■足细胞损伤是糖尿病肾病(DKD)和肥胖相关性肾小球病(ORG)的常见病理机制。我们先前的研究证实,桦褐孔菌(IO)改善了DKD大鼠的足细胞损伤。目前的研究探索了药理作用,IO对ORG小鼠的相关机制及可能的活性成分。
■首先,通过超高效液相色谱-串联质谱分析(UPLC-Q-TOF-MS)与网络药理学相结合,构建人蛋白-蛋白相互作用机制,这导致发现了IO对抗ORG的关键机制。然后,通过高脂饮食和生化测定建立ORG小鼠,组织病理学,采用Westernblot方法探讨IO对肥胖及足细胞损伤的影响。最后,基于网络药理学的发现通过免疫组织化学得到证实.通过UPLC-Q-TOF-MS分析小鼠血浆中吸收的IO的组成,并使用分子对接来预测可能的活性化合物。
■网络药理学结果提示IO通过调节TNF信号减轻ORG的炎症反应。20周的体内实验证实,IO改善肾小球肥大,电镜下足细胞损伤,肾nephrin,突触素,肿瘤坏死因子-α和白细胞介素-6的表达采用免疫印迹和免疫组织化学染色。ORG的其他指标,如体重,肾脏重量,血清总胆固醇,IO干预也改善了肝脏甘油三酯。组分分析显示,三萜,包括异萜烯F和三甲烯醇酸,可能是药效学基础。
■基于UPLC-Q-TOF-MS分析的研究,网络药理学和体内实验表明,IO通过调节TNF信号改善ORG小鼠足细胞损伤。三萜类化合物被预测为作用成分。
UNASSIGNED: Podocyte injury is a common pathologic mechanism in diabetic kidney disease (DKD) and obesity-related glomerulopathy (ORG). Our previous study confirmed that Inonotus obliquus (IO) improved podocyte injury on DKD rats. The current study explored the pharmacological effects, related mechanisms and possible active components of IO on ORG mice.
UNASSIGNED: Firstly, by combining ultra-high performance liquid chromatography tandem mass spectrometry analysis (UPLC-Q-TOF-MS) with network pharmacology to construct the human protein-protein interaction mechanism and enrich the pathway, which led to discover the crucial mechanism of IO against ORG. Then, ORG mice were established by high-fat diet and biochemical assays, histopathology, and Western blot were used to explore the effects of IO on obesity and podocyte injury. Finally, network pharmacology-based findings were confirmed by immunohistochemistry. The compositions of IO absorbed in mice plasma were analyzed by UPLC-Q-TOF-MS and molecular docking was used to predict the possible active compounds.
UNASSIGNED: The network pharmacology result suggested that IO alleviated the inflammatory response of ORG by modulating TNF signal. The 20-week in vivo experiment confirmed that IO improved glomerular hypertrophy, podocyte injury under electron microscopy, renal nephrin, synaptopodin, TNF-α and IL-6 expressions with Western blotting and immunohistochemical staining. Other indicators of ORG such as body weight, kidney weight, serum total cholesterol, liver triglyceride also improved by IO intervention. The components analysis showed that
triterpenoids, including inoterpene F and trametenolic acid, might be the pharmacodynamic basis.
UNASSIGNED: The research based on UPLC-Q-TOF-MS analysis, network pharmacology and in vivo experiment suggested that the amelioration of IO on podocyte injury in ORG mice via its modulation on TNF signal.
Triterpenoids were predicated as acting components.