Alpha-cyperone ameliorates renal fibrosis and inflammation in mice with chronic kidney disease <i>via</i> NF-κB and Akt/Nrf2/HO-1 pathways

Renal fibrosis is a hallmark feature of chronic kidney disease (CKD) and contributes to local inflammation and impaired renal function. The study aimed to investigate the effects of an active compound, alpha-cyperone, on renal fibrosis, inflammation, and oxidative stress in CKD mice. After subtotal nephrectomy, mice were administered with alpha-cyperone (5, 10, and 20 mg/kg) or captopril (the positive control) daily <i>via</i> oral gavage. Masson trichrome staining and periodic Acid-Schiff staining were performed to measure fibrotic areas and renal histopathological changes. An enzyme-linked immunosorbent assay was conducted to measure inflammatory cytokine levels in mouse serum samples. Superoxide anion concentrations, reactive oxygen species (ROS) level, and superoxide dismutase (SOD) activity were analyzed. Immunohistochemistry was performed to detect inflammatory and oxidative stress markers (Ly6G and 8-OHdG) in mouse renal tissues. Protein levels of factors involved in NF-κB signaling and Akt/Nrf2/HO-1 signaling were quantified by western blotting. Alpha-cyperone at the dose of 10 mg/kg significantly improved renal functions in CKD model mice and alleviated tubulointerstitial fibrosis, with effects comparable to those of captopril. After CKD modeling, serum levels of inflammatory mediators, Ly6G levels, superoxide anion activity, ROS levels, and 8-OHdG levels were markedly increased, while the activity of antioxidant enzyme SOD was reduced. All these changes were ameliorated by alpha-cyperone or captopril treatment. Moreover, alpha-cyperone inhibited the NF-κB signaling and activated the Akt/Nrf2/HO-1 signaling in renal tissues of CKD mice. Alpha-cyperone ameliorates renal fibrosis in CKD mice by inhibiting inflammation and oxidative stress <i>via</i> NF-κB and Akt/Nrf2/HO-1 pathways.肾纤维化是慢性肾脏病（chronic kidney disease, CKD）的标志性病理特征，可介导局部炎症反应并造成肾功能损伤。本研究旨在探究活性化合物α-香附酮（alpha-cyperone）对慢性肾脏病模型小鼠肾纤维化、炎症反应及氧化应激的影响。小鼠经次全肾切除术（subtotal nephrectomy）构建慢性肾脏病模型后，每日分别以α-香附酮（剂量为5、10、20 mg/kg）或卡托普利（captopril，阳性对照药物）经口灌胃给药。采用马松三色染色（Masson trichrome staining）与过碘酸-雪夫染色（periodic Acid-Schiff staining）评估纤维化面积及肾脏组织病理形态学变化；通过酶联免疫吸附试验（enzyme-linked immunosorbent assay）检测小鼠血清炎症细胞因子水平；测定超氧阴离子（superoxide anion）浓度、活性氧（reactive oxygen species, ROS）水平及超氧化物歧化酶（superoxide dismutase, SOD）活性；采用免疫组化法（immunohistochemistry）检测小鼠肾组织中炎症与氧化应激标志物Ly6G及8-羟基脱氧鸟苷（8-OHdG）的表达情况；通过蛋白质免疫印迹法（western blotting）定量分析NF-κB信号通路（NF-κB signaling）与Akt/Nrf2/HO-1信号通路（Akt/Nrf2/HO-1 signaling）相关蛋白的表达水平。10 mg/kg剂量的α-香附酮可显著改善慢性肾脏病模型小鼠的肾功能，减轻肾小管间质纤维化，其干预效果与卡托普利相当。慢性肾脏病造模成功后，小鼠血清炎症介质水平、肾组织Ly6G表达量、超氧阴离子活性、ROS水平及8-OHdG水平均显著升高，而抗氧化酶SOD活性显著降低。α-香附酮或卡托普利给药均可改善上述所有异常病理生理变化。此外，α-香附酮可抑制慢性肾脏病模型小鼠肾组织中的NF-κB信号通路，并激活Akt/Nrf2/HO-1信号通路。α-香附酮可通过抑制炎症反应与氧化应激，经由NF-κB及Akt/Nrf2/HO-1信号通路改善慢性肾脏病模型小鼠的肾纤维化。