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目的动态观察阿霉素肾病大鼠在发生蛋白尿的过程中肾组织血管生成素样蛋白3(ANGPTL3)mRNA表达水平的变化,探讨ANGPTL3在大鼠肾组织中表达与蛋白尿的关系。方法通过激光微分离技术分离阿霉素肾病大鼠肾小球和肾小管,采用实时荧光定量 RT-PCR方法分析阿霉素注射后7、14、21和28 d肾组织中ANGPTL3 mRNA表达水平的变化。结果 (1)阿霉素注射后第14天开始出现大量蛋白尿(P<0．01),血清白蛋白明显减少 (P<0．01),胆固醇(P<0．01)和甘油三酯(P<0．05)明显增高,以上各项指标均在第28天达高峰。(2)通过激光微分离系统成功地分离了大鼠肾组织中的肾小球和肾小管。(3)阿霉素注射后7及14 d后,大鼠肾皮质中总ANGPTL3 mRNA表达水平与同一时间点正常对照组比较无明显变化;到第21天时为正常对照组的9．03倍(P<0．01);第28天时为正常对照组的 3．95倍(P<0．05),与第21天时相比已呈现下降趋势。(4)阿霉素注射后不同时间点肾小球中 ANGPTL3 mRNA表达水平的变化趋势与肾组织总ANGPTL3 mRNA表达水平的变化一致,第 7、14天时,肾小球中ANGPTL3 mRNA表达水平与同一时间点正常对照组比较无明显变化;到第21天时为同一时间点正常对照组的2．18倍(P<0．05);第28天时为正常对照组的1．90倍 (P<0．05)。(5)阿霉素注射后不同时间点肾小管中ANGPTL3 mRNA表达水平无明显改变,与同一时间点的正常对照组相比均无显著性差异。结论正常大鼠和阿霉素肾病大鼠的肾组织中均有ANGPTL3 mRNA表达。肾组织可以自身合成ANGPTL3;肾病的大鼠肾组织自身合成 ANGPTL3增多,且这种改变是发生在肾小球。ANGPTL3可能通过影响肾小球的功能参与肾病蛋白尿的进展过程。
Objective To observe the change of mRNA expression of angiopoietin-3 (ANGPTL3) in adriamycin-induced nephropathy rats during the development of proteinuria, and to explore the relationship between ANGPTL3 expression and proteinuria in rat kidney. Methods The glomeruli and tubules of rats with adriamycin-induced nephropathy were isolated by laser microdissection. The expression of ANGPTL3 mRNA in renal tissues at 7, 14, 21 and 28 days after doxorubicin injection was analyzed by real-time fluorescence quantitative RT-PCR Variety. Results (1) Massive proteinuria (P <0.01), serum albumin (P <0.01), cholesterol (P <0.01) and triglyceride (P <0.05), and the above indexes reached the peak on the 28th day. (2) Glomeruli and tubules in rat renal tissue were successfully separated by laser microdissection system. (3) After 7 and 14 days of doxorubicin injection, the expression level of total ANGPTL3 mRNA in rat renal cortex had no significant change compared with that of the normal control group at the same time point; it was 9.03 times (P < P <0.01). On the 28th day, it was 3.95 times that of the normal control group (P <0.05), which showed a decreasing trend compared with the 21st day. (4) The trend of ANGPTL3 mRNA expression in glomeruli at different time points after adriamycin injection was consistent with the change of total ANGPTL3 mRNA expression in renal tissue. On the 7th and 14th day, the expression level of ANGPTL3 mRNA in glomeruli was the same There was no significant difference between the control group and the normal control group at 21 days (P <0.05), and 1.90 times as much as the normal control group (P <0.05) at the 28th day .05). (5) The expression of ANGPTL3 mRNA in renal tubules did not change significantly at different time points after injection of doxorubicin, and no significant difference compared with the normal control group at the same time point. Conclusion ANGPTL3 mRNA expression in both normal and adriamycin-induced nephropathy rats. Renal tissue can synthesize ANGPTL3 by itself; nephropathy rat kidney tissue itself synthesis of ANGPTL3 increased, and this change occurs in the glomerular. ANGPTL3 may participate in the process of nephropathy proteinuria by affecting glomerular function.