穿心莲内酯对肾小管间质性损伤和纤维化的影响。其作用机制的证据。
Effects of andrographolide on renal tubulointersticial injury and fibrosis. Evidence of its mechanism of action.
机构信息
Guangdong Pharmaceutical University, Guangzhou 510006, China; Guangdong Metabolic Diseases Research Center of Integrated Chinese and Western Medicine, Guangzhou 510006, China; Key Laboratory of Glucolipid Metabolic Disorder, Ministry of Education, Guangzhou 510006, China; Institute of Chinese Medicine, Guangdong TCM Key Laboratory for Metabolic Diseases, Guangzhou 510006, China.
Department of Nephrology, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou 510632, China.
出版信息
Phytomedicine. 2021 Oct;91:153650. doi: 10.1016/j.phymed.2021.153650. Epub 2021 Jul 9.
BACKGROUND
Diabetic nephropathy (DN) is associated with renal interstitial injury and fibrosis. Our previous study showed that andrographolide protected against the progression of DN and high glucose (HG)-induced mesangial dysfunction. However, the protective effects of andrographolide on renal tubular epithelial cells have not been fully elucidated.
PURPOSE
To determine the protective effects of andrographolide on renal tubular damage and explore the underlying mechanism.
STUDY DESIGN
Human tubular epithelial cells (HK-2 cells) were treated with andrographolide (5 and 10 μM) under HG conditions. Diabetic mice were treated with andrographolide (i.p. 2 and 4 mg/kg, twice per week).
METHODS
Western blotting, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence and flow cytometry were used to analyze the effects of andrographolide on renal tubular injury and fibrotic mechanisms in HK-2 cells. The protective effects of andrographolide against renal tubulointerstitial injury and fibrosis were investigated in diabetic mice fed a high-fat diet (HFD). Renal interstitial tissue was collected at sacrifice for immunohistochemistry, immunofluorescence analysis, RT-PCR and Western blotting to analyze the effects of andrographolide on renal tubular injury and fibrosis.
RESULTS
In vitro assay results indicated that andrographolide (5 and 10 μM) effectively inhibited HG-induced apoptosis, epithelial-mesenchymal transition (EMT) and collagen deposition in HK-2 cells. Mechanistically, HG stimulated mitochondrial reactive oxygen species (mtROS)-mediated NOD-like receptor family and pyrin domain-containing protein 3 (NLRP3) inflammasome activation and EMT in tubular epithelial cells, and andrographolide (5 and 10 μM) inhibited these effects by ameliorating mitochondrial dysfunction. In vivo, treatment with andrographolide (2 and 4 mg/kg) inhibited renal tubular cell apoptosis, EMT and tubulointerstitial fibrosis, mitochondrial dysfunction and NLRP3 inflammasome activation in diabetic mice.
CONCLUSION
Andrographolide (5 and 10 μM) prevents HG-induced renal tubular cell damage, and andrographolide (2 and 4 mg/kg) protects against the progression of diabetic tubular injury and fibrosis in mice by suppressing mitochondrial dysfunction and NLRP3 inflammasome activation.
背景
糖尿病肾病（DN）与肾间质损伤和纤维化有关。我们之前的研究表明，穿心莲内酯可防止 DN 和高糖（HG）诱导的系膜功能障碍的进展。然而，穿心莲内酯对肾小管上皮细胞的保护作用尚未完全阐明。
目的
确定穿心莲内酯对肾小管损伤的保护作用，并探讨其潜在机制。
研究设计
在 HG 条件下，用穿心莲内酯（5 和 10 μM）处理人肾小管上皮细胞（HK-2 细胞）。糖尿病小鼠用穿心莲内酯（腹腔注射 2 和 4 mg/kg，每周两次）治疗。
方法
用 Western blot、逆转录聚合酶链反应（RT-PCR）、免疫荧光和流式细胞术分析穿心莲内酯对 HK-2 细胞肾小管损伤和纤维化机制的影响。在高脂饮食（HFD）喂养的糖尿病小鼠中研究穿心莲内酯对肾小管间质性损伤和纤维化的保护作用。处死时收集肾间质组织进行免疫组织化学、免疫荧光分析、RT-PCR 和 Western blot，以分析穿心莲内酯对肾小管损伤和纤维化的影响。
结果
体外实验结果表明，穿心莲内酯（5 和 10 μM）可有效抑制 HG 诱导的 HK-2 细胞凋亡、上皮-间充质转化（EMT）和胶原沉积。机制上，HG 刺激线粒体活性氧（mtROS）介导的 NOD 样受体家族和吡啶结构域蛋白 3（NLRP3）炎性小体激活和肾小管上皮细胞 EMT，穿心莲内酯（5 和 10 μM）通过改善线粒体功能障碍抑制这些作用。在体内，穿心莲内酯（2 和 4 mg/kg）治疗可抑制糖尿病小鼠肾小管细胞凋亡、EMT 和肾小管间质性纤维化、线粒体功能障碍和 NLRP3 炎性小体激活。
结论
穿心莲内酯（5 和 10 μM）可防止 HG 诱导的肾小管细胞损伤，穿心莲内酯（2 和 4 mg/kg）通过抑制线粒体功能障碍和 NLRP3 炎性小体激活来防止糖尿病肾小管损伤和纤维化的进展。
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