GSK-3β抑制剂氯化锂对糖尿病肾病大鼠肾组织核因子κB受体活化因子与其配体表达的影响

Effect of GSK-3β inhibitor on the expression of RANK-RANKL in rats kidney tissue with diabetic nephropathy

目的 探讨GSK-3β抑制剂氯化锂及核因子κB受体活化因子(RANK)与核因子κB受体活化因子配体(RANKL)对糖尿病肾病(DN)大鼠肾组织病理改变的影响及意义.方法 将实验大鼠分成正常对照组(NC组)、DN模型组(DN组)和GSK-3β抑制剂氯化锂干预组( LiCl组),各16只.考马斯亮蓝法检测24 h尿蛋白;肾脏组织切片行HE染色,观察大鼠肾脏病理改变;即时荧光定量逆转录聚合酶链反应(RT-qPCR)及免疫组织化学法(IHC)检测各组肾组织GSK-3β、RANK、RANKL的mRNA及蛋白的表达.结果 与NC组(14. 72±3. 37)g/24 h相比,DN组24 h尿蛋白水平(154. 17± 20. 65)g/24 h升高;与DN组相比,LiCl组24 h尿蛋白水平(107. 22±31. 15)g/24 h下降(P<0. 05).与NC组[(2. 10±0. 60)、(1. 10±0. 20)、(1. 21±0. 20);(19. 52±3. 20)、(1. 80±1. 10)、(1. 81±0. 50)]相比,DN组大鼠肾组织病理改变加重,GSK-3β、RANK、RANKL的mRNA及蛋白表达[(9. 10±2. 15)、(8. 95±2. 40)、(9. 90±2. 60);(32. 70±7. 20)、(19. 20±4. 32)、(20. 92±5. 90)]升高;与DN组相比,LiCl组大鼠肾组织病理改变减轻,GSK-3β、RANK、RANKL的mRNA及蛋白表达[(2. 70±0. 80)、(2. 32± 0. 65)、(3. 58±1. 10);(22. 35±3. 25)、(4. 20±2. 42)、(5. 90±2. 36);均P<0. 05]下降.结论 RANK和RANKL在DN发生发展中有重要作用;LiCl可通过激活Wnt信号通路,抑制GSK-3β,下调RANK和RANKL的表达,可一定程度上改善DN中的肾脏损害.

Objective To investigate the effect and significance of GSK-3β inhibitor (LiCl)and RANK-RANKL on the renal tissue of diabetic nephropathy(DN)rats.Methods SD rats were divided into normal control group (NC),DN model group (DN)and GSK-3β inhibitor intervention group (LiCl). Twenty-four hour urine protein of rats were determined by Coomassie brilliant blue.Kidney tissue sections were stained by HE.The expression of GSK-3β,RANK and RANKL protein were determined by immunohistochemistry staining.The mRNA of GSK-3β,RANK,RANKL was detected by RT-qPCR.Results Compared with NC group[(14.72±3.37)g ],the level of 24-hour urinary protein [(154.17±20.65)g ] increased significantly in DN group;compared with DN Group,the level of 24-hour urinary protein [(107.22±31.15)g]decreased in LiClgroup(P<0.05).Compared with NC group (2.10±0.60,1.10± 0.20,1.21±0.20;19.52±3.20,1.80±1.10,1.81±0.50),the pathological changes of renal tissues of DN group aggravated,the mRNA and expression of protein of GSK-3β,RANK and RANKL increased(9.10± 2.15,8.95±2.40,9.90±2.60;32.70±7.20,19.20±4.32,20.92±5.90);compared with DN group,the pathological changes of renal tissues of LiClgroup alleviated,mRNA and the expression of protein of factors above declined(2.70±0.80,2.32±0.65,3.58±1.10;22.35±3.25,4.20±2.42,5.90±2.36;P<0.05). Conclusion RANK and RANKL play an important role in the development of DN,LiClinfluence Wnt and NF-κB signal pathway down-regulating RANK and RANKL to suspend development of diabetic nephropathy.