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目的通过检测PAX2和α-平滑肌肌动蛋白(α-SMA)在单侧输尿管梗阻(UUO)大鼠肾脏组织中的表达,探讨PAX2在肾小管间质纤维化中的作用机制。方法雄性6周龄Wistar大鼠48只随机分为模型组和对照组,每组各24只。模型组在无菌条件下于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。于造模第7、14、21、28天各处死大鼠6只,取其左侧肾脏组织行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检测其肾组织PAX2mRNA表达,免疫组织化学方法检测其肾脏组织PAX2及α-SMA蛋白表达。结果1.病理结果显示:对照组各时间点肾小管及间质均无明显变化。随着梗阻时间的延长,模型组大鼠肾小管间质纤维化指数逐渐增高(Pa<0.01)。2.对照组大鼠肾组织仅有少量PAX2mRNA表达,而模型组第7天表达增高,随着梗阻时间的延长,PAX2mRNA表达逐渐增高,于第28天达最高,与对照组各时间点比较,PAX2mRNA表达有显著性差异(Pa<0.01)。3.模型组大鼠肾小管上皮细胞PAX2的蛋白表达及肾间质α-SMA的蛋白表达于第7天均增高,随梗阻时间的延长,PAX2蛋白表达及α-SMA表达逐渐增高,于第28天最高,与对照组各个时间点比较有显著性差异(Pa<0.01)。4.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=0.880P<0.05),PAX2蛋白及α-SMA的表达均与肾小管间质纤维化指数均呈显著正相关(r=0.886,0.918Pa<0.05)。结论PAX2可能参与UUO大鼠模型肾小管上皮细胞-肌成纤维细胞转分化,使间质中肌成纤维细胞的数量增多,引起细胞外基质积聚增加,导致了肾小管间质纤维化的发生发展。
Objective To investigate the mechanism of PAX2 in tubulointerstitial fibrosis by detecting the expression of PAX2 and α-smooth muscle actin (α-SMA) in the kidney of rats with unilateral ureteral obstruction (UUO). Methods 48 male Wistar rats of 6 weeks old were randomly divided into model group and control group, 24 in each group. The model group underwent a ureter ligation on the left side of the left dorsal rib under aseptic conditions while the control group only examined the renal capsule and did not ligate the ureter. At the 7th, 14th, 21st and 28th day after modeling, 6 rats were sacrificed, and the left renal tissues were taken out to detect the pathological changes of the kidney by HE and Masson staining to 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. Pathological results showed that: the control group at each time point tubules and interstitial no significant change. With the prolongation of obstruction, the index of tubulointerstitial fibrosis gradually increased in model group (Pa <0.01). The expression of PAX2mRNA in the control group rats was only a small amount, but the expression of PAX2mRNA in the model group increased on the7th day. With the extension of the obstruction time, the expression of PAX2mRNA increased gradually and reached the highest on the28th day. Compared with the control group, PAX2 mRNA expression was significantly different (Pa <0.01). The protein expression of PAX2 and the expression of α-SMA in renal tubular epithelial cells in model group increased on the 7th day, and the expression of PAX2 and α-SMA gradually increased with the prolongation of obstruction. 28 days the highest, with the control group at all time points were significantly different (Pa <0.01). The expression of PAX2 protein in renal tubular epithelial cells of model group was positively correlated with the expression of α-SMA in renal interstitium (r = 0.880P <0.05), the expressions of PAX2 protein and α-SMA were correlated with tubulointerstitial fibrosis Index were significantly correlated (r = 0.886,0.918Pa <0.05). Conclusions PAX2 may be involved in the transdifferentiation of renal tubular epithelial cells - myofibroblasts in UUO rat model, resulting in an increase in the number of interstitial myofibroblasts, resulting in an increase of extracellular matrix accumulation, leading to the development of tubulointerstitial fibrosis .