金钱草提取物对糖尿病肾病大鼠肾脏保护作用及对肾组织Akt/GSK-3β信号通路的影响

Kidney Protection of Lysimachia Chinensis Extract on Rats with Diabetic Nephropathy and its Effect on Renal Akt/GSK-3β Signaling Pathway

目的:探讨金钱草提取物经Akt/GSK-3β信号通路对糖尿病肾病大鼠的肾脏保护作用。方法:SD大鼠120只,随机分为正常对照组、模型组、阳性对照组、金钱草提取物低、中、高剂量组,每组20只,雌雄各半。构建大鼠糖尿病肾病模型,并给予相应药物。检测大鼠血糖浓度和大鼠肾功能;制作HE切片,观察大鼠肾脏病理改变;检测大鼠肾脏Akt、GSK-3βmRNA表达水平;ELISA法检测大鼠肾脏Akt、GSK-3β蛋白水平表达。结果:正常对照组大鼠肾脏细胞结构完整,染色清晰,肾小管结构完整,模型组大鼠肾脏细胞大量坏死和肾小球出血;给予金钱草提取物和α-硫辛酸干预后,肾脏细胞结构完整,未见肾小球肥大及基底膜增厚。与正常对照组比较,模型组大鼠中期和末期血糖、24 h尿蛋白、肾脏指数、GSK-3βmRNA和蛋白表达水平均显著升高(P <0.05),肌酐清除率、Akt mRNA和蛋白表达水平显著降低(P <0.05)。给予金钱草提取物和α-硫辛酸干预后,金钱草提取物各剂量组和阳性对照组大鼠血糖、24 h尿蛋白、肾脏指数、GSK-3βmRNA和蛋白表达水平降低(P <0.05),且高剂量和中剂量组低于低剂量组,高剂量组低于中剂量组(P <0.05),阳性对照组和高剂量组差异无统计学意义(P> 0.05);肌酐清除率、Akt mRNA和蛋白表达水平升高(P <0.05),且高剂量和中剂量组高于低剂量组,高剂量组高于中剂量组(P <0.05),阳性对照组和高剂量组差异无统计学意义(P> 0.05)。结论:金钱草提取物对糖尿病大鼠的肾脏具有保护作用,其机制可能是通过抑制GSK-3β、促进Akt蛋白的表达水平,从而扭转糖尿病对大鼠肾脏造成的损伤。

Objective: To investigate the Kidney protection of Lysimachia chinensis extract on rats with diabetic nephropathy and its effect on renal Akt/GSK-3β signaling pathway. Methods: 120 SD rats were randomly divided into normal control group,model group,positive control group,low,medium and high dose groups of Lysimachia chinensis extracts with 20 rats in each group,half male and half female. A rat model of diabetic nephropathy was constructed and the corresponding drugs were administered. The blood glucose concentration and renal function of rats were detected. HE sections were made and the pathological changes of rat kidney were observed. The expressions of Akt and GSK-3β mRNA in rat kidney were detected. The expression of Akt and GSK-3β protein in rat kidney was detected by ELISA. Results: In the normal control group,the structure of renal cells was complete,the staining was clear,and the structure of renal tubules was complete. In the model group,a large number of renal cells were necrotic and glomerular hemorrhage. After the treatment with lysimachia chinensis extract and α-lipoic acid,the structure of renal cells was complete without glomerular hypertrophy and basement membrane thickening. Compared with the normal control group,the blood glucose,24 h urine protein,kidney index,GSK-3β mRNA and protein expression level of the model group were significantly increased(P < 0. 05),and the creatinine clearance rate,Akt mRNA and protein expression level were significantly decreased(P < 0. 05). After the treatment of lysimachia chinensis extract and α-lipoic acid,the blood glucose and 24 h urine Protein,kidney index,GSK-3β mRNA and protein expression levels of lysimachia chinensis extract each dose group and the positive control group were decreased(P <0. 05),in which those in the high dose and middle dose groups were lower than those in the low dose group,those in the high dose group was lower than those in the middle dose group(P < 0. 05),and there was no difference between the positive control group and the high dose group(P > 0. 05);the creatinine clearance rate,Akt mRNA and protein expression level were increased(P < 0. 05),in which those in the high dose and middle dose groups were higher than those in the low dose group,and those in the high dose group higher than those in the middle dose group. There was no difference between positive control group and high dose group(P > 0. 05).Conclusion: The extract of Lysimachia chinensis has a protective effect on the kidney of diabetic rats. The mechanism may be to inhibit the damage of the kidney of rats by inhibiting GSK-3β and promoting the expression level of Akt protein.