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.
Objective To investigate the changes of microRNA-150 ( miR-150) in peripheral blood leukocytes in sepsis patients, and their relationship with expression of immune cytokines and sepsis severity. Methods The level of mature miR-150 was quantified by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and normalized to that of control miRNA, U6, in peripheral blood leukocytes of 40 patients with sepsis, 20 patients with systemic inflammatory response syndrome ( SIRS) , and 20 normal individuals. Serum concentrations of tumor necrosis factor alpha (TNF-α) and interleukin-10 (IL-10) were measured by enzyme-linked immunoabsorbent assay in all subjects. The sequential organ failure assessment ( SOFA) score systemwas used to evaluate the severity of sepsis. The relationships between miR-150 and the white blood cell count ( WBC) , TNF-α, IL-10 and SOFA score of the sepsis patients were analyzed. Results MiR-150 was stable for at least 5 days when specimen stored at 4 ℃ and the determination of miR-150 had a broad linear detecting range ( 6. 97-6. 97 ×104 pg/ μL RNA, the lowest detecting limit: 6. 97 pg/μL RNA,r=0.999) .MiR-150 expression in the peripheral blood leukocytes in the sepsis group was significantly lower than that in the healthy control group ( Plt;0.01) , while WBC, IL-10 and IL-10/TNF-α ratio were significantly higher ( Plt;0.05) . There was no significant difference in levels of miR-150, IL-10, IL-10/TNF-α ratio, and WBC between the sepsis group and the SIRS group (Pgt;0.05) . There was no significant difference in serum concentrations of TNF-α among three groups ( Pgt;0.05) . MiR-150 expression in non-survivor sepsis patients was significantly lower than that in survivor sepsis patients (Plt;0.05) , while serum IL-10 and IL-10/TNF-αratio were significantly higher (Plt;0.01) , but there was no significant difference in serum TNF-α between the non-survivor group and the survivor group ( Pgt;0.05) . There was significantly negative correlation between miR-150 and SOFA score, TNF-α and IL-10( r=-0. 619, - 0.457, -0. 431, Plt;0.05, respectively) , but no correlation between miR-150 and WBC ( r =-0. 184, Pgt;0.05) . There was no relationship between serum TNF-α, IL-10, IL-10 /TNF-α ratio or SOFA score ( Pgt;0.05) . Conclusions MiR-150 expression in the peripheral blood specimens is significantly decreased in sepsis patients. The expression level of miR-150 not only reflect the situation of inflammatory response, but also may be used as a prognostic marker in sepsis, as it can reflect the severity of sepsis in certain degree.
Objective To study the change in serum levels of soluble CD14, tumor necrosis factor-α, E-selectin, interleukin-10 and mean arterial pressure, as well as their relationship to infection during the pathophysiologic process in endotoxemia of rabbits. Methods Sixteen rabbits were randomly divided into two groups: group A, as a control group; group B, endotoxemia group. The model of rabbit with endotoxemia were used. Endotoxin at a dose of 1.5 mg/(kg·h) or 3 mg/(kg·h) was continuously infused through external jugular vein within 2 hours, 1 hour respectively. The change of levels of serum soluble CD14, tumor necrosis factor-α, interleukin-10 and E-selectin were observed at 0 (time before infusion of endotoxin), 30, 60, 120, 180, 240, 300 minutes, while mean arterial pressure was measured by polygraphy system. Results In the group B,there was an increase of content of soluble CD14,tumor necrosis factor-α,interleukin-10 and E-selectin following 30, 120 minutes respectively,and mean arterial pressure was lower than that of group A at same time points. Conclusion The results suggest that soluble CD14,tumor necrosis factor-α,interleukin-10 and E-selectin may play an important role during the change of infection and that these changes may be closely related with severe infection.
ObjectiveTo investigate the clinical value of soluble triggering receptor expressed on myeloid cell-1 (sTREM-1) for diagnosis and prognosis of sepsis. MethodsPatients with SIRS (n=58) were divided into a sepsis group (n=40) and a non-sepsis group (n=18),and 12 healthy adults were admitted as control. Serum concentrations of sTREM-1,interleukin-6 (IL-6) and IL-10 were measured on days 1,3,7 and 14 by ELISA. According to the survival on 28th day after admission,the sepsis group was divided into survivors (n=27) and non-survivors (n=13). APACHEⅡ score and SOFA score were used to evaluate the severity of sepsis. The correlations between sTREM-1 and IL-6,IL-10,disease progression or prognosis were analyzed respectively. ResultsOn the first day of enrollment,sTREM-1,IL-6 and IL-10 [217.28(136.02-377.01) pg/mL,218.76(123.32-548.58) pg/mL and 93.86(54.23-143.1) pg/mL,respectively] in the sepsis group were significantly higher than those in the non-sepsis group [55.51(39.50-77.33) pg/mL,75.98(34.89-141.03) pg/mL and 52.49(45.66-56.72) pg/mL,respectively] and the control group [43.99(36.28-53.81) pg/mL,46.07(40.23-53.72) pg/mL and 49.79(43.31-53.14) pg/mL, respectively] (All P<0.01). For diagnosis of sepsis,the area under the curve (AUC) for sTREM-1 was 0.82 (95%CI 0.70-0.94). Levels of sTREM-1 and IL-10 in survivors of sepsis were gradually increased on 1st,3rd,7th day of enrollment,while level of sTREM-1 in non-survivors showed an obvious decrease during the observation. On the 14th of admission,sTREM-1,IL-6,IL-10 and IL-6/IL-10 ratio of non-survivors were significantly higher than those of survivors (P<0.05). There were significantly positive correlations between sTREM-1 and APACHEⅡ score,SOFA score,IL-6,IL-10 or IL-6/IL-10 ratio (r=0.624,0.454,0.407 and 0.324,respectively,all P<0.05). Logistic regression analysis indicated that serum level of sTREM-1 may be used as a prognostic factor of sepsis,but not an independent risk factor. ConclusionSerum sTREM-1 could be used as a marker to detect sepsis early,and sTREM-1 is also involved in systemic inflammatory reaction of sepsis patient and appears to be a prognostic value of sepsis.
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 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.
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 evaluate the role of interleukin-10 (IL-10) in acute pancreatitis. Methods Thirty mongrel dogs were divided into three groups based on the severity: acute edematous pancreatitis (AEP) group (n＝11), acute hemorrhagic necrotizing pancreatitis (AHNP) group (n＝12), and control group (n＝7). Serum level of IL-10 was determined with enzyme-linked immuno-sorbent assay (ELISA). Results Within 24 hours, AEP group had serum level of IL-10 significantly higher than that of AHNP group. Control group had no detectable serum IL-10. No significant difference was observed between AEP group and AHNP group at 48 hours. Conclusion The finding of low values of serum IL-10 suggests that there may be more consumption in AHNP group than in AEP group and it may be beneficial to decrease the severity of experimental acute pancreatitis.
Objective To investigate the changes and clinical significance of cytokines and inflammatory species in exhaled breath condensate (EBC) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods Thirty AECOPD patients admitted in the Department of Respiratory Medicine from March 2015 to August 2016 (smokers and passive smokers) and 21 healthy volunteers (non-smokers) were recruited in this prospective study. General information and EBC were collected from each subject. The concentrations of interleukin-17 (IL-17), IL-10, and 8-isoprestane (8-iso-PG) in EBC were measured by enzyme-linked immunosorbent assay, meanwhile lung function test was performed in the AECOPD patients. Results Both IL-17 (ng/L) and 8-iso-PG (ng/L) levels increased significantly in the AECOPD patients before and after treatment compared with the healthy controls (10.74±1.02 and 5.65±0.88 vs. 3.36±0.61, 12.35±2.25 and 9.65±1.22 vs. 6.93±1.15, P<0.05). However, IL-10 level significantly decreased in the AECOPD patients before and after treatment compared with the healthy controls (1.68±0.17 and 2.59±0.31 vs. 2.85±0.43, P<0.05). Both IL-17 and 8-iso-PG levels in the AECOPD patients were significantly lower after treatment than those before treatment (5.65±0.88 vs. 10.74±1.02, 9.65±1.22 vs. 12.35±2.25, P<0.05), but IL-10 level were significantly higher aftertreatment than those before treatment (2.59±0.31 vs. 1.68±0.17, P<0.05). FEV1, FVC, and FEV1%pred improved significantly after treatment (P<0.01). FEV1, FEV1/FVC and FEV1%pred were not significantly correlated with IL-17, IL-10 or 8-is-PG levels. Conclusion IL-17, IL-10 and 8-iso-PG may be involved in the pathogenesis of COPD, and may be important biomarkers in monitoring airway inflammation and oxide stress during the treatment of AECOPD patients.
ObjectiveTo investigate the clinical effect and mechanism of Chinese heat-clearing and phlegm-resolving recipe in the treatment of acute exacerbation of chronic obstructive pulmonary disease (AECOPD). MethodsA randomized controlled trial (RCT) was conducted. A total of sixty in-patients with AECOPD of phlegm heat and phlegm turbidity in lungs were randomly allocated to treatment group and control group with an equal number of patients between October 2009 and March 2010. The treatment duration was 10 days. Symptom scores of traditional Chinese medicine (TCM), pulmonary function, white blood cell count, polymorphonuclear neutrophils, C-reactive protein and the plasma concentration of bactericidal/permeability increasing protein (BPI) and interleukin (IL)-8 and IL-10 were detected. ResultsFor TCM syndrome of phlegm heat and phlegm turbidity, there was a statistical difference between the two groups after treatment (P<0.05). Cough, sputum amount, expectoration and coated tongue were improved obviously in the treatment group (P<0.05). The total explicit efficiency rate and effective rate were 56.0% and 84.0% in the control group and 82.1% and 92.9% in the treatment group respectively. The total explicit efficiency rate was significantly different between the two groups (P<0.05), while the total effective rate was not significantly different (P>0.05). Plasma concentration of IL-8 decreased markedly in the treatment group and IL-10 and BPI increased obviously. There was no significant difference in the change of BPI, IL-8 between the two groups (P>0.05), except for IL-10 (P<0.05). ConclusionChinese heat-clearing and phlegm-resolving recipe can improve signs and symptoms of TCM in the treatment of AECOPD (phlegm heat and phlegm turbidity in lungs), by the potential mechanism of increasing the level of IL-10.