■糖尿病中的高血糖通过非酶反应增加了糖基化终产物(AGEs)的产生。AGEs与其受体(RAGE)之间的相互作用导致氧化和炎症应激,在糖尿病肾病的发展中起着关键作用。香umi(SC)L.(DC。)顺势疗法制剂,即。(200C,30C,和母亲酊剂[OT])用于治疗糖尿病。
目的:本研究旨在阐明SC制剂的调节作用(200C,30C,和MT)对核因子红细胞2相关因子2(Nrf2)-核因子κB(NF-κB)途径和线粒体功能障碍减轻糖尿病肾病。
方法:链脲佐菌素诱导的糖尿病大鼠用SC制剂治疗(200C,30C,MT;在蒸馏水中1:20稀释;600μL/kg体重)和二甲双胍(45mg/kg体重),每天两次,持续40天。DN通过生化参数和组织学检查进行评估。分析肾组织裂解物的糖化标志物。Nrf2,NF-κB的蛋白质和基因水平,通过蛋白质印迹和RT-qPCR确定线粒体功能失调信号。对肾脏进行免疫组织化学分析。体外,人血清白蛋白(HSA-10mg/ml)用甲基乙二醛(MGO-55mM)在SC制剂存在下进行糖化(200C,30C,MT)八天。将糖化样品(400μg/mL)与肾细胞(HEK-293)一起孵育24小时。进一步生产活性氧,Nrf2核易位,蛋白质印迹分析Nrf2和凋亡标志物的蛋白质或基因表达,RT-qPCR,和流式细胞术。进行没食子酸和鞣花酸与HSA-MGO复合物的分子对接。
结果:使用链脲佐菌素诱导的糖尿病大鼠用SC制剂治疗的体内实验显示出改善的生化参数,保留肾功能,并以剂量依赖性方式减少糖化加合物的形成。此外,SC制剂下调炎症介质,如RAGE,NF-κB,血管内皮生长因子(VEGF),和肿瘤坏死因子α(TNF-α),同时上调Nrf2依赖性抗氧化剂和解毒途径。他们下调B细胞淋巴瘤2(Bcl-2)相关X蛋白(BAX),C/EBP同源蛋白(CHOP),动力蛋白相关蛋白1(DRP1),并上调BCL2基因表达。值得注意的是,SC制剂促进了Nrf2的核易位,导致抗氧化酶的上调和氧化应激标志物的下调。分子对接研究揭示了没食子酸(-5.26kcal/mol)和鞣花酸(-4.71kcal/mol)与HSA-MGO复合物之间的有利相互作用。
结论:SC制剂通过Nrf2依赖性机制减轻肾细胞凋亡和线粒体功能障碍。
UNASSIGNED: Hyperglycemia in diabetes increases the generation of advanced
glycation end products (AGEs) through non-enzymatic reactions. The interaction between AGEs and their receptors (RAGE) leads to oxidative and inflammatory stress, which plays a pivotal role in developing diabetic nephropathy. Syzygium cumini (SC) L. (DC.) homeopathic preparations viz. (200C, 30C, and mother tincture [MT]) are used to treat diabetes.
OBJECTIVE: This study aimed to elucidate the regulatory effects of SC preparations (200C, 30C, and MT) on the nuclear factor erythroid 2-related factor 2 (Nrf2) - nuclear factor-κB (NF-κB) pathways and mitochondrial dysfunction in mitigating Diabetic nephropathy.
METHODS: Streptozotocin-induced diabetic rats were treated with SC preparations (200C, 30C, MT; 1:20 dilution in distilled water; 600 μL/kg body weight) and metformin (45 mg/kg body weight) twice daily for 40 days. DN was evaluated through biochemical parameters and histological examination. Renal tissue lysates were analyzed for
glycation markers. Protein and gene levels of Nrf2, NF-κB, and mitochondrial dysfunctional signaling were determined via western blotting and RT-qPCR. An immunohistochemical analysis of the kidneys was performed. In vitro, human serum albumin (HSA - 10 mg/ml) was glycated with methylglyoxal (MGO - 55 mM) in the presence of SC preparations (200C, 30C, MT) for eight days. Glycated samples (400 μg/mL) were incubated with renal cells (HEK-293) for 24 hours. Further reactive oxygen species production, Nrf2 nuclear translocation, and protein or gene expression of Nrf2 and apoptosis markers were analyzed by western blotting, RT-qPCR, and flow cytometry. Molecular docking of gallic and ellagic acid with the HSA-MGO complex was performed.
RESULTS: In vivo experiments using streptozotocin-induced diabetic rats treated with SC preparations exhibited improved biochemical parameters, preserved kidney function, and reduced
glycation adduct formation in a dose-dependent manner. Furthermore, SC preparations downregulated inflammatory mediators such as RAGE, NF-κB, vascular endothelial growth factor (VEGF), and Tumor necrosis factor α (TNF-α) while upregulating the Nrf2-dependent antioxidant and detoxification pathways. They downregulated B-cell lymphoma 2 (Bcl-2) associated X-protein (BAX), C/EBP homologous protein (CHOP), Dynamin-related protein 1 (DRP1), and upregulated BCL 2 gene expression. Notably, SC preparations facilitated nuclear translocation of Nrf2, leading to the upregulation of antioxidant enzymes and the downregulation of oxidative stress markers. Molecular docking studies revealed favorable interactions between gallic (-5.26 kcal/mol) and ellagic acid (-4.71 kcal/mol) with the HSA-MGO complex.
CONCLUSIONS: SC preparations mitigate renal cell apoptosis and mitochondrial dysfunction through Nrf2-dependent mechanisms.