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Abstract：
Obcordata A (OA) is a polyoxypregnane glycoside derived from the Dai medicine Aspidopterys obcordata vines. This study aims to investigate the efficacy of OA on renal tubular epithelial cells exposed to calcium oxalate crystals. We incubated renal tubular cells with 28 μg·cm2 calcium oxalate crystals for 24 h with and without OA, GKT137831, phorbol-12-myristate-13-acetate (PMA), and tocopherol. The MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, microscopic examination, flow cytometry, and immunofluorescence staining revealed that calcium oxalate crystals decreased cell viability and elevated reactive oxygen species (ROS) levels. OA, GKT137831, and tocopherol protected cells and decreased ROS levels. However, OA did not exhibit direct DPPH scavenging ability. In addition, immunoblotting illustrated that OA inhibited the NOX4 (nicotinamide adenine dinucleotide phosphate oxidases 4) expression and downregulated the protein expression in the NOX4/ROS/p38 MAPK (p38 mitogen-activated protein kinase) pathway. The findings suggest that the cytoprotective and antioxidant effects of OA can be blocked by the NOX4 agonist PMA. In conclusion, OA could be used as a NOX4 inhibitor to prevent kidney stones.
摘要：
ObcordataA（OA）是一种聚氧孕烷糖苷，来自the药Aspidopterysobcordatavines。本研究旨在探讨OA对暴露于草酸钙晶体的肾小管上皮细胞的功效。我们在有或没有OA的情况下，将肾小管细胞与28μg·cm2草酸钙晶体一起孵育24小时，GKT137831，佛波醇-12-肉豆蔻酸-13-乙酸酯（PMA），和生育酚.MTT[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物]测定,显微镜检查,流式细胞术,免疫荧光染色显示草酸钙晶体降低了细胞活力并升高了活性氧（ROS）水平。OA,GKT137831和生育酚保护细胞并降低ROS水平。然而,OA没有表现出直接的DPPH清除能力。此外,免疫印迹表明，OA抑制NOX4（烟酰胺腺嘌呤二核苷酸磷酸氧化酶4）的表达，并下调NOX4/ROS/p38MAPK（p38丝裂原活化蛋白激酶）途径中的蛋白质表达。研究结果表明，NOX4激动剂PMA可以阻断OA的细胞保护和抗氧化作用。总之,OA可用作NOX4抑制剂以预防肾结石。
