摘要
目的观察肾移植术后急性排斥大鼠外周血中叉头状转录因子3(Foxp3)阳性的CD4^+CD25^+调节性T细胞(CD4^+CD25^+ Treg细胞)水平及其T盒转录因子TBX1(TBX1)和肌细胞特异性增强因子2D(MEF2D)表达变化,结合肾功能、血浆白细胞介素10(IL-10)、干扰素γ(IFN-γ)及肾组织病理变化,探讨其中的调控机制。方法建立大鼠肾移植模型,分为急性排斥组(急排组)和非急性排斥(非急排)组。测定两组大鼠肾功能(尿素氮和血肌酐);ELISA法测定血浆中IL-10和IFN-γ;HE染色法观察肾脏组织病理。应用流式细胞术分选测定CD4^+CD25^+ Treg细胞;实时定量PCR测定CD4^+CD25^+ Treg细胞的Foxp3、TBX1和MEF2D mRNA表达;Western印迹测定CD4^+CD25^+ Treg细胞Foxp3、TBX1和MEF2D蛋白表达。结果与非急排组相比,急排组尿素氮和血肌酐均升高,IL-10下降,IFN-γ升高(均P<0.05)。急排组肾组织病理出现肾小球肥大和系膜细胞显著增生,毛细血管增生和中性粒细胞浸润;肾间质明显水肿和肾小管坏死,伴有淋巴细胞、浆细胞及中性粒细胞浸润。与非急排组相比,急排组外周血CD4^+CD25^+ Treg细胞比例降低(4.50%±0.50%比5.74%±1.96%,P<0.05),CD4^+CD25^+ Treg细胞Foxp3、MEF2D的mRNA和蛋白表达均降低(均P<0.01),TBX1的mRNA和蛋白表达均升高(均P<0.01)。结论肾移植急性排斥大鼠免疫耐受细胞CD4^+CD25^+ Treg细胞减少,Foxp3、MEF2D、IL-10表达降低,TBX1、IFN-γ表达增强,共同促进肾移植急性排斥反应的发生发展,加重肾损伤。
Objective To observe the level of CD4^+CD25^+ regulatory T cells (CD4^+CD25^+ Treg cells) with positive fork head transcription factor 3 (Foxp3) and changes of T-box transcription factor TBX1 (TBX1) and myocyte specific enhancer 2D (MEF2D) expression in peripheral blood of rats with acute rejection after renal transplantation, and to investigate its regulatory mechanisms by combined with renal function, plasma interleukin-10 (IL-10), interferon-γ(IFN-γ) and renal histopathological changes. Methods Rat renal transplantation model was established and divided into two groups: acute rejection group (AR group) and non-acute rejection group (non-AR group). Their renal function including serum creatinine (Scr) and blood urea nitrogen (BUN) in plasma was measured. The renal histopathology was observed by HE staining. Levels of IL-10 and IFN-γ in plasma were detected by ELISA. The proportion of CD4^+CD25^+ Treg cells was measured by flow cytometry. The mRNA expressions of Foxp3, TBX1 and MEF2D in CD4^+CD25^+ Treg cells were detected by real-time PCR, and their protein expressions were tested by Western blotting. Results Compared with those in the non-AR group, the levels of BUN, Scr and IFN-γ significantly increased in AR group (all P<0.05), while IL-10 decreased (P<0.05). Renal histopathology in the acute rejection group showed glomerular hypertrophy and mesangial cell proliferation, capillary proliferation and neutrophil infiltration;renal interstitial edema and tubular necrosis, accompanied by lymphocytes, plasma cells and neutrophils infiltration. Compared with that in the non-AR group, the percentage of CD4^+CD25^+ Treg cells in peripheral blood was notably lowered in AR group (4.50%±0.50% vs 5.74%±1.96%, P<0.05). The mRNA and protein expressions of Foxp3 and MEF2D were lower in AR group than those in non-AR group, while the expressions of TBX1 was elevated (all P<0.05). Conclusions In rats with acute renal allograft rejection, the percentage of CD4^+CD25^+ Treg cells and expressions of Foxp3, MEF2D and IL-10 decrease, while the expressions of TBX1 and IFN-γ enhance. These participate in the development of acute rejection after renal transplantation, and aggravate the renal damage.
作者
袁树珍
许云鹏
隋晓露
顾凤娟
张艾莎
张燕子
谢婷妃
陈继红
Yuan Shuzhen;Xu Yunpeng;Sui Xiaolu;Gu Fengjuan;Zhang Aisha;Zhang Yanzi;Xie Tingfei;Chen Jihong(Department of Nephrology, Bao an People's Hospital, Shenzhen 518100, China)
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2019年第4期295-301,共7页
Chinese Journal of Nephrology
