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目的探讨幼年大鼠持续蛋白尿致肾损伤的不同时间点肾组织NF-κB亚单位P65/Rel-A及凝血酶敏感蛋白(TSP-1)、转化生长因子(TGF-β1)、结缔组织生长因子(CTGF)和纤维连接蛋白(FN)mRNA动态表达的趋势。方法3周龄幼年W istar雌性大鼠80只,分为BSA组(40只)和对照组(40只),采用BSA诱导制备蛋白负荷肾病模型;考马斯亮蓝比色测大鼠尿蛋白;HE染色评价肾组织常病理变化;原位杂交检测P65/Rel-A、TSP-1、TGF-β1及CTGF mRNA表达;Northern b lot检测FNmRNA表达。SPSS10对实验数据进行统计学处理。结果(1)BSA组大鼠至3~4周蛋白尿达大量蛋白尿程度,肾间质炎症细胞浸润,肾小管蛋白管型形成,间质水肿,间质区加宽。(2)BSA组各级肾小管上皮细胞P65/Rel-A mRNA于细胞核的表达强度趋于增加,第1、2、3、4周半定量积分分别为:2.33±0.20、2.76±0.12、2.96±0.19、3.76±0.18(F=37.34,P<0.01)。(3)BSA组TSP-1 mRNA表达第1、2、3、4周半定量积分分别为:2.60±0.28、3.39±0.41、2.77±0.08、2.71±0.13,于第2周时达高峰(F=11.14,P<0.01)。(4)伴随蛋白尿的加重,BSA组TGF-β1及CTGF的mRNA于各级肾小管上皮细胞表达趋势进行性增强,与对照组比较及自身各时间点比较差异均有统计学意义(P<0.01)。(5)Northern b lot结果提示,BSA组FN mRNA于第2周明显上调,至第4周是对照组的3.6倍,是第1周的2.7倍,与对照组比较差异均有统计学意义(P<0.01)。结论大量持续蛋白尿过程中肾组织NF-κB信号途径活化,TSP-1、TGF-β1、CTGF的异常表达,肾间质FN的合成和异常集聚,可能是促进肾间质进一步损伤的机制之一。
Objective To investigate the expressions of P65 / Rel-A and TSP-1, TGF-β1 and connective tissue growth factor (NF-κB) in kidneys at different time points after continuous proteinuria- (CTGF) and fibronectin (FN) mRNA trends in the dynamic expression. Methods 80 Wistar female rats aged 3 weeks were divided into BSA group (40) and control group (40). BSA was used to induce the protein load nephropathy model. Coomassie blue colorimetry The expression of P65 / Rel-A, TSP-1, TGF-β1 and CTGF mRNA were detected by in situ hybridization. The expression of FN mRNA was detected by Northern b lot. SPSS10 experimental data for statistical analysis. Results (1) In the BSA group, proteinuria reached a large amount of proteinuria by 3-4 weeks, infiltration of interstitial inflammatory cells, tubular protein tubular formation, interstitial edema and widening of interstitial region. (2) The expression intensity of P65 / Rel-A mRNA in nuclei of tubular epithelial cells at all levels in BSA group tended to increase. The integrals at the first, second, third and fourth week were 2.33 ± 0.20, 2.76 ± 0.12 and 2.96 respectively ± 0.19, 3.76 ± 0.18 (F = 37.34, P <0.01). (3) The semi-quantitative integral of TSP-1 mRNA expression at 1, 2, 3 and 4 weeks in BSA group was 2.60 ± 0.28, 3.39 ± 0.41, 2.77 ± 0.08 and 2.71 ± 0.13 respectively, reaching the peak at the second week (F = 11.14 , P <0.01). (4) With the exacerbation of proteinuria, the expression of TGF-β1 and CTGF mRNA in BSA group increased progressively at all levels of renal tubular epithelial cells, compared with the control group and their own time points were statistically significant (P < 0.01). (5) The result of Northern b lot indicated that the FN mRNA in BSA group was significantly up-regulated in the second week, which was 3.6 times of the control group in the fourth week and 2.7 times that in the first week, which was significantly different from the control group P <0.01). Conclusion Abnormal expression of TSP-1, TGF-β1 and CTGF, synthesis and abnormal aggregation of renal interstitial FN may be the mechanism of further damage of renal interstitium in a large number of persistent proteinuria one.