背景:植酸苷(ACT)是源自地黄(地黄)叶子的主要成分。地黄液具有清热的功能,补气活血,中医滋阴补肾。最近的研究表明,地黄液可用于治疗肾炎,ACT是一种有前途的抗肾病药。
目的:阐明ACT在生物样品中的代谢产物,探讨ACT对慢性肾小球肾炎(CGN)大鼠肾脏的保护作用及其机制。
方法:在本研究中,通过四极杆飞行时间串联质谱法阐明了大鼠生物样品中ACT的生物转化。在肾病模型大鼠中通过尿液样本验证代谢物。通过高糖引起的肾小球足细胞损伤来评估ACT及其代谢物的作用。根据对体内成分的分析,通过网络药理学分析和分子对接研究CGN治疗的潜在治疗靶点。然后,在被动Heymann肾炎(PHN)模型中确定ACT的肾脏保护作用和机制。
结果:共检测并鉴定了49种ACT代谢物。同时,在肾病模型大鼠中检测到21种代谢物。ACT被迅速吸收并从肾脏转移,代谢产物通过尿液排出。整个过程持续约8小时。ACT对高糖损伤的肾小球足细胞有明显的保护作用,3,4-二羟基苯乙酸可能是ACT在体内的主要代谢产物。网络药理学和分子对接结果显示84个ACT-CGN靶点,其中MAPK1,HRAS,AKT1,EGFR,和其他人是高度相关的。在PHN大鼠模型中,ACT显著降低24小时尿蛋白和血清肌酐浓度,抑制白细胞CD18表达水平,降低血清肿瘤坏死因子α(TNF-α)水平,降低血清白细胞介素6(IL-6)水平。ACT显著降低血小板聚集率,抑制脾淋巴细胞对丝裂原伴刀豆球蛋白A的增殖活性。ACT抑制肾脏组织中转化生长因子-β和纤连蛋白的表达,并在1.8至1330μg/mL的剂量范围内剂量依赖性地抑制RAW264.7小鼠巨噬细胞中TNF-α和IL-6的产生。
结论:ACT对PHN大鼠有治疗作用,其机制可能与抑制细胞间或细胞间基质粘附有关,抑制炎症反应,调节免疫功能,改善组织血流动力学和血液流变学,和纤维化病变的缓解。
BACKGROUND: Acteoside (ACT) is the main ingredient derived from the leaves of Rehmannia glutinosa (Dihuangye). Dihuangye has the function of clearing heat, replenishing qi and activating blood, nourishing yin and tonifying kidney in traditional Chinese medicine. Recent studies have demonstrated that Dihuangye can be used to treat nephritis and ACT is a promising antinephritic agent.
OBJECTIVE: To clarify the metabolites of ACT in biological samples and investigate the renoprotective effect and mechanism of ACT in rats with chronic glomerulonephritis (CGN).
METHODS: In this study, the biotransformation of ACT in rat biological samples was clarified by quadrupole time-of-flight tandem mass spectrometry. The metabolites were validated by urine samples in nephropathy model rats. The effect of ACT and its metabolites was evaluated by glomerular podocyte injury due to high glucose. Based on an analysis of the ingredients in vivo, the potential therapeutic targets in the treatment of CGN were investigated by using network pharmacological analysis and molecular docking. Then, the renoprotective effect and mechanism of ACT were determined in rats in a passive Heymann nephritis (PHN) model.
RESULTS: A total of 49 metabolites of ACT were detected and identified. Meanwhile, 21 metabolites were detected in nephropathy model rats. ACT was absorbed rapidly and transferred from the kidney, and the metabolites were eliminated via urine. The whole process lasted approximately 8 h. ACT had a significant protective effect on glomerular podocytes damaged by high glucose and 3,4-dihydroxyphenylacetic acid might be the main metabolite of ACT underlying its functions in vivo. The network pharmacology and molecular docking results showed 84 ACT-CGN targets, among which MAPK1, HRAS, AKT1, EGFR, and others were a highly correlated. In the PHN rat model, ACT significantly reduced the 24-h urine protein and serum creatinine concentrations, suppressed the leukocyte CD18 expression levels, decreased the serum tumor necrosis factor α (TNF-α) levels and tended to reduce serum interleukin 6 (IL-6) levels. ACT significantly reduced the platelet aggregation rate and inhibited the proliferative activity of splenic lymphocytes in response to the mitogen concanavalin A. Meanwhile, ACT inhibited transforming growth factor-β and fibronectin expression in renal tissues and dose-dependently inhibited TNF-α and IL-6 production in RAW264.7 mouse macrophages at doses ranging from 1.8 to 1330 μg/mL.
CONCLUSIONS: ACT had therapeutic effects on PHN rats, and its mechanism might be related to the inhibition of intercellular or intercellular-matrix adhesion, suppression of inflammatory response, regulation of immune function, improvement of tissue hemodynamics and hemorheology, and relief of fibrotic lesions.