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目的通过观察染铅大鼠肾脏纤维化相关基因核因子κB(NFκB)、转化生长因子β(TGFβ)和纤维连接蛋白(FN)的表达,探讨铅导致大鼠肾脏纤维化的分子机制。方法将32只大鼠分为4组:对照组、1个月染铅组、2个月染铅组、3个月染铅组,分别以蒸馏水、质量分数为0.5%的醋酸铅喂饮。应用免疫组化的方法检测肾皮质NFκB、TGFβ及FN的表达情况,并用逆转录-聚合酶链式反应(RTPCR)的方法检测肾皮质中上述指标mRNA的表达。结果染铅组NFκB蛋白的表达增加(1、2、3个月染铅组分别为0.2315±0.0624、0.3213±0.0740、0.4729±0.0839),与对照组(0.1464±0.0624)相比,差异有统计学意义(P<0.05);3组染铅组NFκBmRNA的表达也明显增加,其与相应内参照的灰度比分别为0.4370±0.0841、0.5465±0.0503、0.6443±0.0538,与对照组(0.3608±0.0550)相比,差异有统计学意义(P<0.05)。3个月染铅组TGFβmRNA表达(0.5225±0.0416)明显增强,与对照组的差异有统计学意义(P<0.05),其余组与对照组(0.4645±0.0461)相比,差异无统计学意义(P>0.05),免疫组化结果显示,TGFβ在各组中的差异无统计学意义(P<0.05)。FN蛋白的表达在2个月组(0.4243±0.0595)和3个月组(0.4917±0.0891)与对照组(0.3530±0.0490)比较,差异有统计学意义(P<0.05);其mRNA的表达同样在2个月组(0.8650±0.0880)和3个月组(0.8714±0.0980)与对照组(0.7432±0.0639)的差异有统计学意义(P<0.05)。结论染铅大鼠肾脏NFκB、TGFβ及FN表达增加可能为铅所导致肾脏纤维化的重要环节。
OBJECTIVE: To investigate the molecular mechanism of lead-induced renal fibrosis in rats by observing the expression of nuclear factor-kappa B (NFκB), transforming growth factor β (TGFβ) and fibronectin (FN) in kidney of lead-exposed rats. Methods 32 rats were divided into 4 groups: the control group, the lead group for 1 month, the lead group for 2 months and the lead group for 3 months. The rats were fed with distilled water and 0.5% lead acetate respectively. The expression of NFκB, TGFβ and FN in renal cortex was detected by immunohistochemistry. The mRNA expression of these indexes in renal cortex was detected by reverse transcriptase-polymerase chain reaction (RTPCR). Results The expression of NFκB was increased in lead-exposed group (0.2315 ± 0.0624,0.3213 ± 0.0740,0.4729 ± 0.0839, respectively) in the lead-exposed group (P <0.05), which was significantly lower than that in the control group (0.1464 ± 0.0624) (P <0.05). The expression of NFκBmRNA in the three lead-exposed groups also increased significantly (P <0.05). The grayscale ratios were 0.4370 ± 0.0841,0.5465 ± 0.0503,0.6443 ± 0.0538 with the control group (0.3608 ± 0.0550) Compared with the difference was statistically significant (P <0.05). The expression of TGFβmRNA in the lead-exposed group at 3 months (0.5225 ± 0.0416) was significantly increased compared with the control group (P <0.05), but there was no significant difference between the other groups and the control group (0.4645 ± 0.0461) P> 0.05). The results of immunohistochemistry showed that there was no significant difference of TGFβ in each group (P <0.05). The expression of FN protein in two-month group (0.4243 ± 0.0595) and three-month group (0.4917 ± 0.0891) and control group (0.3530 ± 0.0490), the difference was statistically significant (P <0.05) The differences between the two groups (0.8650 ± 0.0880) and the 3 months group (0.8714 ± 0.0980) and the control group (0.7432 ± 0.0639) were statistically significant (P <0.05). Conclusion The increased expression of NFκB, TGFβ and FN in lead-exposed rat kidney may be an important link of lead-induced renal fibrosis.