Chen Li, Xu Tangdi, Wang Jiahao, Wang Zixuan, Pan Ying, Kong Lingdong
State Key Laboratory of Pharmaceutical Biotechnology, Institute of Chinese Medicine, Nanjing Drum Tower Hospital, School of Life Sciences, Nanjing University, Nanjing, PR China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 2):118878. doi: 10.1016/j.jep.2024.118878. Epub 2024 Oct 1.
Siwu tablet (SWT), derived from a traditional Chinese medicinal formula named Siwu decoction, is widely used for blood deficiency syndrome. Siwu decoction and its derived formulas have been proven to improve renal anemia and prevent senescence. Whether SWT prevents glomerular podocyte senescence and the underlying molecular mechanism remains unknow.
To elucidate the protective effect and possible mechanism of SWT on glomerular podocyte senescence.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed to characterize components of SWT. Male Sprague-Dawley rats were given 10% fructose drinking water for 16 weeks. SWT (810 and 1620 mg/kg) was administered orally for the last 8 weeks. The assays of senescence-associated beta-galactosidase (SA-β-gal) staining, immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot as well as enzyme linked immunosorbent assay were performed to evaluate rat glomerular podocyte senescence. The mRNA and protein levels of nucleoporin 155 (Nup155) and inositol requiring mutant 80 (INO80) in rat glomeruli were detected by qRT-PCR, Western blot and immunofluorescence. Foot processes and nuclear pore complexes (NPCs) of rat glomerular podocytes were visualized by transmission electron microscopy.
One hundred and fifty-nine components were preliminarily identified in SWT. The results of animal experiments showed that SWT decreased the activity of SA-β-gal, protein levels of p16, p21, p53 and phosphorylated histone H2AX (γ-H2AX), and mRNA levels of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) in glomeruli of high fructose-fed rats. As expected, SWT increased renal cortex erythropoietin mRNA expression and serum erythropoietin concentration in this animal model. SWT reduced urine albumin-to-creatinine ratio and serum levels of uric acid, creatinine and blood urea nitrogen, and recovered glomerular structure injury in high fructose-fed rats. It up-regulated mRNA and protein levels of Nup155 and the number of podocyte NPCs, and subsequently reinforced mRNA nuclear export and protein expression of INO80 in rat glomeruli under high fructose stimulation.
SWT ameliorates glomerular podocyte senescence in high fructose-fed rats possibly by increasing Nup155 to promote INO80 mRNA nuclear export.
四物片（SWT）源自传统中药方剂四物汤，广泛用于治疗血虚证。四物汤及其衍生方剂已被证明可改善肾性贫血并预防衰老。SWT是否能预防肾小球足细胞衰老及其潜在分子机制尚不清楚。
阐明SWT对肾小球足细胞衰老的保护作用及可能机制。
采用液相色谱-串联质谱（LC-MS/MS）对SWT的成分进行表征。雄性Sprague-Dawley大鼠饮用10%果糖水16周。在最后8周口服给予SWT（810和1620mg/kg）。进行衰老相关β-半乳糖苷酶（SA-β-gal）染色、免疫组织化学、定量实时聚合酶链反应（qRT-PCR）、蛋白质印迹以及酶联免疫吸附测定等实验，以评估大鼠肾小球足细胞衰老情况。通过qRT-PCR、蛋白质印迹和免疫荧光检测大鼠肾小球中核孔蛋白155（Nup155）和肌醇需求突变体80（INO80）的mRNA和蛋白质水平。通过透射电子显微镜观察大鼠肾小球足细胞的足突和核孔复合体（NPC）。
在SWT中初步鉴定出159种成分。动物实验结果表明，SWT降低了高果糖喂养大鼠肾小球中SA-β-gal的活性、p16、p21、p53和磷酸化组蛋白H2AX（γ-H2AX）的蛋白质水平，以及白细胞介素-1β（IL-1β）、IL-6和肿瘤坏死因子-α（TNF-α）的mRNA水平。不出所料，在该动物模型中，SWT增加了肾皮质促红细胞生成素mRNA表达和血清促红细胞生成素浓度。SWT降低了高果糖喂养大鼠的尿白蛋白与肌酐比值以及血清尿酸、肌酐和血尿素氮水平，并恢复了肾小球结构损伤。在高果糖刺激下，SWT上调了大鼠肾小球中Nup155的mRNA和蛋白质水平以及足细胞NPC数量，随后增强了INO80的mRNA核输出和蛋白质表达。
SWT可能通过增加Nup155以促进INO80 mRNA核输出，改善高果糖喂养大鼠的肾小球足细胞衰老。
