雷公藤红素对肥胖型哮喘小鼠Th17细胞和气道炎症的影响

Effect of Celastrol on Airway Inflammation and Th17 Cells in Obese Asthmatic Mice

目的探讨雷公藤红素对肥胖型哮喘小鼠辅助性T细胞17(Th17)细胞和气道炎症的影响。方法随机将30只雄性C57BL/6小鼠分成5组,包括正常对照组、卵白蛋白(ovalbumin,OVA)致敏哮喘组、肥胖组、肥胖型哮喘组、雷公藤红素干预组。通过HE染色评估各组小鼠肺组织气道炎性细胞浸润及气道形态变化情况;免疫组织化学染色半定量分析白细胞介素(IL)-17A蛋白的表达情况;应用流式细胞术(FCM)检测小鼠脾脏Th17细胞与CD4+T淋巴细胞的比值;运用实时荧光定量聚合酶链反应法(QRT-PCR)检测各组小鼠肺组织中IL-17A mRNA的表达水平;应用酶联免疫吸附试验(ELISA)检测各组小鼠血清IL-17A水平。结果 HE染色结果显示,哮喘组、肥胖型哮喘组小鼠可见大量的炎症细胞浸润于小气道、小血管周围,肥胖组小鼠可见少量炎症性细胞浸润,雷公藤红素干预组小鼠炎症性细胞浸润明显减少,正常对照组未见炎性细胞浸润。哮喘组、肥胖组、肥胖型哮喘组小鼠肺组织中IL-17A光密度均值、IL-17A mRNA表达水平及血清中IL-17A含量显著高于正常对照组(P均<0. 01),而雷公藤红素干预组小鼠肺组织中IL-17A光密度均值、IL-17A mRNA表达水平及血清中IL-17A含量较肥胖型哮喘组显著降低,差异有统计学意义(P <0. 01)。哮喘组、肥胖组、肥胖型哮喘组小鼠脾脏分离淋巴细胞中Th17细胞/CD4+T细胞比值均较对照组明显增高,差异有统计学意义(P均<0. 01),雷公藤红素干预组小鼠Th17细胞/CD4+T细胞的比值显著低于肥胖型哮喘组,差异具有显著性(P <0. 01)。结论雷公藤红素对肥胖型哮喘小鼠体内Th17细胞及Th17细胞因子的表达有抑制作用,改善肥胖哮喘气道炎症。

OBJECTIVE To study the effect of celastrol on the airway inflammation and Th17 cells in obese asthmatic mice.METHODS Thirty male C57 BL/6 mices were randomly divided into five groups:sham group,ovalbumin(OVA)+DMSO group,diet-induced obesity(DIO)+DMSO group,DIO+OVA+DMSO group,DIO+OVA+celastrol group.H&E staining was used to examine histological changes in the lungs.Immunohistochemistry was used to observe IL-17 A protein in lung tissues;flow cytometry was used to detect the proportion of Th17 cells in CD4+T cells.The expression of IL-17 A mRNA in lung was examined by quantitative realtime RT-PCR,while concentration of cytokines IL-17 A in serum was assessed by standardized sandwich ELISA.RESULTS OVA+DMSO group and DIO+OVA+DMSO group showed significant higher inflammatory cells infiltration in the airways and small perivascular spaces of the lungs compared with sham group.DIO+DMSO group showed less inflammatory cells infiltration than DIO+OVA+DMSO group.However,DIO+OVA+celastrol group showed significantly reduced inflammatory cells infiltration.The expression of IL-17 A in lung tissues,IL-17 A mRNA in mice lung and serum IL-17 A level from DIO+OVA+DMSO group,OVA+DMSO group,and DIO+DMSO group significantly increased compared with sham group(P<0.01).However,the expression of IL-17 A in lung tissues,IL-17 A mRNA in mice lung and serum IL-17 A level significantly decreased in DIO+OVA+celastrol group compared with DIO+OVA+DMSO group(P<0.01).A significant increase of Th17 cells in CD4+T cell of mice spleen was found in OVA+DMSO group,DIO+DMSO group and DIO+OVA+DMSO group compared with sham group(P<0.01).Though,DIO+OVA+celastrol group revealed a significant reduction of Th17 cells compared with DIO+OVA+DMSO group(P<0.01).CONCLUTION Celastrol administration downregulates Th17 cells,decreases IL-17 A production in obese asthmatic mice.Celastrol effectively alleviates airway inflammation in obese asthatic mice.