目的:评价腺苷酸活化蛋白激酶(AMPK)在小檗碱减轻肾缺血再灌注小鼠肾纤维化中的作用及其与铁死亡的关系。方法:SPF级健康成年雄性C57BL/6小鼠24只，8～10周龄，体重20～25 g，采用随机数字表法分为4组(n n＝6)：假手术组(S组)、肾缺血再灌注组(I/R组)、小檗碱组(B组)和小檗碱＋AMPK抑制剂Compound C组(BC组)。采用夹闭左肾肾蒂45 min后恢复灌注的方法制备小鼠肾缺血再灌注模型。B组和BC组术前14 d连续进行小檗碱100 mg/kg灌胃，BC组术前3 d连续腹腔注射Compound C 0.2 mg/kg，S组和I/R组给予等容量生理盐水，于再灌注第14天眶静脉采集血标本，检测血清Cr和BUN浓度，随后处死小鼠取肾组织，检测Fen 2＋、MDA含量和SOD活性，ELISA法检测IL-β和TNF-α含量，光镜下观察病理学结果，Western blot法检测p-AMPKα、长链脂酰辅酶A合成酶4(ACSL4)和谷胱甘肽过氧化物酶(GPX4)的表达。透射电镜下观察细胞线粒体超微结构。n 结果:与S组比较，I/R组、B组和BC组血清Cr和BUN浓度、肾小管损伤评分和肾纤维化评分升高，I/R组和BC组肾组织MDA、IL-1β和TNF-α含量升高，SOD活性降低，I/R组和B组肾组织p-AMPKα和ACSL4、BC组ACSL4、B组GPX4表达上调，I/R组和BC组GPX4表达下调，I/R组和BC组Fen 2＋含量升高(n P<0.05)；与I/R组比较，B组和BC组血清Cr和BUN浓度、肾小管损伤评分和肾纤维化评分降低，B组肾组织MDA、IL-1β和TNF-α含量降低，SOD活性升高，B组肾组织p-AMPKα和GPX4、BC组ACSL4表达上调，BC组肾组织p-AMPKα和GPX4、B组ACSL4表达下调(n P<0.05)；与B组比较，BC组血清Cr和BUN浓度、肾小管损伤评分和肾纤维化评分升高，MDA、IL-1β和TNF-α含量升高，SOD活性降低，p-AMPKα和GPX4表达下调，ACSL4表达上调，Fen 2＋含量升高(n P<0.05)。BC组肾组织病理学损伤和超微结构损伤较B组加重。n 结论:小檗碱可明显减轻肾缺血再灌注小鼠肾纤维化，其机制与AMPK激活进而抑制铁死亡有关。“,”Objective:To evaluate the role of Adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) in berberine-induced reduction of renal fibrosis in a mouse model of renal ischemia-reperfusion (I/R) and the relationship with ferroptosis.Methods:Twenty-four SPF healthy adult male C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups (n n=6 each) by a random number table method: sham operation group (group S), renal I/R group (I/R group), berberine group (B group), and berberine plus AMPK inhibitor Compound C group (BC group). The renal I/R model was established by clamping the renal pedicle of the left kidney for 45 min followed by reperfusion in anesthetized mice.In B and BC groups, berberine 100 mg/kg was given for continuous intragastric administration at 14 days before surgery, Compound C 0.2 mg/kg was intraperitoneally injected continuously at 3 days before surgery in group BC, and the equal volume of normal saline was given instead in S and I/R groups.Blood samples were collected from the orbital vein on the 14th day of reperfusion to measure serum Cr and BUN concentrations.Then the mice were sacrificed and kidney tissues were removed for determination of the Fen 2+ and malondialdehyde (MDA) contents and superoxide dismutase (SOD) activity, interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents (by enzyme-linked immunosorbent assay), and expression of phosphorylated AMPKα (p-AMPKα), Acyl-CoA synthetase long-chain familymember4 (ACSL4) and recombinant glutathione peroxidase 4 (GPX4) (by Western blot) and for examination of the pathological changes (with a light microscope) and ultrastructure of cell mitochondria (with a transmission electron microscope). The damage to the renal tubules was scored.n Results:Compared with group S, serum Cr and BUN concentrations, renal tubular damage score and renal fibrosis score were significantly increased in I/R, B and BC groups, the contents of MDA, IL-1β and TNF-α were significantly increased, and SOD activity was decreased in I/R and BC groups, the expression of p-AMPKα and ACSL4 in I/R and BC groups, ACSL4 in BC group and GPX4 in B group was significantly up-regulated, and the expression of GPX4 was significantly down-regulated, and the Fen 2+ content was increased in I/R and BC groups (n P<0.05). Compared with I/R group, serum Cr and BUN concentrations, renal tubular damage score and renal fibrosis score were significantly decreased in B and BC groups, the contents of MDA, IL-1β and TNF-α were significantly decreased, and the activity of SOD was increased in group B, the expression of p-AMPKα and GPX4 in B group and ACSL4 in BC group was up-regulated, and the expression of p-AMPKα and GPX4 in BC group and ACSL4 in B group was down-regulated (n P<0.05). Compared with group B, serum Cr and BUN concentrations, renal tubular damage score and renal fibrosis score were increased, the contents of MDA, IL-1β and TNF-α were increased, SOD activity was decreased, the expression of p-AMPKα and GPX4 was down-regulated, the expression of ACSL4 was up-regulated, and the content of Fen 2+ was increased in BC group (n P<0.05). The pathological and ultrastructural damage of kidney tissues was more serious in group BC than in group B.n Conclusion:Berberine can significantly reduce renal fibrosis in a mouse model of renal I/R, and the mechanism is related to the activation of AMPK and further inhibition of ferroptosis.