【摘要】 目的探讨黑果枸杞(LyciumruthenicumMurray)汁调控核因子E2相关因子2(nuclearfactorerythroid2-relatedfactor2,Nrf2)信号通路蛋白质表达改善大鼠酒精性肾损伤的作用机制。方法 60只6w龄雄性SD大鼠随机分为空白对照组(C)、模型组(M)、低剂量黑果枸杞汁+模型组(LLM)、中剂量黑果枸杞汁+模型组(MLM)、高剂量黑果枸杞汁+模型组(HLM),每组12只。M、LLM组、MLM、HLM组每天13:00、19:00以20ml/kg [8 g/(kg bw·d)]分2次灌胃400 g/L乙醇,C组于相同时间点施以等体积蒸馏水。黑果枸杞汁各剂量组酒精灌胃4h前,分别以2.4、4.8、9.6 ml/(bw·d)黑果枸杞汁进行灌胃,其他组于相当时间点分别施以等体积蒸馏水。实验周期4w,末次实验结束后24h处死大鼠,取血及肾脏,计算肾脏指数,HE染色观察肾脏组织形态,免疫组化法检测肾脏Nrf2及磷酸化Nrf2(phosphorylated nuclear factor erythroid 2-related factor 2,p-Nrf2)、血红素氧合酶(heme oxygenase-1,HO-1)蛋白质表达,Jaffe苦味酸法检测血清肌酐(serum creatinine,Scr),二乙酰-肟法检测血清尿素氮(blood urea nitrogen,BUN),比色法测定大鼠肾脏组织超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malonaldehyde,MDA),荧光光度法测定活性氧簇(reactive oxygen species,ROS)。结果与C组比较,M组肾脏组织形态发生病理改变;肾脏指数、Scr、BUN、肾脏MDA和ROS含量显著升高(P<0.05或P<0.01);肾脏SOD活性、p-Nrf2和HO-1蛋白质表达、p-Nrf2/Nrf2显著降低(P<0.01)。与M组比较,LLM、MLM、HLM组肾脏组织形态均有所改善,其中以HLM组最为显著;肾脏指数,LLM、MLM和HLM组均显著降低(P<0.05或P<0.01);BUN和Scr含量,MLM和HLM组均显著降低(P<0.05或P<0.01);肾脏p-Nrf2和HO-1蛋白质表达,MLM和HLM组均显著升高(P<0.05或P<0.01);肾脏p-Nrf2/Nrf2,HLM组显著升高(P<0.01);肾脏SOD活性,LLM、MLM和HLM组均显著升高(P<0.05或P<0.01);肾脏MDA含量,LLM、MLM和HLM组均显著降低(P<0.05或P<0.01);肾脏ROS含量,MLM和HLM组均显著降低(P<0.01)。黑果枸杞汁各剂量组组间比较,BUN含量,LLM和MLM组均显著高于HLM组(P<0.01);Scr含量,LLM组显著高于HLM组(P<0.05);肾脏p-Nrf2蛋白质表达,LLM和MLM组均显著低于HLM组(P<0.05或P<0.01);肾脏p-Nrf2/Nrf2和HO-1蛋白质表达,LLM组显著低于HLM组(P<0.05);肾脏SOD活性,LLM和MLM组均显著低于HLM组(P<0.05或P<0.01);肾脏MDA含量,LLM和MLM组均显著高于HLM组(P<0.01);肾脏ROS含量,LLM组显著高于HLM组(P<0.01)。结论黑果枸杞汁可以通过调控Nrf2信号通路,改善氧化应激,改善大鼠酒精性肾损伤。[营养学报,2021,43(5):485-491]更多还原

【Abstract】 Objective To explore the mechanism of the protection of Lycium ruthenicum Murray juice against alcoholic kidney injury by regulating the protein expression of nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway in rats. Methods Sixty 6-week-old male SD rats were randomly divided into control group(C), model group(M),low-dose Lycium ruthenicum Murray juice + model group(LLM), medium-dose Lycium ruthenicum Murray juice+model group(MLM) and high-dose Lycium ruthenicum Murray juice + model group(HLM)(12 rats per group). Rats in M, LLM,MLM and HLM groups were given 20 ml/kg(8 g/kg bw·d) ethanol(400 g/L) intragastrically at 13:00, 19:00 respectively, and rats in group C were given an equal volume of distilled water at the same time points. Four hours prior to alcohol gavage, 2.4,4.8, 9.6 ml/(bw·d) Lycium ruthenicum Murray juice were administered in corresponding dose groups, respectively, and the other groups were given equal volume of distilled water at the same time points. The intervention period lasted four weeks.Then, rats were sacrificed 24 hours after the end of the last treatment, blood and kidney were collected. The kidney was weighed to calculate kidney index. The morphology of the kidney was observed by HE staining. The expression of Nrf2,phosphorylated nuclear factor erythroid 2-related factor 2(p-Nrf2) and heme oxygenase-1(HO-1) in kidney were detected by immunohistochemistry. The serum creatinine(Scr) was detected by Jaffe’s method(picric acid). The blood urea nitrogen(BUN) was detected by diacetyl-oxime method. The renal superoxide dismutase(SOD) and malondialdehyde(MDA) were detected by colorimetry. The renal reactive oxygen species(ROS) was detected by fluorescence spectrometry. Results Compared with group C, histopathological changes were observed in group M. The kidney index, SCr, BUN, renal MDA and ROS concentrations increased significantly(P<0.05 or P<0.01). Renal SOD activity, expression of p-Nrf2 and HO-1,p-Nrf2/Nrf2 decreased significantly(P<0.01). Compared with group M, the morphology of kidney was improved in group LLM, MLM and HLM, and the improvement was the most in group HLM. The kidney index, decreased significantly in group LLM, MLM and HLM(P<0.05 or P<0.01). BUN and Scr concentration decreased significantly in group MLM and HLM(P<0.05 or P<0.01). Renal p-Nrf2 and HO-1 expression increased significantly in group MLM and HLM(P<0.05 or P<0.01).Renal p-Nrf2/Nrf2, increased significantly in group HLM(P<0.01). Renal SOD activity increased significantly in group LLM,MLM and HLM(P<0.05 or P<0.01). Renal MDA concentration decreased significantly in group LLM, MLM and HLM(P<0.05 or P<0.01). Renal ROS concentration decreased significantly in group MLM and HLM(P<0.01). Comparison among the dose groups of Lycium ruthenicum Murray juice, BUN concentration were significantly higher in both group LLM and MLM than in group HLM(P<0.01). Scr concentration was significantly higher in group LLM than in group HLM(P<0.05).Renal p-Nrf2 expression were significantly lower in both group LLM and MLM than in group HLM(P<0.05 or P<0.01).Renal p-Nrf2/Nrf2 and HO-1 expression was significantly lower in group LLM than in group HLM(P<0.05). Renal SOD activity were significantly lower in both group LLM and MLM than in group HLM(P<0.05 or P<0.01). Renal MDA concentration were significantly higher in both group LLM and MLM than in group HLM(P<0.01). Renal ROS concentration was significantly lower in group LLM than in group HLM(P<0.01). Conclusion Lycium ruthenicum Murray juice can prevent or reduce oxidative stress and attenuate alcoholic kidney injury in rats by regulating the Nrf2 signaling pathway. [ACTA NUTRIMENTA SINICA, 2021,43(5):485-491]更多还原