目的 探讨枸杞水提物（LBAE）对马兜铃酸Ⅰ（AAI）诱导HK-2细胞损伤的保护作用及其可能机制。方法 采用CCK-8法筛选AAI建模浓度及LBAE干预浓度。正式实验设置对照组、模型组、N-乙酰半胱氨酸（NAC，阳性药，5 mmol·L^-1）组和LBAE（800 μg·mL^-1）组，模型组和给药组给予100 μmol·L^-1 AAI造模，同时给予相应药物处理24 h，对照组加入等体积二甲基亚砜（DMSO）。采用CCK-8法检测细胞活力，检测培养上清乳酸脱氢酶（LDH）释放水平，倒置显微镜和透射电镜观察细胞形态及超微结构；试剂盒法检测超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、丙二醛（MDA）及肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-12水平；采用Annexin V-FITC/PI流式细胞术检测细胞凋亡； Western blotting检测凋亡相关蛋白及磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路相关蛋白表达。结果 100 μmol·L^-1 AAI可使HK-2细胞形成稳定损伤，800 μg·mL^-1 LBAE（以生药量计）和5 mmol·L^-1 NAC干预作用较明显，故分别作为后续建模及干预浓度。与对照组比较，模型组细胞活力显著降低（P＜0.01），LDH释放水平显著升高（P＜0.01），细胞形态及超微结构损伤明显；SOD、GSH水平显著降低，MDA及TNF-α、IL-6、IL-12水平显著升高（P＜0.01）；细胞总凋亡率显著升高（P＜0.01），B细胞淋巴瘤/白血病-2（Bcl-2）表达显著降低，Bcl-2相关X蛋白（Bax）及半胱氨酸蛋白水解酶（Caspase）相关蛋白表达显著升高（P＜0.01）； PI3K/Akt信号通路相关蛋白表达及其磷酸化比值显著降低（P＜0.01）。与模型组比较，LBAE干预后细胞活力显著升高（P＜0.01），LDH释放水平显著降低（P＜0.01），细胞形态及超微结构损伤减轻； SOD、GSH水平显著升高（P＜0.05、0.01），MDA及TNF-α、IL-6、IL-12水平显著降低（P＜0.05、0.01）；细胞总凋亡率显著降低（P＜0.01），Bcl-2表达显著升高（P＜0.05），Bax及Caspase相关蛋白表达显著降低（P＜0.01）； PI3K/Akt信号通路相关蛋白表达及其磷酸化比值显著升高（P＜0.05、0.01）。结论 LBAE可减轻AAI诱导的HK-2细胞损伤，其作用可能与改善氧化应激和炎症反应、抑制细胞凋亡及调节PI3K/Akt信号通路有关。

Objective To investigate the protective effect of Lycium barbarum aqueous extract (LBAE) against aristolochic acid I (AAI)-induced injury in HK-2 cells and its potential mechanisms. Methods CCK-8 assay was used to determine the optimal concentrations for AAI-induced model establishment and LBAE intervention. The formal experiment included a control group, a model group, an N-acetylcysteine (NAC, positive control drug at 5 mmol·L^-1) group, and an LBAE group (800 μg·mL^-1). The model and treatment groups were exposed to 100 μmol·L^-1 AAI for 24 h, with corresponding drugs administered simultaneously; the control group received an equal volume of dimethyl sulfoxide (DMSO). Cell viability was assessed by CCK-8 assay, LDH release levels in culture supernatants were measured, and cell morphology and ultrastructure were observed using inverted microscopy and transmission electron microscopy. Levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-12 were determined using commercial kits. Apoptosis was analyzed by Annexin V-FITC/PI flow cytometry, and expression of apoptosis-related proteins and PI3K/Akt signaling pathway proteins was detected by Western blotting. Results Exposure to 100 μmol·L^-1 AAI induced stable damage in HK-2 cells, while interventions with 800 μg·mL^-1 LBAE (based on crude drug weight) and 5 mmol·L^-1 NAC showed significant effects, thus these concentrations were selected for subsequent experiments. Compared with the control group, the model group exhibited significantly reduced cell viability (P < 0.01), markedly increased LDH release (P < 0.01), obvious morphological and ultrastructural damage, significantly decreased SOD and GSH levels, and significantly elevated MDA, TNF-α, IL-6, and IL-12 levels (P < 0.01). The total apoptosis rate was significantly higher (P < 0.01), Bcl-2 expression was significantly downregulated, whereas Bax and Caspase-related protein expressions were significantly upregulated (P < 0.01). Expression of PI3K/Akt signaling pathway proteins and their phosphorylation ratios were significantly reduced (P < 0.01). In comparison with the model group, LBAE treatment significantly improved cell viability (P < 0.01), reduced LDH release (P < 0.01), and alleviated cellular morphological and ultrastructural damage. SOD and GSH levels were significantly increased (P < 0.05, 0.01), while MDA, TNF-α, IL-6, and IL-12 levels were significantly decreased (P < 0.05, 0.01). The total apoptosis rate was significantly lower (P < 0.01), Bcl-2 expression was significantly enhanced (P < 0.05), and Bax and Caspase-related protein expressions were significantly suppressed (P < 0.01). Expression of PI3K/Akt signaling pathway proteins and their phosphorylation ratios were significantly increased (P < 0.05, 0.01). Conclusion LBAE can alleviate AAI-induced injury in HK-2 cells, possibly through ameliorating oxidative stress and inflammatory responses, inhibiting apoptosis, and modulating the PI3K/Akt signaling pathway.

R965

国家自然科学基金地区项目（82560859）；贵州省科技计划项目（黔科合基础-ZK［2024］一般366）；贵州中医药大学2025年校级研究生科研基金项目（YCXKYB2025004）