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目的动态观察肾小球足细胞及裂孔隔膜分子nephrin,podocin和α-actinin在嘌呤霉素(puromycinaminonucleoside,PAN)大鼠肾病模型肾组织中表达的时相变化,探讨这些分子间及这些分子与蛋白尿发生的关系。方法用间接免疫荧光染色及实时定量PCR方法,检测PAN注射后12h、1d、36h、2d、5d、10d、15d及20d大鼠肾小球中nephrin,podocin和α-actinin分子分布和表达。结果(1)PAN注射后1d、2d及5d时,尿蛋白量无明显改变;10d时尿蛋白量明显增加(P=0.02);20d时恢复至对照组水平。(2)对照组大鼠肾小球中nephrin和podocin沿肾小球毛细血管袢呈连续线状分布,α-actinin沿肾小球毛细血管袢呈点线状分布。PAN注射1d后,nephrin和podocin的分布即发生改变,表现为断续、非线性分布。nephrin和podocin的分布改变随着尿蛋白的增多而加重,尿蛋白恢复时也逐渐恢复。20d时,α-actinin沿肾小球毛细血管袢呈连续线性分布。(3)免疫荧光定量分析结果表明,在PAN注射后36h(P=0.04)、2d(P=0.03)及5d(P=0.04)时,肾小球中podocin的免疫荧光染色强度明显下降,于第10d降至最低(P=0.006);自15d时逐渐恢复(P=0.007),20d后podocin的免疫荧光强度恢复至对照组水平。nephrin的免疫荧光染色强度在PAN注射第5天后出现下降(P=0.002),持续下降至第10天(P=
OBJECTIVE: To dynamically observe the phase changes of nephrin, podocin and α-actinin expression in glomerular nephropathy and nephropathy model of puromycin aminonucleoside (PAN) in rats, and to investigate the relationship between these molecules and these molecules and proteins The relationship between urine. Methods The expression and distribution of nephrin, podocin and α-actinin in glomerulus at 12h, 1d, 36h, 2d, 5d, 10d, 15d and 20d after PAN injection were detected by indirect immunofluorescence staining and real-time quantitative PCR. Results (1) At 1, 2 and 5 days after PAN injection, there was no significant change in urinary protein content. Urine protein content increased significantly at 10 days (P = 0.02), and returned to control level at 20 days. (2) The nephrin and podocin in glomeruli of rats in the control group showed a continuous linear distribution along the glomerular capillary loops, and the distribution of α-actinin along the glomerular capillaries was point-line. One day after PAN injection, the distribution of nephrin and podocin changed, showing intermittent and non-linear distribution. Changes in the distribution of nephrin and podocin increased with the increase of urinary protein, urine protein recovery gradually recovered. At 20 days, α-actinin showed a continuous linear distribution along the glomerular capillary loop. (3) Quantitative analysis of immunofluorescence showed that the intensity of immunofluorescence staining of podocin in glomerulus decreased significantly at 36h (P = 0.04), 2d (P = 0.03) and 5d (P = 0.04) The immunofluorescence intensity of podocin returned to the level of control after 20 days (P = 0.006). After 15 days, it recovered gradually (P = 0.007). The intensity of immunofluorescent staining of nephrin decreased (P = 0.002) after 5 days of PAN injection and continued to decrease until the 10th day (P =