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 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.
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.
目的 检测类风湿关节炎（RA）患者血清和关节液白细胞介素17A（IL-17A）的变化，探讨其与临床炎症指标、疾病活动性的关系。 方法 2011年6月－2012年6月采用酶联免疫吸附试验检测30例活动性RA患者和20例健康对照血清IL-17A水平，其中18例有膝关节积液RA患者同时检测配对血清和关节液IL-17A水平。 结果 RA组患者血清IL-17A水平显著高于健康对照组[（40.651 ± 16.402）、（23.799 ± 10.693） pg/mL，P＜0.05]。RA患者关节液IL-17A水平明显高于其血清中水平[（63.555 ± 23.405）、（43.727 ± 17.212） pg/mL，P＜0.05]。RA患者血清IL-17A水平只与疾病活动性评分（DAS28）呈正相关（r=0.498，P=0.020），而RA患者关节液IL-17A水平与DAS28和血清C反应蛋白有相关性（r=0.515，P=0.029；r=0.498，P=0.035）。 结论 RA患者血清和关节液IL-17A水平与疾病活动性显著相关，提示IL-17A可作为衡量疾病活动和关节损伤的标志之一。
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.
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 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.