泼尼松对阿霉素肾病小鼠肾脏Th17/Treg免疫平衡的影响

Effect of prednisone on Th17/Treg immunologic balance in mice with adriamycin nephropathy

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 seg－mental 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。

Abstract:

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 pred－nisone[ADR+Pre（low）] group，and ADR+high-dose prednisone[ADR+Pre（high）] group. Since week 5 after the model was estab－lished，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 glomeruloscle－rosis 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 vary－ing 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 downregu－late the percentage of Th17 cells in the kidney，delay Th17/Treg imbalance，and thus protect the kidney.