没食子酸表没食子儿茶素酯通过上调 miR-9 减轻尿酸诱导的大鼠肾间质成纤维细胞 NRK-49F 损伤
Epigallocatechin gallate attenuates uric acid-induced injury in rat renal interstitial fibroblasts NRK-49F by up-regulation of miR-9.
机构信息
Department of Nephrology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 Nov;22(21):7458-7469. doi: 10.26355/eurrev_201811_16287.
OBJECTIVE
Hyperuricemia is a common symptom in chronic kidney disease (CKD) and uric acid (UA) was observed to be elevated in CKD. Epigallocatechin gallate (EGCG) was reported to have multiple protective functions in inflammatory process. In this study, we investigated the effects of EGCG in UA-treated NRK-49F cells.
PATIENTS AND METHODS
NRK-49F cells induced by UA were treated with EGCG and/or transfected with microRNA-9 (miR-9) inhibitor or its negative control (NC). Cell viability and cell apoptosis were detected by Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. In addition, the expression of cell survival- and fibrosis-related factors was measured by qRT-PCR and Western blot, respectively. Concentrations of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA).
RESULTS
Increasing cell viability (p < 0.01) and decreasing cell apoptosis (p < 0.01), cell fibrosis (p < 0.01) and inflammatory cytokines (p < 0.05) were observed by administration of EGCG in UA-treated cells. In addition, miR-9 was significantly up-regulated by EGCG (p < 0.05 or p < 0.001). Furthermore, transfection with miR-9 inhibitor impaired the protective functions of EGCG in UA-treated NRK-49F cells (p < 0.05 or p < 0.01). EGCG significantly down-regulated expression of IκBα, p65, janus kinase (JAK) 2 and signal transducers and activator of transcription (STAT) 3 (all p < 0.05).
CONCLUSIONS
EGCG attenuates UA-induced injury in NRK-49F cells by up-regulation of miR-9 and might by inactivation of NF-κB and JAK-STAT signal pathways.
目的
高尿酸血症是慢性肾脏病（CKD）的常见症状，尿酸（UA）在 CKD 中升高。表没食子儿茶素没食子酸酯（EGCG）在炎症过程中具有多种保护功能。在这项研究中，我们研究了 EGCG 在 UA 处理的 NRK-49F 细胞中的作用。
患者和方法
用 UA 诱导的 NRK-49F 细胞用 EGCG 和/或 microRNA-9（miR-9）抑制剂或其阴性对照（NC）转染。通过细胞计数试剂盒-8（CCK-8）测定和流式细胞术分别检测细胞活力和细胞凋亡。此外，通过 qRT-PCR 和 Western blot 分别测量细胞存活和纤维化相关因子的表达。通过酶联免疫吸附试验（ELISA）检测炎症细胞因子的浓度。
结果
在 UA 处理的细胞中，给予 EGCG 可增加细胞活力（p < 0.01）和减少细胞凋亡（p < 0.01），细胞纤维化（p < 0.01）和炎症细胞因子（p < 0.05）。此外，EGCG 可显著上调 miR-9（p < 0.05 或 p < 0.001）。此外，miR-9 抑制剂的转染可损害 EGCG 在 UA 处理的 NRK-49F 细胞中的保护作用（p < 0.05 或 p < 0.01）。EGCG 显著下调 IκBα、p65、Janus 激酶（JAK）2 和信号转导和转录激活因子（STAT）3 的表达（均 p < 0.05）。
结论
EGCG 通过上调 miR-9 减轻 UA 诱导的 NRK-49F 细胞损伤，可能通过抑制 NF-κB 和 JAK-STAT 信号通路。
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