雷公藤红素对肥胖型哮喘小鼠Th17细胞和气道炎症的影响
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摘要
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.
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.
引文
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[11] ENDO Y,YOKOTE K,NAKAYAMA T,et al. The obesity-related pathology and Th17 cells[J]. Cell Mol Life Sci,2017,74(7):1231-1245.
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[16] SCHINDLER T I,WAGMER J J,GOEDICKE-FRITZ S,et al.TH17 Cell Frequency in Peripheral Blood Is Elevated in Overweight Children without Chronic Inflammatory Diseases[J].Front Immunol,2017,8:1543.
[17] BETTELLI E,KORN T,KUCHROO V K. Th17:the third member of the effector T cell trilogy[J]. Curr Opin Immunol,2007,19(6):652-657.
[18] KANELLOPOULOU C,MULJO S A. Fine-tuning Th17 cells:to be or not to be pathogenic[J]. Immunity,2016,44(6):1241-1243.
[19] LI K,WANG Z,CAO Y,et al. The study of the ratio and distribution of Th17 cells and Tc17 cells in asthmatic patients and the mouse model[J]. Asian Pac J Allergy Immunol,2013,31(2):125-131.
[20] ELAIDY S M,ESSAWY S S,HUSSAIN M A,et al. Modulation of the IL-23/IL-17 axis by fenofibrate ameliorates the ovalbumin/lipopolysaccharide-induced airway inflammation and bronchial asthma in rats[J]. Naunyn Schmiedebergs Arch Pharmacol,2018,391(3):309-321.
[21] JOEAN O,HUEBER A,FELLER F,et al. Suppression of Th17-polarized airway inflammation by rapamycin[J]. Sci Rep,2017,7(1):15336.
[22] LIU C,ZHU L,FUKUDA K,et al. The flavonoid cyanidin blocks binding of the cytokine interleukin-17A to the IL-17RA subunit to alleviate inflammation in vivo[J]. Sci Signal,2017,10(467). pii:eaaf8823.
[23] ZHANG W,ZHANG X,SHENG A,et al.γ-Secretase gammaSecretase Inhibitor Alleviates Acute Airway Inflammation of Allergic Asthma in Mice by Downregulating Th17 Cell Differentiation[J]. Mediators Inflamm,2015,2015,258168.
[24] MCKINLEY L,ALCORN J F,PETERSON A,et al. TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice[J]. J Immunol,2008,181(6):4089-4097.
[25] ZHANG J,SHAN J,CHEN X,et al. Celastrol mediates Th17and Treg cell generation via metabolic signaling[J]. Biochem Biophys Res Commun,2018,497(3):883-889.
[26] ASTRY B,VENKATESHA S H,LAURENCE A,et al. Celastrol,a Chinese herbal compound,controls autoimmune inflammation by altering the balance of pathogenic and regulatory T cells in the target organ[J]. Clin Immunol,2015,157(2):228-238.
[27] WANG Y,CAO L,XU L M,et al. Celastrol Ameliorates EAE Induction by Suppressing Pathogenic T Cell Responses in the Peripheral and Central Nervous Systems[J]. J Neuroimmune Pharmacol,2015,10(3):506-516.
[2] WENZEL S E. Asthma phenotypes:the evolution from clinical to molecular approaches[J]. Nat Med,2012,18(5):716-725.
[3] ANO S,MORISHIMA Y,ISHII Y,et al. Transcription factors GATA-3 and RORgammat are important for determining the phenotype of allergic airway inflammation in a murine model of asthma[J]. J Immunol,2013,190(3):1056-1065.
[4] MORISHIMA Y,ANO S,ISHII Y,et al. Th17-associated cytokines as a therapeutic target for steroid-insensitive asthma[J].Clin Dev Immunol,2013,2013:609395.
[5] ENDO Y,ASOU H K,MATSUGAE N,et al. Obesity Drives Th17 Cell Differentiation by Inducing the Lipid Metabolic Kinase,ACC1[J]. Cell Rep,2015,12(6):1042-1055.
[6] KIM D Y,PARK J W,JEOUNG D,et al. Celastrol suppresses allergen-induced airway inflammation in a mouse allergic asthma model[J]. Eur J Pharmacol,2009,612(1-3):98-105.
[7] LIU J,LEE J,SALAZAR HERNANDEZ M A,et al. Treatment of obesity with celastrol[J]. Cell,2015,161(5):999-1011.
[8] KIM Y,KIM K,LEE H,et al. Celastrol binds to ERK and inhibits FcepsilonRI signaling to exert an anti-allergic effect[J].Eur J Pharmacol,2009,612(1-3):131-142.
[9] WINER S,PALTSER G,CHAN Y,et al. Obesity predisposes to Th17 bias[J]. Eur J Immunol,2009,39(9):2629-2635.
[10] LIANG L,HUR J,KANG J Y,et al. Effect of the anti-IL-17antibody on allergic inflammation in an obesity-related asthma model[J]. Korean J Intern Med,2018,33(6):1220-1223.
[11] ENDO Y,YOKOTE K,NAKAYAMA T,et al. The obesity-related pathology and Th17 cells[J]. Cell Mol Life Sci,2017,74(7):1231-1245.
[12] LUCZYNSKI W,GRUBCZAK K,MONIUSZKO M,et al. Elevated levels of Th17 cells in children with central obesity[J].Scand J Clin Lab Invest,2015,75(7):595-601.
[13] SUMARAC-DUMANOVIC M,STEVANOVIC D,LIUBIC A,et al. Increased activity of interleukin-23/interleukin-17 proinflammatory axis in obese women[J]. Int J Obes(Lond),2009,33(1):151-156.
[14] MCLAUGHLIN T,LIU L F,LAMENDOLA C,et al. T-cell profile in adipose tissue is associated with insulin resistance and systemic inflammation in humans[J]. Arterioscler Thromb Vasc Biol,2014,34(12):2637-2643.
[15] YAN X,HUANG Y,WANG H,et al. Maternal obesity induces sustained inflammation in both fetal and offspring large intestine of sheep[J]. Inflamm Bowel Dis,2011,17(7):1513-1522.
[16] SCHINDLER T I,WAGMER J J,GOEDICKE-FRITZ S,et al.TH17 Cell Frequency in Peripheral Blood Is Elevated in Overweight Children without Chronic Inflammatory Diseases[J].Front Immunol,2017,8:1543.
[17] BETTELLI E,KORN T,KUCHROO V K. Th17:the third member of the effector T cell trilogy[J]. Curr Opin Immunol,2007,19(6):652-657.
[18] KANELLOPOULOU C,MULJO S A. Fine-tuning Th17 cells:to be or not to be pathogenic[J]. Immunity,2016,44(6):1241-1243.
[19] LI K,WANG Z,CAO Y,et al. The study of the ratio and distribution of Th17 cells and Tc17 cells in asthmatic patients and the mouse model[J]. Asian Pac J Allergy Immunol,2013,31(2):125-131.
[20] ELAIDY S M,ESSAWY S S,HUSSAIN M A,et al. Modulation of the IL-23/IL-17 axis by fenofibrate ameliorates the ovalbumin/lipopolysaccharide-induced airway inflammation and bronchial asthma in rats[J]. Naunyn Schmiedebergs Arch Pharmacol,2018,391(3):309-321.
[21] JOEAN O,HUEBER A,FELLER F,et al. Suppression of Th17-polarized airway inflammation by rapamycin[J]. Sci Rep,2017,7(1):15336.
[22] LIU C,ZHU L,FUKUDA K,et al. The flavonoid cyanidin blocks binding of the cytokine interleukin-17A to the IL-17RA subunit to alleviate inflammation in vivo[J]. Sci Signal,2017,10(467). pii:eaaf8823.
[23] ZHANG W,ZHANG X,SHENG A,et al.γ-Secretase gammaSecretase Inhibitor Alleviates Acute Airway Inflammation of Allergic Asthma in Mice by Downregulating Th17 Cell Differentiation[J]. Mediators Inflamm,2015,2015,258168.
[24] MCKINLEY L,ALCORN J F,PETERSON A,et al. TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice[J]. J Immunol,2008,181(6):4089-4097.
[25] ZHANG J,SHAN J,CHEN X,et al. Celastrol mediates Th17and Treg cell generation via metabolic signaling[J]. Biochem Biophys Res Commun,2018,497(3):883-889.
[26] ASTRY B,VENKATESHA S H,LAURENCE A,et al. Celastrol,a Chinese herbal compound,controls autoimmune inflammation by altering the balance of pathogenic and regulatory T cells in the target organ[J]. Clin Immunol,2015,157(2):228-238.
[27] WANG Y,CAO L,XU L M,et al. Celastrol Ameliorates EAE Induction by Suppressing Pathogenic T Cell Responses in the Peripheral and Central Nervous Systems[J]. J Neuroimmune Pharmacol,2015,10(3):506-516.
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