盐酸戊乙奎醚预先给药对横纹肌溶解致急性肾损伤大鼠肾组织Nrf2／HO-1信号通路的影响

Effect of penehyclidine hydroch｜oride pretreatment on Nrf2/HO-1 signaling pathway in renal tissues of rats with rhabdomyolysis-induced acute kidney injury

目的探讨盐酸戊乙奎醚预先给药对横纹肌溶解致急性肾损伤大鼠肾组织核因子E2相关因子2/血红素加氧酶-1（Nrf2/HO-1）信号通路的影响。 方法清洁级雄性SD大鼠36只，体重200～220 g，采用随机数字表法分为3组（n＝12）：对照组（C组）、急性肾损伤组（AKI组）和盐酸戊乙奎醚预先给药组（PHC组）。采用双侧后肢肌肉注射50%甘油生理盐水溶液10 ml/kg的方法制备横纹肌溶解症模型。PHC组于肌肉注射甘油前30 min时腹腔注射盐酸戊乙奎醚0.2 mg/kg。于肌肉注射甘油后1和6 h时采集血样，测定血清BUN和Cr浓度；取双肾，行HE染色，观察病理学结果，行肾小管损伤评分；采用底物反应法测定肾组织HO-1活性，采用RT-qPCR法测定Nrf2和HO-1的mRNA表达水平，采用Western blot法测定肾组织核蛋白和总蛋白Nrf2和总蛋白HO-1的表达水平。 结果与C组比较，AKI组和PHC组血清BUN和Cr浓度和肾小管损伤评分升高，肾组织核蛋白和总蛋白Nrf2、总蛋白HO-1表达上调，HO-1活性增强，AKI组肾组织HO-1 mRNA表达上调，PHC组肾组织Nrf2 mRNA和HO-1 mRNA表达上调（P〈0.01或0.05）；与AKI组比较，PHC组血清BUN和Cr浓度和肾小管损伤评分降低，肾组织Nrf2 mRNA和HO-1 mRNA、核蛋白和总蛋白Nrf2、总蛋白HO-1表达上调，HO-1活性增强（P〈0.01或0.05）。 结论盐酸戊乙奎醚预先给药减轻大鼠横纹肌溶解致急性肾损伤的机制可能与激活肾组织Nrf2/HO-1信号通路有关。

Objective To investigate the effect of penehyclidine hydrochloride （PHC） pretreat- ment on nuclear faetor erythroid 2-related factor 2/heine oxygenase-1 （Nrf2/HO-1） signaling pathway in re- nal tissues of rats with rhabdomyolysis-induced acute kidney injury （ AKI）. Methods Thirty-six pathogen- free male Sprague-Dawley rats, weighing 200-220 g, were assigned into 3 groups （n = 12 each） using a random number table： control group （group C） , group AKI and PHC pretreatment group （group PHC）. Rhabdomyolysis was induced by intramuscular injection of 50% glycerol 10 ml/kg in bilateral hindlimbs. PHC 0.2 mg/kg was injected intraperitoneally at 30 min before glycerol was injected intramuscularly in group PHC. At 1 and 6 h after glycerol injection, serum was collected for determination of blood urea nitrogen （BUN） and creatinine （Cr） concentrations, and bilateral kidneys were harvested for pathological ex- amination and for determination of HO-1 activity and expression of Nrf2 mRNA and HO-1 mRNA （by quan- titative real-time polymerase chain reaction） , Nrf2 in nucleoprotein and total protein and HO-1 in total pro- tein in renal tissues （by Western blot）. The damage to the renal tubules was scored. Results Compared with group C, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly increased, the expression of Nrf2 in nucleoprotein and total protein and HO-1 in total protein was signifi- cantly up-regulated, and HO-1 activity was significantly increased in AK1 and PHC groups, the expression of HO-1 mRNA was significantly up-regulated in group AK1, and the expression of Nrf2 mRNA and HO-I mRNA was significantly up-regulated in group PHC （P〈0.01 or 0.05）. Compared with group AKI, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly decreased, the ex- pression of Nrf2 in nucleoprotein and total protein and HO-I in total protein was significantly up-regulated, and HO-1 activity was significantly increased in group PHC （P〈0.01 or 0. 05）. Conclusion The mechanism by which PHC pretreatment attenuates rhabdomyolysis-induced AK1 may be related to activation of Nrf2/HO-1 signaling pathway in renal tissues of rats.