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[目的]通过检测全长配对盒基因2(airedbox2,PAX2)和平滑肌肌动蛋白(α-smooth muscle actin,SMA)在单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾脏组织中的表达,初步探讨PAX2在肾小管间质纤维化中的作用机制。[方法]24只雄性6周龄Wistar大鼠随机分为模型组和对照组,每组12只。模型组在无菌条件下左侧背部肋下切口行左侧输尿管结扎术,对照组只探及肾包膜,不结扎输尿管。于造模后d21处死大鼠,检测血清肌酐(serum creatinine,Scr)和尿素氮(blood urea nitrogen,BUN)水平。取左侧肾脏组织做HE和Masson染色检测肾脏病理改变,计算肾小管间质纤维化指数。采用实时荧光定量PCR方法检测肾组织中PAX2 mRNA表达,免疫组织化学方法检测肾脏组织PAX2及α-SMA蛋白的表达。[结果]与假手术组比较,模型组大鼠BUN、Scr水平显著增高(P﹤0.01);肾小管间质纤维化指数显著增高(P﹤0.01)。模型组大鼠肾小管上皮细胞PAX2的蛋白以及肾间质α-SMA的蛋白表达均较对照组显著增高(P﹤0.01)。对照组大鼠肾组织中仅有少量PAX2mRNA表达,模型组PAX2的mRNA表达显著增高(P﹤0.01)。直线相关分析显示:模型组大鼠肾小管上皮细胞PAX2蛋白表达与肾间质α-SMA表达呈显著正相关(r=0.882,P﹤0.05);与肾小管间质纤维化指数BUN、Scr分别均呈显著正相关(r=0.899,0.731,0.933均P﹤0.05)。[结论]在UUO大鼠模型中,肾脏组织PAX2和α-SMA表达均显著增加。PAX2可能参与肾小管上皮细胞-肌成纤维细胞转分化,使间质中肌成纤维细胞的数量增多,引起细胞外基质(Extracellular Matrix,ECM)积聚增加,导致了小管肾间质纤维化的发生发展。
[Objective] To investigate the expression of airedbox2 (PAX2) and α-smooth muscle actin (SMA) in the kidney of unilateral ureteral obstruction (UUO) rats, To investigate the mechanism of PAX2 in tubulointerstitial fibrosis. [Methods] Twenty-four male Wistar rats, 6 weeks old, were randomly divided into model group and control group, with 12 rats in each group. The model group underwent left ureteral ligation of the left dorsal rib under aseptic conditions, while the control group only probed into the renal capsule without ligation of the ureter. The rats were killed at d21 after the model was established, and serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured. Take the left kidney tissue HE and Masson staining to detect renal pathological changes, calculate the tubulointerstitial fibrosis index. The expression of PAX2 mRNA in renal tissues was detected by real-time fluorescence quantitative PCR, and the expression of PAX2 and α-SMA protein in renal tissues was detected by immunohistochemistry. [Results] The levels of BUN and Scr in model group were significantly higher than those in sham operation group (P <0.01), and the index of tubulointerstitial fibrosis was significantly increased (P <0.01). The expression of PAX2 protein in renal tubular epithelial cells and the expression of α-SMA in renal interstitium in model group were significantly higher than those in control group (P <0.01). There was only a small amount of PAX2 mRNA expression in the kidney of the control group, and the mRNA expression of PAX2 in the model group was significantly increased (P <0.01). Linear correlation analysis showed that there was a significant positive correlation between PAX2 protein expression and renal interstitial α-SMA expression in model group (r = 0.882, P <0.05), and tubulointerstitial fibrosis index BUN and Scr There was a significant positive correlation (r = 0.899,0.731, 0.933, P <0.05). [Conclusion] The expressions of PAX2 and α-SMA in kidney tissue were significantly increased in UUO rat model. PAX2 may be involved in the transdifferentiation of renal tubular epithelial cells - myofibroblasts, increasing the number of myofibroblasts in the stroma, resulting in the accumulation of extracellular matrix (ECM), resulting in the occurrence of tubulointerstitial fibrosis development of.