P<0.001). The concentration of IL-6 and IL-8 were (210.64±80.27), (156.00±57.74) pg/L in group B and (45.78±33.82), (41.07±13.82) pg/L in group A. The differences of IL-6 and IL-8 concentration between two groups was significant (t=11.97, 12.24; P<0.001). There was no difference of CTGF concentration between two groups (t=1.39, P=0.17). The CTGF/VEGF in group B was higher than that in group A (t=14.75, P<0.001). ConclusionsOne week after IVR, the concentration of VEGF and ICAM-1 are decreased, while IL-6 and IL-8 increased. There is no obvious change in CTGF, but CTGF/VEGF is increased.
Diabetic retinopathy (DR) is one of the most common and serious diabetic complications, which is the main cause of vision loss in adults. The specific vascular and neuropathology mechanism of DR is not clear. It has been demonstrated that Inflammatory reaction might be take effects in the development and progression of DR. Monocyte chemoattractant protein-1 (MCP-1), as an important chemokine in the inflammatory response process, promotes chemotactic and activating factors, destroys the blood-retinal barrier, causes retinal vascular disease, and activates microglia, which is related to the severity of the disease. With further research on MCP-1, it is possible to use chemokines and their receptors as target cells to control or slow down the progression of DR by reducing or inhibiting the production of MCP-1 in diabetic patients in the early stages of the disease. This study can provide new ideas and new methods about preventing and treating DR.
Objective To investigate the expression of chemokine receptor CXCR7 and the relation between its expression and clinicopathologic characteristics in papillary thyroid carcinoma. Method The expressions of CXCR7 in 79 cases of papillary thyroid carcinoma and their paracancerous tissues，and 33 cases of benign thyroid lesion tissues were detected by immunohistochemistry. Results The positive expression rates of CXCR7 were 0（0/79），65.8%（52/79），and 30.3%（10/33） in the paracancerous tissues，papillary thyroid carcinoma tissues，and benign thyroid lesions tissues，respectively. The positive expression rate of CXCR7 in the papillary thyroid carcinoma tissues was significantly higher than that in the paracancerous tissues （P＜0.05） or benign thyroid lesion tissues（P＜0.05）. The expression of CXCR7 was correlated with lymph node metastasis （P＜0.05）. Conclusion CXCR7 might take part in tumorigenesis，progression，and lymph node metastasis of papillary thyroid carcinoma.
Objective To explore the effect of dendritic cells (DCs) allergized by K-ras mutant peptide on expressions of chemokines CCL19, CCL22, and cytoskeletal protein fascin-1. Methods DCs were derived from peripheral blood in the presence of granuloceyte/macrophage-colony stimulating factor, interleukin (IL) -4 in vitro. The DCs were collected on day 7 after culture, and were divided into non-K-ras mutant peptide group (addition of RPMI 1604 culture solution 50 μg/ml) and K-ras mutant peptide group (addition of K-ras mutant peptide 50 μg/ml). Phenotype was identified by flow cytometry. The morphological structure was observed by scanning and transmission electron microscopies, respectively. The expressions of IL-12, CCL19, and CCL22 were tested continuously by enzyme-linked immunosorbent assay (ELISA). The expression of cytoskeletal protein fascin-1 was determined by Western blot. Results ①The expressions of CD1a, CD80, and CD86 after loading K-ras mutant peptide were higher than that before loading K-ras mutant peptide (Plt;0.01). ②The DCs with petal-like and branch-like profections after loading were observed under scanning electron microscopy; The DCs with irregular shapes, branch-like or burr-like were showed under transmission electron microscopy. ③The expressions of IL-12, CCL19, and CCL22 in the Kras mutant peptide group were higher than those in the non-K-ras mutant peptide group at different times (6, 12, 24, and 48 h) after loading Kras mutant peptide (Plt;0.01). ④The expression of fascin-1 in the K-ras mutant peptide group was also higher than that in the non-K-ras mutant peptide group (Plt;0.01). Conclusion K-ras mutant peptide can promote DC to mature and improve the expression of chemokines and cytoskeletal protein which will strengthen DC migration.
Objective To investigate the expression of eotaxin-1, eotaxin-2 and eotaxin-3 in ARPE-19 human RPE cells after exposure to light. Methods Cultured human RPE cells (5th~10th generations) were divided into lightinduced group and control group. Cells light-induced group were exposed to the blue light at the intensity of (600plusmn;100) Lux for 12 h to establish the light damaged model. Eotaxin-1, eotaxin-2 and eotaxin-3 mRNA and protein were determined by real time polymerase chain reaction and Western blot at 0, 3, 6, 12, 24 hours after light-induced. Results In light-induced groups, mRNA levels of eotaxin-1 and eotaxin-2 were increased at 0 h (t1=6.05.t2=12.561) and 3 h （t1=2.95.t2=3.67） significantly(P＜0.05), but the mRNA level of eotaxin-3 had not changed （t3=1.57 and 1.00 respectively，P＞0.05） at that time. At 6 h （t1=4.73，t2=18.64，t3=28.48）, 12 h （t1=3.11，t2=20.62，t3=18.50）, 24 h （t1=8.25，t2=38.27，t3=18.60）, mRNA levels of eotaxin-1, 2, 3 were increased significantly （P＜0.05）. Except for the eotaxin-3 protein had not changed at 3 h （t3=1.28，P＞0.05）, protein expression of eotaxin-1, 2, 3 were increased significantly （P＜0.05） at 0 h （t1=4.85，t2=5.45，t3=6..21）, 3 h （t1=5.64，t2=4.55）, 6 h （t1=31.60，t2=6.63，t3=7.15）, 12 h （t1=14.09，t2=18.22，t3=15.76）, 24 h （t1=6.96，t2=10.47，t3=12.85）. Conclusion Eotaxin-1, eotaxin-2 and eotaxin-3 expression were increased after Light-damage, corresponding to the time after light exposure. Eotaxin-3 was the most prominent isoform.
ObjectiveTo investigate the expressions and significance of chemokines factor receptors 4 (CXCR4) and chemokines factor receptors 7 (CXCR7) in gastric cancer tissues. MethodsSixty-five patients with gastric cancer who treated in our hospital from January 2011 to June 2013 were retrospectively collected as gastric cancer group, and 20 patients with gastric ulcer were retrospectively collected as control group at the same time. The expressions of CXCR4 and CXCR7 in gastric cancer tissues and normal gastric tissues were measured by immunohistochemistry, and then the relation-ship among expressions of CXCR4/CXCR7 in gastric cancer tissues and clinicopathological features of patients with gastric cancer was explored, as well as its effect on survival. ResultsPositive expression rates of CXCR4 and CXCR7 were identi-fied in 80.00% (52/65) and 84.62% (55/65) of the gastric cancer group, and 5.00% (1/20) and 10.00% (2/20) in control group respectively, and the positive expression rates of CXCR4 and CXCR7 in gastric cancer group were significantly higher than those of control group respectively (χ2=36.65, P<0.01; χ2=38.55, P<0.01). The positive expression rate of CXCR4 in gastric cancer tissues was related with degree of differentiation, T staging, and TNM staging (P<0.05), positive expression rate of CXCR4 in patients with poor differentiation, T3-4 staging, and TNM Ⅲ-Ⅳ staging were higher than corresponding patients with moderate/high degree of differentiation, T1-2 staging, and TNM Ⅰ-Ⅱ staging. The positive expression rate of CXCR7 in gastric cancer tissues was related with degree of differentiation, T staging, and N staging (P<0.05), positive expression rate of CXCR7 in patients with poor differentiation, T3-4 staging, and N1-3 staging were higher than corrsponding patients with moderate/high degree of differentiation, T1-2 staging, and N0 staging. The survival situation was worse in patients with positive expression of CXCR4 and CXCR7 than corresponding patients with negative expression (P=0.01, P=0.01) respectively. ConclusionsCXCR4 and CXCR7 are related to gastric cancer genesis and development. Furthermore, the expressions of CXCR4 and CXCR7 could be used as markers to predict prognosis of gastric cancer. The regulation of CXCR4/chemokine ligand 12 (CXCL12) axis and CXCR7/CXCL12 axis may provide a new targeted therapy for patients with gastric cancer.