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目的:研究中药复方健脾清化方对慢性肾衰竭肾纤维化大鼠ATⅡ/NADPH氧化应激通路的影响,初步探讨其发挥疗效的作用机制。方法:用5/6肾切除(Plat法)建立慢性肾衰竭大鼠模型,分为假手术组、模型组、健脾清化方组和氯沙坦组。治疗60 d后测定血清肌酐、尿素氮水平;测定各组肾组织中SOD和MDA水平;Western印迹检测各组肾组织AT1蛋白表达;RT-PCR检测各组肾组织p47phox mRNA表达。结果:与假手术组比较,模型组大鼠血清肌酐和尿素氮均有明显上升,肾组织匀浆中SOD活性降低,MDA含量升高,肾组织中AT1蛋白表达显著上调,p47phox mRNA表达显著增加,差异均具有统计学意义(P<0.05)。与模型组比较,健脾清化方组和氯沙坦组血清肌酐和尿素氮含量降低,肾组织匀浆中SOD活性上升,MDA含量下降,肾组织中AT1蛋白表达显著下调,p47phox mRNA表达显著降低,差异均具有统计学意义(P<0.05)。与氯沙坦组比较,健脾清化方组肾组织SOD活性明显升高,(P<0.05),肾组织中AT1蛋白及p47phox mRNA表达下降趋势明显,但差异无统计学意义(P>0.05),血清BUN,SCr水平和肾组织中MDA含量两组间比较差异也无统计学意义(P>0.05)。结论:健脾清化方可通过降低ATⅡ及NADPH氧化酶的表达,从而改善慢性肾衰竭大鼠的氧化应激反应,延缓肾纤维化进程。
Objective: To investigate the effect of Jianpi Qinghua Recipe on AT Ⅱ / NADPH oxidative stress pathways in rats with renal failure induced by chronic renal failure and to explore its mechanism of action. Methods: Chronic renal failure rat model was established by 5/6 nephrectomy (Plat method), and divided into sham operation group, model group, Jianpi Qinghua Recipe group and losartan group. Serum creatinine and urea nitrogen levels were measured after 60 days of treatment. The levels of SOD and MDA in renal tissue of each group were determined. The expression of AT1 protein in renal tissues was detected by Western blotting. The expression of p47phox mRNA in renal tissues was detected by RT-PCR. Results: Compared with the sham-operation group, serum creatinine and urea nitrogen in model group were significantly increased, SOD activity in renal homogenate was decreased, MDA content was increased, AT1 protein expression in renal tissue was significantly increased, p47phox mRNA expression was significantly increased , The differences were statistically significant (P <0.05). Compared with the model group, serum creatinine and urea nitrogen in the Jianpi Qinghua Recipe group and the losartan group decreased, SOD activity in renal tissue homogenate increased, MDA content decreased, AT1 protein expression in renal tissue was significantly decreased, p47phox mRNA expression was significantly Decreased, the differences were statistically significant (P <0.05). Compared with Losartan group, the activity of SOD in renal tissue of Jianpi Qinghua Recipe group was significantly increased (P <0.05), while the expression of AT1 protein and p47phox mRNA in renal tissue decreased significantly, but the difference was not statistically significant (P> 0.05 ), Serum BUN, SCr levels and renal tissue MDA content was no significant difference between the two groups (P> 0.05). Conclusion: Jianpi Qinghua Decoction can reduce oxidative stress of ATⅡand NADPH oxidase and improve the oxidative stress response in rats with chronic renal failure and delay the process of renal fibrosis.