Objective To investigate the role of T helper 17 ( Th17) cells and CD4 + CD25 + Foxp3+regulatory T cells ( Treg) in the pathogenesis of asthma in a mouse model. Methods Twenty-four BALB/ c mice were randomly divided into an asthma group and a normal control group, with 12 mice in each group.Asthma model was established by ovalbumin sensitization and aerosol challenge in the asthma group. Airway reactivity was measured by plethysmography. The total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were measured. The ratio of Th17 and Treg cells to mononuclear cells in the spleens of mice were detected by flow cytometry. The levels of IL-17 and IL-10 in BALF and lung homogenates were measured by ELISA. Results The bronchial provocation test showed that the average lung resistance increased remarkably in the asthma group. In spleens of the asthmatic mice, the percentage of Th17 cells was significantly higher [ ( 5.68 ±1. 99)% vs ( 2.80 ±0. 82) %, P lt; 0. 01] , and the percentage of Treg cells was significantly lower [ (2.88 ±0. 46) % vs ( 6.10 ±2.44) % , Plt; 0. 01] , with the ratio of Th17 to Treg significantly increased( 1. 93 ±0. 41 vs 0. 50 ±0. 15,P lt;0. 01) . In BALF and lung homogenates of the asthma group, the level of IL-17 was significantly higher[ ( 22. 37 ±3. 00) pg/mL vs ( 11. 42 ±2. 15) pg/mL, ( 52. 93 ±5. 39) pg/mL vs ( 19. 38 ±2. 65) pg/mL, both Plt; 0. 01] , and the level of IL-10 was significantly lower[ ( 6. 05 ±1. 25) pg/mL vs ( 14. 23 ±2. 94) pg/mL, ( 9. 33 ±1. 79) pg/mL vs ( 21. 40 ±2. 44) pg/mL, both P lt; 0. 01] compared with the control group.Conclusion The imbalance of Th17/ Treg plays an important role in the pathogenesis of asthma.
Objective To investigate the changes of interleukin-17 ( IL-17) and the effects of propofol in rats with acute lung injury ( ALI) . Methods ALI model was established by hydrochloric acid ( HCl) inhalation in a dose of 2 mL/kg. 35 adultmale SD rats were randomly divided into seven groups, ie.a control group, a HCl group, and five propofol groups ( T24b , T12b , T0 , T1a , T3a groups, respectively) . The T0 ,T24b and T12b groups were pretreated with intraperitoneal propofol injection 0, 24 and 12 hours respectively before HCl inhalation. The T1a and T3a groups were managed by intraperitoneal propofol injection 1 and 3 hours respectively after HCl inhalation. Immunohistochemistry was used to determine the expression of IL-17 in lung tissue. ELISA was adopted to detect the levels of IL-17 and IL-8 in lung tissue homogenate as well as in bronchoalveolar lavage fluid ( BALF) , meanwhile arterial partial pressure of oxygen ( PaO2 ) and myeloperoxidase ( MPO) were measured. Results Those rats in the HCl group appeared respiratory distress, cyanosis, pulmonary edema, and inflammatory cells infiltration in lung tissues after HCl inhalation.The IL-17 levels in lung tissue homogenate as well as in BALF were higher in the HCl group than those in the control group( all P lt; 0. 01) . IL-17 was mainly expressed in alveolar epithelial cells and mononuclear cells in the ALI rats and its expression level was higher than that in the control group. IL-17 concentration in lung tissue homogenate was both correlated with IL-8 concentration in lung tissue homogenate ( r=0. 98, P =0.003) and with the activity of MPO in lung tissue( r=0. 981, P =0. 003) in the HCl group. Mainwhile, a same significant correlation was found between IL-8 level in lung tissue homogenate and the MPO activity in the HCl group( r =0. 961, P =0. 009) . Propofol attenuated lung injury induced by HCl inhalation, especially in T24b group. The concentrations of IL-17 in lung tissue homogenate and in BALF were lower in T24b group when compared with the HCl group( P = 0. 011, P =0. 003, respectively) . Conclusions The expression of IL-17 increases in ALI rats. Pretreatment with propofol by 24 hours has obvious inhibiting effects on inflammatory reaction. Inhibiting IL-17 expression may be one of the mechanisms through which propofol inhibits the inflammatory reaction of ALI.
【Abstract】 Objective To observe the plasma levels of adiponectin and interleukin-17 ( IL-17) in patients with chronic obstructive pulmonary disease ( COPD) at acute exacerbation or stable stage, and analyze their relationship. Methods Sixty male COPD patients with normal weight ( with BMI range of 18. 5-24. 9 kg/m2 ) were enrolled, including 30 patients with acute exacerbations of COPD ( AECOPD) and 30 patients with stable COPD. Twenty healthy nonsmoking male volunteers were included as controls. The plasma levels of adiponectin and IL-17 as well as lung function ( FEV1% pred and RV% pred) were measured in all subjects. Results The concentrations of adiponectin and IL-17 were significantly higher in the AECOPD patients than those of the patients with stable COPD and the contro1s ( P lt; 0. 001) . Theconcentrations of adiponectin and IL-17 were significantly higher in the patients with stable COPD than those of the controls ( P lt;0. 01) . Adiponectin was positively correlated with IL-17 in the AECOPD patients ( r =0. 822, P lt;0. 001) and in the patients with stable COPD ( r =0. 732, P lt;0. 001) . Adiponectin was positivelycorrelated with RV% pred in the AECOPD patients ( rs = 0. 764, P lt;0. 001) and in the patients with stable COPD ( rs =0. 967, P lt;0. 001) . There was no significant relationship between adiponectin and FEV1% pred ( P gt;0. 05) . Conclusions The plasma level of adiponectin in COPD patients is elevated which is relatedwith excessive inflation of lung. Adiponectin may be involved in the process of inflammation in COPD as a new pro-inflammatory cytokine.
Objective To investigate the expressions of ubiquitin-proteasome markers，including E2-14K，MAFbx，MuRF-1，and nuclear factor-κB（NF- κB） p50，in diaphragm of COPD rats，and their relationship with IL-17 level in diaphragm and serum in order to elucidate the potential mechanism of diaphragm atrophy. Methods Thirty healthy adult male SD rats were randomly divided into a model group （n=18） and a normal control group （n=12）. The COPD rat model was established by instillation of lipopolysaccharide （LPS） and exposure to cigarette smoke for 28 days. The protein levels of E2-14K，MAFbx，MuRF-1，and NF-κB p50 in diaphragm were measured by Western blot. The concentration of IL-17 in serum and diaphragm was measured by ELISA. Results Western blot showed that the protein expressions of E2-14K，MAFbx，MuRF-1，and NF-κB p50 in diaphragm increased significantly in the COPD model group compared with the normal control group （0.96±0.12 vs. 0.53±0.09，0.99±0.10 vs. 0.53±0.08，0.95±0.08 vs. 0.51±0.16，1.11±0.10 vs. 0.64±0.50，respectively，Plt;0.01）. The IL-17 level in serum and diaphragm was significantly higher in the COPD group than the control group. The expression of NF-κB p50 was positively correlated with E2-14K，MAFbx，and MuRF-1 expressions （r=0.82，0.92，0.86，respectively，Plt;0.01）. Both in serum and diaphragm，IL-17 level was positively correlated with the percentage of neutrophils，levels of NF-κB p50，E2-14K，MAFbx，and MuRF-1 expressions（all Plt;0.01）. The IL-17 levels in serum and diaphragm were also positively correlated each other （r=0.84，Plt;0.01）. Conclusions The results show that the ubiquitin-proteasom pathway，the NF-κB pathway and IL-17 are up-regulated in diaphragm of COPD rats .These alterations may contribute to diaphragm atrophy in COPD.
Objective To analyze the relationship of serum IL-17, IL-8 levels and BODE index in patients with stable COPD, and investigate the clinical value of serum IL-17 and IL-8 in evaluating disease severity and prognosis. Methods A comparative study was performed in40 clinically stable COPD patients and 40 matched healthy individuals. The serum levels of IL-17 and IL-8 in both groups were measured. Correlation analysis was performed between serum IL-17, IL-8 levels and BODE index in the patients with stable COPD. Results The serumlevels of IL-17 and IL-8 in the COPD group were ( 114. 02 ±34. 84) pg/mL and ( 102. 67 ±31. 55) pg/mL, increased significantly compared with those in the healthy group which were ( 73. 22 ±14. 66) pg/mL and ( 35. 36 ±5. 04) pg/mL ( P lt;0. 05) respectively. Both of serum IL-17 and IL-8 levels were positively correlated with BODE index in the patients with stable COPD ( r = 0. 782, r = 0. 924, P lt;0. 001) . Conclusions High levels of serumIL-17 and IL-8 implies active inflammation in patients with stable COPD. Detection of serumIL-17 and IL-8 can help evaluate disease severity and prognosis.
Objective To investigate the expression and significance of Fork head /winged helix protein 3 (Foxp3) , retinoic acid-related orphan receptorγt (RORγt) , and interleukin-17 (IL-17) in Guinea pigs with emphysema. Methods Smoking and active immunization with elastin were separately used in guinea pigs to establish emphysema model. Then the destruction of lung tissue was assayed by measurement of the average radius of alveolar. The expressions of Foxp3 , RORγt, and IL-17 in lung tissue of the guinea pigs were detected by immunohistochemical technique. The results were compared with the normal control group by the analysis of variance or kruskal-Wallis test. Spearman rank correlation was used to analyze the correlation between the ratio of Foxp3/RORγt and IL-17, also the correlation between Foxp3/RORγt and the average radius of alveolar. Results In the smoking group and the active immunization group, the average radius of alveolar were significantly longer than the control group (Plt;0.05) . And the expression of Foxp3/RORγt was significantly unbalanced, with the number of Foxp3-positive cells decreased and RORγt-positive cells increased (Plt;0.05) . Meanwhile the level of IL-17 was significantly increased compared with the control group ( Plt;0.05) . The difference between the smoking group and the active immunization group was not significant (Pgt;0.05) . The ratio of Foxp3/RORγt was negatively correlated with the level of IL-17 and the average radius of alveolar. Conclusions Active immunization with elastin can induce emphysema in guinea pigs. The Foxp3/RORγt expression was unbalanced in lung tissue of guinea pigs with emphysema.This imbalance may be an important mechanism attributed to the disordered expression of CD4+ Treg cells and Th17 cells, which may be involved in autoimmune regulation and development of chronic obstructive pulmonary disease.
ObjectiveTo detect expressions of interleukin-17 (IL-17) and interleukin-27 (IL-27) proteins in liver tissue of hepatic alveolar echinococcosis. MethodsThe edge liver tissues (from the lesion edge 0.5 cm) and the normal liver tissues (from the lesion edge 5 cm) of 20 patients with hepatic alveolar echinococcosis in the Affiliated Hospital of Qinghai University were collected and stored at-80℃ freezer. The immunohistochemical method was used to detect the expressions of IL-17 and IL-27 proteins in these two tissues. ResultThe positive rates of IL-17 and IL-27 protein expressions in the edge liver tissues were significantly higher than those in the normal liver tissues[IL-17:80.0% (16/20) versus 10.0% (2/20), χ2=12.36, P < 0.01; IL-27:85.0% (17/20) versus 20.0% (4/20), χ2=12.36, P < 0.01]. ConclusionHigh expressions of IL-17 and IL-27 protein in edge liver tissue might participate in progress of hepatic alveolar echinococcosis.
ObjectiveTo address the effect and mechanism of interleukin 17 (IL-17) on the proliferation, migration and apoptosis of human retinal vascular endothelial cells (HREC). MethodsIL-17 receptor (IL-17R) mRNA and protein expression in human retinal vascular endothelial cells (HREC) were quantified by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation of HREC was examined using CCK-8 assay in the presence of different concentrations of IL-17. Cell migration of HREC was detected using wound scratch assay. Flow cytometry was used to test the effect of IL-17 on the apoptosis of HREC. The effects of IL-17 on HREC expression of basic fibroblast growth factor (bFGF), Caspase-3 and thrombospondin-1 (TSP-1) were quantified by reverse transcriptase polymerase chain reaction (RT-PCR). The effect of IL-17 on HREC expression of Caspase-3 was examined using Western blot. ResultsIL-17 receptor (IL-17R) expressed in HREC as quantified by RT-PCR and Western blot. The proliferation of HREC in the presence of IL-17 was promoted in a dosage-dependent manner (t=-3.235, -6.276;P=0.032, 0.000). Wound scratch assay showed a significant increase in the migrated distance of HREC with IL-17 stimulation under the concentration of 100μg/L(t=-3.551, -2.849; P=0.006, 0.019), 200μg/L(t=-10.347, -4.519; P=0.000, 0.001) and 500μg/L (t=-3.541, -2.607; P=0.008, 0.036). The intervention of 200μg/L IL-17 can effectively inhibit the apoptosis of HREC, compared with the control group using flow cytometry (t=5.682, P=0.047). RT-PCR results showed that IL-17 can promote the expression of bFGF and inhibit the expression of Caspase-3 and TSP-1. Western blot result also showed that IL-17 can suppress the protein expression of Caspase-3. ConclusionThe mechanism of IL-17 promoting proliferation, migration but suppress apoptosis of HREC may via regulating the expression of bFGF and Caspase-3.
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.
Objective To observe the effect of celastrol on the secretion of interleukin (IL)-17 in peripheral blood mononuclear cells in patients with sympathetic ophthalmia (SO), and its possible mechanisms. Methods Venous blood samples were extracted from 10 cases of sympathetic ophthalmia patients and 10 health objectives. The peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation and then were divided into 4 groups. Group A (control group): PBMCs of health objectives; Group B: PBMCs of SO patients; Group C: PBMCs of SO patients with 0.5 μmol/L celastrol in the medium; Group D: PBMCs of SO patients with 1 μmol/L celastrol in the medium. After culturing the cells for 3 days, the supernatant of 4 groups was collected, and the levels of IL-23 and IL-17 were detected by enzyme-linked immuno sorbent assay (ELISA). Then, the 50 ng/ml rIL-23 was added into the medium of group A which was the group A1; the 50ng/ml rIL-23 and 1 μmol/L Cela were added into to the medium of group A which was the group A2. For the medium of group B, the 50 ng/ml rIL-23 was added into the medium which was the group B1; the 50 ng/ml rIL-23 and 1 μmol/L celastrol were added into to the medium of group B which was the group B2. After culturing for 3 days, the supernatant of cells of these 4 groups was collected, and the levels of IL-17 were detected by ELISA. Results In group A, the levels of IL-23 and IL-17 were (228.43±17.27) pg/ml and (220.55±31.15) pg/ml respectively. In group B, the levels of IL-23 and IL-17 were (513.85±36.46) pg/ml and (866.77±72.92) pg/ml respectively. In group C, the levels of IL-23 and IL-17 were (381.07±20.93) pg/ml and (517.43±54.87) pg/ml respectively. In group D, the levels of IL-23 and IL-17 were (237.14±17.97) pg/ml and (242.89±34.09) pg/ml respectively. Between group A and D, there was no statistically significant difference in IL-23 or IL-17 level (P＞0.05); but when comparing other groups, the differences were statistically significant (P＜0.05). The levels of IL-17 in group A1 and group A2 were (428.43±24.53) pg/ml and (229.15±23.28) pg/ml and the difference was statistically significant (P＜0.05). The levels of IL-17 in group B1 and group B2 were (1373.39±89.51) pg/ml and (571.01±94.88) pg/ml and the difference was statistically significant (P＜0.05). Conclusion Celastrol can inhibit the secretion of IL-17 by PBMCs in SO patients via inhibiting the secretion of IL-23.