ObjectiveTo investigate the expression of interleukin-18（IL-18）and signal transducers and activators of transcription 5（STAT5）in retina of 4-24-week-old diabetic rats, and explore the potential molecular mechanisms involved in diabetic retinopathy (DR).MethodsRetinal gene expression profile of healthy and 8-week-old diabetic rats was established with restriction fragment differential displaypolymerase chained reaction (RFDD-PCR), and the differences was analyzed by bioinformatics. IL-18 and STAT5 were filtrated as the candidate genes of DR. The expression of IL-18 and STAT5 in retina of diabetic rats with the age of 4, 8, and 24 weeks was observed by semi-quantitative reverse transcriptase-polymerase chain reaction（RT-PCR）.ResultsThe result of RFDD-PCR showed:expression of IL-18 was higher in healthy retina than that in diabetic one; expression of STAT5 was not found in healthy rats but in diabetic ones. The result of RT-PCR showed：compared with the normal, high expression of IL-18 was found in 4-week diabetic retina, reduced in 8-week one, and decreased to the lowest in 24-week one. The expression of STAT5 was not observed in healthy or 4week diabetic retina, but occurred in 8-week one, and increased in 24-week one. ConclusionThe expression of IL-18 and the activation of STAT5 may relate to the occurrance of DR. The expression of IL-18 doesn′t depend on the activation of STAT5. (Chin J Ocul Fundus Dis, 2005,21:258-260)
ObjectiveTo observe the effect of interleukin (IL) 10 modified endothelial progenitor cells (EPC) in diabetic retinopathy (DR). MethodsEPC cells were collected and cultivated from the bone marrow of rats and identified by immuno-fluorescence staining. EPC cells were infected with lentivirus (LV) of EPC-LV-IL10-GFP (EPC-LV-IL10-GFP group) or EPC-LV-NC-GFP (GFP group). EPC cells without lentivirus infection was the EPC group. Enzyme-linked immuno sorbent assay (ELISA) was used to measure the concentrations of tumor necrosis factor (TNF)-α, IL10, IL8 and vascular endothelial growth factor (VEGF) in the supernatant of these three groups. 168 male Wistar rats were divided into normal control group (28 rats), diabetes mellitus (DM) group (28 rats), DM-blank control group (56 rats) and DM-intervention group (56 rats). DM was introduced in the latter 3 groups by streptozotocin intravenous injection. Three months later, the rats in the DM-blank control group and DM-intervention group were injected with EPC-LV-NC-GFP or EPC-LV-IL10-GFP by tail vein, respectively. Immunohistochemistry was used to observe the GFP expression in rat retinas. The blood-retinal barrier breakdown was detected by Evans blue (EB) dye. The retinal histopathologic changes were observed by transmission electron microscope. The mRNA level of VEGF, matrix metallproteinases-9 (MMP-9), angiopoietin-1 (Ang-1), inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in retina were measured by reverse transcription-polymerase chain reaction (RT-PCR). ResultsELISA showed that the levels of TNF-αand IL8 in the supernatant significantly decreased, while the levels of IL10 and VEGF increased (P < 0.05) in EPC-LV-IL10-GFP group. GFP expressed in the retina of blank control group and intervention group, mainly in the ganglion cell layer, inner nuclear layer and outer plexiform layer. The retinal blood vessel pathological change and EB permeability significantly decreased in intervention group compared with DM group (P < 0.05), and blank control group (P < 0.05). RT-PCR revealed that the mRNA level of VEGF, MMP-9 and Ang-1 significantly increased, and eNOS decreased in DM group compared to the normal control group (P < 0.05). The mRNA level of VEGF and iNOS decreased, eNOS increased while Ang-1 and MMP-9 had not changed in DM-blank control group and DM-intervention group compared with DM group (P < 0.05). ConclusionsIL10 modified EPC can improve the inflammative microenvironment and suppressed the pathogenesis of DR. Furthermore, EPC transplantation can increase the number of EPC and exerted their effect.
Objective To investigate the response of retinal ganglion cells （RGC）in detached and reattached retina in adult rats, and the effect of IL-1beta antibody and IL-1Ra on the loss of RGC. Methods A total of 73 Sprague-Dawley (SD) rats were subretinally injected with healon GV（1.4% hyaluronate）and retrograde labeled with fluorogold (FG), and 10 ng IL-1 Ra and 500 ng IL-1beta antibody were injected into the subretinal space combined with healon GV. The retinal flakes were observed under the fluoroscope and the number of RGC was counted 2 hours, 1, 3, 5, 7, 10, 20, 50, and 90 days after deta chment; 10 days after detachment and 30 days after reattachement; 90 days after detachment and 20 days after reattachement, and 1 and 10 days after injection with IL-1beta antibody and IL-1Ra,respectively. And the control group was only developed an intraocular injection of the same valume of healon GV. Result Two hours after detachment, the RGC loss was found, reached the peak at first day, and decreased gradually. RGC loss was also found in the non-detached area. The reattachment 10 days after detachment (early reattachment) stopped the loss of RGC, and the reattachment 90 days after detachment (late reattachment) promoted the loss, which rested on a certain level. Subretinal space injection of IL-1Ra and IL-1beta antibody decreased the loss of RGCs in the detached retina. Conclusion The RGCs loss were found both in the detached and attached retina. Early reattachment may stop the loss of RGC, and late reattachment may promote the loss. Both IL-1beta antibody and IL-1Ra have neuro protective effect on RGC. (Chin J Ocul Fundus Dis，2004，20：233-236)
Objective To investigate the effect of interleukin-10 （IL-10） gene transfer on expression of CD44, selectin-E, lymphocyte function associated antigen-1 （LFA-1）, vascular cell adhesion molecule-1 （VCAM-1） in mice heart transplantation rejection. Methods Model of mice cervical heterotopic heart transplantation was set up, 96 mice were divided into three groups with random number table, control group： heart transplantation between C57 mice; transplant group： heart from BALB/C mice transplant to C57 mice; IL-10 group： IL-10 was transfected on BALB/C mice isolated heart for 1 hour, then transplanted to C57 mice. The messenger ribonucleic acid （mRNA） level expression of CD44 ,selectin-E ,LFA-1 ,VCAM-1 and IL-10 were measured by reverse transcription-polymerase chain reaction （RT-PCR） at the 5th day after transplantation. Results The mRNA level expression of CD44, selectin-E ,LFA-1 ,VCAM-1 in transplant group were significantly increased than those in control group （P〈0.01）. The mRNA level expression of CD44, selectin-E, LFA-1 ,VCAM-1 in IL-10 group were significantly decreased than those in transplant group （P〈0.01）. Conclusion IL-10 gene transfer is able to decrease the expression of CD44, selectin-E,LFA-1 ,VCAM-1 and suppress the heart transplantation rejection in mice.
【Abstract】Objective To study the influence of early hemofiltration on plasma concentrations of proinflammatory cytokines TNF-α and IL-1β and their transcription levels in severe acute pancreatitis (SAP) pigs. Methods The model of SAP was induced by retrograde injection of artificial bile into pancreatic duct in pigs. Animals were divided randomly into two groups: SAP hemofiltration treatment group (HF group, n=8) and SAP no hemofiltration treatment group (NHF group, n=8). TNF-α and IL-1β plasma concentrations were measured by ELISA. Their transcription levels in the tissues of pancreas, liver and lung were assayed by semi-quantitative reverse transcription polymerase chain reaction. Results After hemofiltration treatment, the plasma concentrations of TNF-α and IL-1β increased gradually but were lower than those of NHF group at the same time spot 〔at 6 h after hemofiltration treatment, (618±276) pg/ml vs (1 375±334) pg/ml and (445±141) pg/ml vs (965±265) pg/ml, P＜0.01〕. At 6 h after hemofiltration treatment, the transcription levels of TNF-α and IL-1β in tissues of pancreas, liver and lung were lower than in NHF group (57.8±8.9 vs 85.7±17.4, 48.0±8.1 vs 78.1±10.2, 46.2±9.6 vs 82.4±10.5; 55.9±9.0 vs 82.2±15.7, 40.6±9.2 vs 60.0±10.6, 35.7±9.8 vs 58.1±9.3, P＜0.01). Conclusion Early hemofiltration can reduce TNF-α and IL-1β plasma concentrations and transcription levels in SAP pigs.
Objective To explore the effects of asiaticoside on the activation of nuclear factor kappa B ( NF-κB) and cytokines expression in RAW264. 7 cells induced by lipopolysaccharide ( LPS) . Methods RAW264. 7 cells were allocated to 5 groups, ie. a blank group, a model group stimulated by LPS at dose of 10 μg/mL, and three asiaticoside treatment groups stimulated by LPS and different doses of asiaticoside simultaneously. The effects of asiaticoside ( 10 - 7 , 10 - 6 , 10 - 5 mol /mL) on the proliferation of cells were examined by MTT assay. The activation of NF-κB was detected and analyzed by the laser scanning confocal microscope( LSCM) ,meanwhile the concentrations of TNF-α, IL-1, and IL-10 in supernatants were quantified by ELISA. Results MTT assay showed that asiaticoside ( 10 - 7 , 10 - 6 ,10 - 5 mol /mL) had no effects on the proliferation of RAW264. 7 cells. Asiaticoside significantly decreased the activation of NF-κB, downregulated the secretion of TNF-αand IL-1, and upregulated IL-10 secretion in a dose dependent manner. According to LSCM, the ratio of NF-κB activation was ( 3. 5 ±1. 5) % , ( 75. 7 ±9. 1) % , ( 66. 8 ±7. 1) % , ( 58. 9 ±9. 0) % , and ( 40. 1 ±8. 8) % in the blank, model, and asiaticoside( 10 - 7 , 10 - 6 , 10 - 5 mol /mL) treatment groups respectively. The contents of TNF-α in supernatants were ( 171. 12 ±35. 42, 1775. 45 ±193. 97,1284. 63 ±162. 13,1035. 22 ±187. 97, 598. 90 ±107. 73) pg/mL respectively and IL-1 were ( 5. 66 ±0. 98,26. 93 ±3. 48,22. 41 ±2. 84, 17. 05 ±1. 70, 10. 64 ±1. 29) ng/mL respectively, while IL-10 were ( 25. 23 ±2. 17,71. 75 ±8. 31, 82. 82 ±6. 00, 98. 70 ±8. 84, 119. 97 ±9. 13) pg/mL respectively. Conclusion The antiinflammation mechanism of asiaticoside may be mediated by downregulating inflammatory factors throughNF-κB signal pathway and keeping the balance between proinflammatory and antiinflammatory system.
Objective To explore the changes and interrelationship of serum interleukin-12 (IL-12) and T lymphocyte subset in patients with primary hepatic carcinoma (PHC). Methods Serum IL-12 level was determined by ELISA in 36 patients with PHC. The peripheral blood T lymphocyte subset was assessed with flow cytometry. The distribution and changes of T lymphocyte subset in the tumor tissue were detected by immunohistochemistry analysis. Results The numbers of the CD＋4 T cell were reduced and of the CD＋8 T cell increased either in peripheral blood or tumor tissue, and showed the trend of the ratio (T4/T8) declined progressively with the aggravation of the state with PHC. IL-12 and T4/T8 had significant interrelationship.Conclusion IL-12 is an important antitumor factor of the patients with PHC. T lymphocyte subset plays a great role in the process of antitumor.
ObjectiveTo investigate the inhibitory effects of L arginine (L arg) on systemic inflammatory response after cardiopulmonary bypass(CPB).MethodsFifty one patients with rheumatic heart disease were randomly divided into two groups: L arg group ( n =25) and control group ( n =26). For L arg group, L arg at 300mg/kg was given during operation. Plasma levels of tumor necrosis factor α(TNF α),interleukin 1β(IL 1β)and interleukin 10(IL 10) were measured by enzyme linked immunosorbent assay technique at baseline(before operation) and at 2,4,8,24 and 48 h after CPB termination.ResultsTNF α,IL 1β and IL 10 levels were increased in both groups after CPB ( P lt;0.05); levels of TNF α, IL 1β returned to normal at 48 h after CPB; In L arg group, TNF α and IL 1β levels were significantly lower than those in control group at 4,8 and 24 h after CPB ( P lt; 0 05). No significant difference were detected in IL 10 between groups( P gt;0.05).ConclusionL arg may decrease plasma levels of TNF α and IL 1β after CPB, it implies L arg may inhibit inflammation induced by CPB.
Interleukin-1 (IL-1), interleukin-2(IL-2) and interleukin-6(IL-6) activities and tumor necrosis factor (TNF) contents in plasma from patients with different sites of cancers as well as controls using bioassay technique were studied. The results showed that the levels of IL-1,IL-2,IL-6 s from patients with different sites of cancer were decreased remarkably in comparision with controls and the contents of TNF from patients with different sites of cancers increased significantly. But the difference between different sites of cancer was not statistically significant. The data suggest that the variations in the contents of TNF and the levels of interleukins may be related to the development of these caner patients.
ObjectiveTo investigate the clinical signification of plasma interleukin-17 (IL-17) 1evel in patients with acute respiratory distress syndrome (ARDS).MethodsForty-five adult ARDS patients and 22 healthy controls were enrolled in this study. The plasma cytokine levels of IL-17, IL-6 and IL-10 were measured by enzyme linked immunosorbent assay. Meanwhile, the baseline data of demographic and clinical tests including oxygenation index, procalcitonin and brain natriuretic peprtide were collected, the acute physiological and chronic health Ⅱ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score were recorded. The main outcome was defined as hospital mortality within 28-day follow-up.ResultsThe plasma concentration of IL-17, IL-6 were higher in the ARDS patients (P<0.05) compared with the controls and the mean levels of IL-17, IL-6 and the APACHEⅡ score and the SOFA score in the non-survivors was higher than those in the survivors (P<0.05). In particular, there was a significant correlation between the plasma levels of IL-17 and IL-6 (P<0.05). Logistic regression and COX multivariate survival analysis suggested that age and SOFA score may be prognostic factors for ARDS.ConclusionsThe plasma concentration of IL-17 is significantly increased in ARDS patients, and its expression is linearly related to the proinflammatory factor IL-6. Both are important inflammatory markers in the acute phase of ARDS and may be important disease severity and prognostic indicators in addition to age and SOFA score.