Xiang Haiyan, Wu Yan, Zhang Yun, Hong Yuanhao, Xu Yaling
Department of Nephrology, Wuhan Sixth Hospital, Affiliated Hospital of Jianghan University, No.168, Jiang 'an District, Wuhan, Hubei China.
Cytotechnology. 2024 Oct;76(5):559-569. doi: 10.1007/s10616-024-00638-x. Epub 2024 Jun 21.
Acute kidney injury (AKI) is a common clinical condition and is associated with unacceptable morbidity and mortality. Obtusifolin is an anthraquinone extracted from the seeds of with anti-inflammatory properties. This study focused on the role and mechanism of obtusifolin in AKI. The mouse podocyte cell line MPC5 was exposed to lipopolysaccharide (LPS) to establish a cell model of AKI. The viability of MPC5 cells treated with obtusifolin and/or LPS was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide assay. Cell apoptosis was analyzed by flow cytometry. The levels of podocyte injury- and apoptosis-related proteins as well as the nuclear factor-kappaB (NF-κB) signaling pathway was examined using western blotting analysis. The renal protective effects of obtusifolin were determined using an LPS-induced mouse model of AKI. Serum creatinine and blood urea nitrogen levels were measured. Hematoxylin-eosin staining of kidney sections was performed to evaluate renal histology. We found that MPC5 cells treated with LPS showed suppressed cell viability ( < 0.01) and increased cell apoptosis ( < 0.001). LPS reduced the protein expression of Bcl-2, nephrin, and synaptopodin as well as increased the protein levels of Bax and Cleaved Caspase-3 in podocytes in a concentration-dependent manner ( < 0.01). In addition, 10 μg/ml LPS-repressed cell viability was rescued by obtusifolin in a concentration-dependent manner ( < 0.01). Moreover, LPS-induced increase in MPC5 cell apoptosis was reversed by obtusifolin treatment ( < 0.01). Obtusifolin administration ameliorated LPS-induced kidney injury and reduced blood urea nitrogen and serum creatinine levels in mice ( < 0.001). Additionally, obtusifolin inhibited LPS-induced activation of NF-κB signaling in vitro and in vivo ( < 0.01). Overall, obtusifolin was effective in protecting renal function against LPS-induced AKI via inactivation of NF-κB signaling, which suggested that obtusifolin may act as a valuable agent for AKI therapy.
急性肾损伤（AKI）是一种常见的临床病症，与不可接受的发病率和死亡率相关。钝叶决明素是从钝叶决明种子中提取的一种具有抗炎特性的蒽醌。本研究聚焦于钝叶决明素在急性肾损伤中的作用及机制。将小鼠足细胞系MPC5暴露于脂多糖（LPS）以建立急性肾损伤细胞模型。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测经钝叶决明素和/或脂多糖处理的MPC5细胞的活力。通过流式细胞术分析细胞凋亡情况。使用蛋白质免疫印迹分析检测足细胞损伤和凋亡相关蛋白水平以及核因子-κB（NF-κB）信号通路。利用脂多糖诱导的急性肾损伤小鼠模型确定钝叶决明素的肾脏保护作用。检测血清肌酐和血尿素氮水平。对肾脏切片进行苏木精-伊红染色以评估肾脏组织学。我们发现，经脂多糖处理的MPC5细胞活力受到抑制（<0.01），细胞凋亡增加（<0.001）。脂多糖以浓度依赖的方式降低足细胞中Bcl-2、nephrin和突触素的蛋白表达，并增加Bax和裂解的半胱天冬酶-3的蛋白水平（<0.01）。此外，钝叶决明素以浓度依赖的方式挽救了10μg/ml脂多糖抑制的细胞活力（<0.01）。而且，钝叶决明素处理逆转了脂多糖诱导的MPC5细胞凋亡增加（<0.01）。给予钝叶决明素可改善脂多糖诱导的小鼠肾脏损伤，并降低小鼠血尿素氮和血清肌酐水平（<0.001）。此外，钝叶决明素在体外和体内均抑制脂多糖诱导的NF-κB信号激活（<0.01）。总体而言，钝叶决明素通过使NF-κB信号失活有效保护肾功能免受脂多糖诱导的急性肾损伤，这表明钝叶决明素可能是一种有价值的急性肾损伤治疗药物。
