泼尼松对阿霉素肾病小鼠肾脏Th17/Treg免疫平衡的影响
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摘要
目的:探讨阿霉素肾病小鼠肾脏局部Th17/Treg免疫平衡的变化及泼尼松对该模型肾脏局部Th17/Treg免疫平衡的影响。方法:6～8周龄健康雄性BALb/c小鼠,随机分为正常对照(Control)组、阿霉素肾病模型(adriamycin,ADR)组、低剂量泼尼松(prednisone,Pre)治疗[ADR+Pre(low)]组和高剂量泼尼松治疗[ADR+Pre(high)]组,两治疗组于建模第5周行泼尼松悬液灌胃(13.5 mg/kg和18 mg/kg,1次/d,共8周)。考马斯亮蓝法检测随机尿蛋白浓度;肌酐试剂盒检测随机尿肌酐浓度;肾脏组织病理HE染色及透射电镜观察肾脏病变情况;流式细胞术检测肾脏局部Th17和Treg细胞比例。结果:随建模时间进展,与Control组相比,ADR组逐渐由微小病变型肾病综合征(minimal change nephrotic syndrome,MCNS)进展为局灶节段肾小球硬化(focal segmental glomerulosclerosis,FSGS),随机尿蛋白/肌酐比值(random urinary protein-creatinine ratio,RUPCR)较对照组明显升高(均P=0.000),肾脏局部Th17明显升高(均P=0.000),Treg无明显差异,Th17/Treg逐渐失衡(均P=0.000);与ADR组相比,两泼尼松治疗组肾脏损伤均有不同程度的缓解,RUPCR均明显下降(均P<0.01),且ADR+Pre(high)组较ADR+Pre(low)组下降更为明显(P<0.05)。ADR+Pre(high)组较ADR组Th17明显下降(P<0.05),Treg无明显差异,Th17/Treg比值明显下降(均P<0.01)。结论:随阿霉素肾病小鼠肾脏Th17/Treg失衡加重,肾脏损害加重;而早期应用泼尼松治疗可通过下调肾脏局部Th17水平,延缓Th17/Treg失衡,进而发挥肾脏保护作用。
Objective:To investigate the change in Th17/Treg immunologic balance in the kidney in mice with adriamycin(ADR nephropathy and the effect of prednisone on Th17/Treg immunologic balance in the kidney in this model. Methods:Healthy male BALB/c mice aged 6-8 weeks were randomly divided into control group,ADR nephropathy group(ADR group),ADR+low-dose prednisone [ADR +Pre(low)] group,and ADR +high-dose prednisone [ADR +Pre(high)] group. Since week 5 after the model was established,the two treatment groups were given prednisone suspension by gavage at a dose of 13.5 and 18 mg/kg,respectively,once a day for 8 weeks. The Coomassie brilliant blue method was used to measure the concentration of random urinary protein;Serum Creatinine Kit was used to measure random urine creatinine;HE staining and a transmission electron microscope were used to observe renal histopathological changes;flow cytometry was used to measure the percentages of Th17 and Treg cells in the kidney. Results:Compared with the control group,the ADR group showed progression from minimal change nephrotic syndrome to focal segmental glomerulosclerosis and significant increases in random urinary protein-creatinine ratio(RUPCR)(P=0.000) and percentage of Th17 cells in the kidney(P=0.000),and there was no significant difference in the percentage of Treg between the two groups;the ADR group showed Th17/Treg imbalance(P=0.000). Compared with the ADR group,the two prednisone treatment groups had varying degrees of relief of kidney injury and a significant reduction in RUPCR(P<0.01),and the ADR+Pre(high) group had a significantly greater reduction than the ADR+Pre(low) group(P<0.05).Compared with the ADR group,the ADR+Pre(high) group had significant reductions in Th17 cells(P<0.05) and Th17/Treg ratio(P<0.01),while there was no significant difference in Treg cells between the two groups. Conclusion:Kidney injury becomes worse with the aggravation of Th17/Treg imbalance in the kidney in mice with ADR nephropathy. Early prednisone treatment can downregulate the percentage of Th17 cells in the kidney,delay Th17/Treg imbalance,and thus protect the kidney.
Objective:To investigate the change in Th17/Treg immunologic balance in the kidney in mice with adriamycin(ADR nephropathy and the effect of prednisone on Th17/Treg immunologic balance in the kidney in this model. Methods:Healthy male BALB/c mice aged 6-8 weeks were randomly divided into control group,ADR nephropathy group(ADR group),ADR+low-dose prednisone [ADR +Pre(low)] group,and ADR +high-dose prednisone [ADR +Pre(high)] group. Since week 5 after the model was established,the two treatment groups were given prednisone suspension by gavage at a dose of 13.5 and 18 mg/kg,respectively,once a day for 8 weeks. The Coomassie brilliant blue method was used to measure the concentration of random urinary protein;Serum Creatinine Kit was used to measure random urine creatinine;HE staining and a transmission electron microscope were used to observe renal histopathological changes;flow cytometry was used to measure the percentages of Th17 and Treg cells in the kidney. Results:Compared with the control group,the ADR group showed progression from minimal change nephrotic syndrome to focal segmental glomerulosclerosis and significant increases in random urinary protein-creatinine ratio(RUPCR)(P=0.000) and percentage of Th17 cells in the kidney(P=0.000),and there was no significant difference in the percentage of Treg between the two groups;the ADR group showed Th17/Treg imbalance(P=0.000). Compared with the ADR group,the two prednisone treatment groups had varying degrees of relief of kidney injury and a significant reduction in RUPCR(P<0.01),and the ADR+Pre(high) group had a significantly greater reduction than the ADR+Pre(low) group(P<0.05).Compared with the ADR group,the ADR+Pre(high) group had significant reductions in Th17 cells(P<0.05) and Th17/Treg ratio(P<0.01),while there was no significant difference in Treg cells between the two groups. Conclusion:Kidney injury becomes worse with the aggravation of Th17/Treg imbalance in the kidney in mice with ADR nephropathy. Early prednisone treatment can downregulate the percentage of Th17 cells in the kidney,delay Th17/Treg imbalance,and thus protect the kidney.
引文
[1]中华医学会儿科学分会肾脏病学组. 91所医院1990~2002年小儿慢性肾衰竭1268例调查报告[J].中华儿科杂志,2004,42(10):724-730.
[2] Kurts C,Panzer U,Anders HJ,et al. The immune system and kidney disease:basic concepts and clinical implications[J]. Nat Rev Immunol,2013,13(10):738-753.
[3] Couser WG. Basic and translational concepts of immune-mediated glomerμLar diseases[J]. J Am Soc Nephrol,2012,23(3):381-399.
[4]蓝程,杨锡强,李成荣,等. CD4+T淋巴细胞在激素敏感型肾病综合征发病中的作用研究[J].中华儿科杂志,2001,39(4):240-243.
[5] Shao XS,Yang XQ,Zhao XD,et al. The prevalence of Th17 cells and FOXP3 regμLate T cells(Treg)in children with primary nephrotic syndrome[J]. Pediatr Nephrol,2009,24(9):1683-1690.
[6] Wang L,Li Q,Wang L,et al. The role of Th17/IL-17 in the pathogenesis of primary nephrotic syndrome in children[J]. Kidney Blood Press Res,2013,37(4-5):332-345.
[7]成学琴,鲍华英,张爱华,等.儿童原发性肾病综合征不同种类糖皮质激素治疗的临床研究[J].临床儿科杂志,2013,31(2):159-161.
[8]章元沛.药理学实验[M]. 2版.北京:人民卫生出版社,1985:285.
[9] Lee VW,Harris DC. Adriamycin nephropathy:a model of focal segmental glomerμLosclerosis[J]. Nephrology(Carlton),2011,16(1):30-38.
[10]刘练,张高福,李秋,等.阿霉素肾病小鼠的肾脏病理转变过程[J].中国实验动物学报,2014,22(2):13-16.
[11] Xu Y,Lin H,Zheng W,et al. Matrine ameliorates adriamycininduced nephropathy in rats by enhancing renal function and modμ-Lating Th17/Treg balance[J]. Eur J Pharmacol,2016,791:491-501.
[12] Wang YM,Wang Y,Harris DC,et al. Adriamycin nephropathy in BALB/c mice[J]. Curr Protoc Immunol,2015,108:15.28.1-6.
[13] Maneechotesuwan K,Kasetsinsombat K,Wamanuttajinda V,et al.Statins enhance the effects of corticosteroids on the balance between regμLatory T cells and Th17 cells[J]. Clin Exp Allergy,2013,43(2):212-222.
[14]谭雄俊,杨宝辉,李秋,等. IL-10~+B细胞对儿童过敏性紫癜Th17/Treg免疫平衡紊乱的影响[J].免疫学杂志,2017,33(2):133-140.
[15] Talaat RM,Mohamed SF,Bassyouni IH,et al. Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus(SLE)patients:correlation with disease activity[J]. Cytokine,2015,72(2):146-153.
[16] Guess A,Agrawal S,Wei CC,et al. Dose-and time-dependent glucocorticoid receptor signaling in podocytes[J]. Am J Physiol Renal Physiol,2010,299(4):F845-853.
[17] Vazquez-Tello A,Semlali A,Chakir J,et al. Induction of glucocorticoid receptor-beta expression in epithelial cells of asthmatic airways by T-helper type 17 cytokines[J]. Clin Exp Allergy,2010,40(9):1312-1322.
[18] Zhou H,Tian X,Tufro A,et al. Loss of the podocyte glucocorticoid receptor exacerbates proteinuria after injury[J]. Sci Rep,2017,7(1):9833.
[19] Saksida T,Vujicic M,Nikolic I,et al. Compound A,a selective glucocorticoid receptor agonist,inhibits immunoinflammatory diabetes,induced by multiple low doses of streptozotocin in mice[J]. Br J Pharmacol,2014,171(24):5898-5909.
[2] Kurts C,Panzer U,Anders HJ,et al. The immune system and kidney disease:basic concepts and clinical implications[J]. Nat Rev Immunol,2013,13(10):738-753.
[3] Couser WG. Basic and translational concepts of immune-mediated glomerμLar diseases[J]. J Am Soc Nephrol,2012,23(3):381-399.
[4]蓝程,杨锡强,李成荣,等. CD4+T淋巴细胞在激素敏感型肾病综合征发病中的作用研究[J].中华儿科杂志,2001,39(4):240-243.
[5] Shao XS,Yang XQ,Zhao XD,et al. The prevalence of Th17 cells and FOXP3 regμLate T cells(Treg)in children with primary nephrotic syndrome[J]. Pediatr Nephrol,2009,24(9):1683-1690.
[6] Wang L,Li Q,Wang L,et al. The role of Th17/IL-17 in the pathogenesis of primary nephrotic syndrome in children[J]. Kidney Blood Press Res,2013,37(4-5):332-345.
[7]成学琴,鲍华英,张爱华,等.儿童原发性肾病综合征不同种类糖皮质激素治疗的临床研究[J].临床儿科杂志,2013,31(2):159-161.
[8]章元沛.药理学实验[M]. 2版.北京:人民卫生出版社,1985:285.
[9] Lee VW,Harris DC. Adriamycin nephropathy:a model of focal segmental glomerμLosclerosis[J]. Nephrology(Carlton),2011,16(1):30-38.
[10]刘练,张高福,李秋,等.阿霉素肾病小鼠的肾脏病理转变过程[J].中国实验动物学报,2014,22(2):13-16.
[11] Xu Y,Lin H,Zheng W,et al. Matrine ameliorates adriamycininduced nephropathy in rats by enhancing renal function and modμ-Lating Th17/Treg balance[J]. Eur J Pharmacol,2016,791:491-501.
[12] Wang YM,Wang Y,Harris DC,et al. Adriamycin nephropathy in BALB/c mice[J]. Curr Protoc Immunol,2015,108:15.28.1-6.
[13] Maneechotesuwan K,Kasetsinsombat K,Wamanuttajinda V,et al.Statins enhance the effects of corticosteroids on the balance between regμLatory T cells and Th17 cells[J]. Clin Exp Allergy,2013,43(2):212-222.
[14]谭雄俊,杨宝辉,李秋,等. IL-10~+B细胞对儿童过敏性紫癜Th17/Treg免疫平衡紊乱的影响[J].免疫学杂志,2017,33(2):133-140.
[15] Talaat RM,Mohamed SF,Bassyouni IH,et al. Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus(SLE)patients:correlation with disease activity[J]. Cytokine,2015,72(2):146-153.
[16] Guess A,Agrawal S,Wei CC,et al. Dose-and time-dependent glucocorticoid receptor signaling in podocytes[J]. Am J Physiol Renal Physiol,2010,299(4):F845-853.
[17] Vazquez-Tello A,Semlali A,Chakir J,et al. Induction of glucocorticoid receptor-beta expression in epithelial cells of asthmatic airways by T-helper type 17 cytokines[J]. Clin Exp Allergy,2010,40(9):1312-1322.
[18] Zhou H,Tian X,Tufro A,et al. Loss of the podocyte glucocorticoid receptor exacerbates proteinuria after injury[J]. Sci Rep,2017,7(1):9833.
[19] Saksida T,Vujicic M,Nikolic I,et al. Compound A,a selective glucocorticoid receptor agonist,inhibits immunoinflammatory diabetes,induced by multiple low doses of streptozotocin in mice[J]. Br J Pharmacol,2014,171(24):5898-5909.