Department of Anatomy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
School of Medicine, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Biomed Res Int. 2019 Sep 22;2019:5423703. doi: 10.1155/2019/5423703. eCollection 2019.
Kidney ischemic/reperfusion (I/R) injury is the main cause of acute kidney injury (AKI) involving renal function deterioration, renal architecture damage, and inflammation. This condition may lead to kidney fibrosis with epithelial to mesenchymal transition (EMT) and myofibroblast formation. Inhibition of chronic effects of kidney I/R injury may provide effective strategies for treating AKI and chronic kidney diseases (CKDs). Chlorogenic acid (CGA) is recognized as a powerful antioxidant, with anti-inflammatory and antifibrotic properties in many conditions. However, the effect of CGA on kidney I/R injury has not been elucidated yet. Kidney I/R injury was performed on male Swiss background mice (I/R group, = 5, 3-4 months, 30-40 g) which underwent bilateral renal pedicles clamping for 30 minutes and then were euthanized on day three after operation. Three groups of I/R were treated with 3 different doses of CGA intraperitoneally for 2 days: 3.5 (I/R + CGA1 group), 7 (I/R + CGA2 group), and 14 (I/R + CGA3 group) mg/kg of body weight. Tubular injury was quantified based on Periodic Acid-Schiff staining, while reverse transcriptase PCR (RT-PCR) was performed to quantify mRNA expression of TGF-1, vimentin, SOD-1, TLR-4, TNF-α, NF-κB and MCP-1. Immunohistochemical staining was done to quantify proliferating cell nuclear antigen (PCNA), myofibroblast (-SMA), SOD-1 and macrophage (CD68) number. Kidney I/R demonstrated tubular injury and increased inflammatory mediator expression, macrophage number, and myofibroblast expansion. Meanwhile, histological analysis showed lower tubular injury with higher epithelial cell proliferation in CGA-treated groups compared to the I/R group. RT-PCR also revealed significantly lower TGF-1 and vimentin mRNA expressions with higher SOD-1 mRNA expression. CGA-treated groups also demonstrated a significantly lower macrophage and myofibroblast number compared to the I/R group. These findings associated with lower mRNA expression of TLR-4, TNF-α, NF-κB, and MCP-1 as inflammatory mediators in CGA groups. I/R + CGA3 represented the highest amelioration effect among other CGA-treated groups. CGA treatment attenuates kidney I/R injury through reducing inflammation, decreasing myofibroblast expansion, and inducing epithelial cells proliferation.
肾缺血/再灌注（I/R）损伤是导致急性肾损伤（AKI）的主要原因，其涉及肾功能恶化、肾结构损伤和炎症。这种情况可能导致肾纤维化，伴有上皮到间充质转化（EMT）和肌成纤维细胞形成。抑制肾 I/R 损伤的慢性影响可能为治疗 AKI 和慢性肾脏病（CKD）提供有效的策略。绿原酸（CGA）被认为是一种强大的抗氧化剂，在许多情况下具有抗炎和抗纤维化的特性。然而，CGA 对肾 I/R 损伤的影响尚未阐明。在雄性瑞士背景小鼠（I/R 组，n=5，3-4 个月，30-40g）上进行肾 I/R 损伤，对双侧肾蒂进行 30 分钟夹闭，然后在手术后第 3 天处死。三组 I/R 分别用 3 种不同剂量的 CGA 进行腹腔内治疗 2 天：3.5（I/R+CGA1 组）、7（I/R+CGA2 组）和 14（I/R+CGA3 组）mg/kg 体重。通过过碘酸希夫染色定量肾小管损伤，同时通过逆转录 PCR（RT-PCR）定量 TGF-1、波形蛋白、SOD-1、TLR-4、TNF-α、NF-κB 和 MCP-1 的 mRNA 表达。进行免疫组织化学染色以定量增殖细胞核抗原（PCNA）、肌成纤维细胞（-SMA）、SOD-1 和巨噬细胞（CD68）的数量。肾 I/R 表现出肾小管损伤和炎症介质表达增加、巨噬细胞数量和肌成纤维细胞扩张。同时，组织学分析显示 CGA 治疗组的肾小管损伤较低，上皮细胞增殖较高。RT-PCR 还显示 TGF-1 和波形蛋白 mRNA 表达显著降低，SOD-1 mRNA 表达升高。CGA 治疗组与 I/R 组相比，巨噬细胞和肌成纤维细胞数量也显著降低。这些发现与 TLR-4、TNF-α、NF-κB 和 MCP-1 等炎症介质的 mRNA 表达降低有关。CGA 治疗组中，CGA+3 组的改善效果最高。CGA 治疗通过减少炎症、减少肌成纤维细胞扩张和诱导上皮细胞增殖来减轻肾 I/R 损伤。
