摘要
背景:近年来,由嘌呤代谢紊乱引起的高尿酸血症的发病率不断增加,可诱导炎症反应并导致肾损伤。目的:探讨溶质载体家族2成员12(solute carrier family 2 member 12,SLC2A12)在高尿酸相关肾损伤中的作用和机制。方法:将肾小管细胞(HK2细胞)分为5组:HK2组、HK2+尿酸组、HK2+尿酸+NC组、HK2+尿酸+siSLC2A12组、HK2+尿酸+siSLC2A12+MK-2206组。用尿酸处理HK2细胞后转染siRNA SLC2A12,再通过MK-2206处理抑制AKT表达。采用CCK-8检测细胞增殖能力,TUNEL检测细胞凋亡情况,qRT-PCR和Western blot检测纤维化因子以及AKT/FOXO3a途径相关因子的表达,酶联免疫吸附实验检测细胞上清液中炎性细胞因子水平。结果与结论:①与HK2组比较,HK2+尿酸组细胞增殖能力降低,细胞凋亡增多,HK2+尿酸+siSLC2A12组细胞增殖能力进一步降低,凋亡细胞进一步增多;与HK2+尿酸+siSLC2A12组比较,HK2+尿酸+siSLC2A12+MK-2206组细胞增殖能力升高,凋亡细胞减少;②与HK2组比较,HK2+尿酸组细胞中结缔组织生长因子、α-平滑肌肌动蛋白、转化生长因子β表达升高,HK2+尿酸+siSLC2A12组3种因子表达进一步升高;与HK2+尿酸+siSLC2A12组比较,HK2+尿酸+siSLC2A12+MK-2206组3种因子表达有所下降;③与HK2组比较,HK2+尿酸组p-AKT、FOXO3a、p-FOXO3a表达升高,HK2+尿酸+siSLC2A12组p-AKT表达进一步升高,而FOXO3a、p-FOXO3a表达下降;与HK2+尿酸+siSLC2A12组比较,HK2+尿酸+siSLC2A12+MK-2206组p-AKT表达下降,FOXO3a、p-FOXO3a表达升高;④与HK2组比较,HK2+尿酸组白细胞介素6、白细胞介素1β、肿瘤坏死因子α分泌水平升高,HK2+尿酸+siSLC2A12组白细胞介素6、白细胞介素1β、肿瘤坏死因子α水平进一步升高;与HK2+尿酸+siSLC2A12组比较,HK2+尿酸+siSLC2A12+MK-2206组白细胞介素6、白细胞介素1β、肿瘤坏死因子α水平下降;⑤结果表明,SLC2A12可能通过激活FOXO3a途径抵抗尿酸诱导的肾小管纤维化及炎症反应,从而保护高尿酸血症诱�
BACKGROUND:In recent years,the incidence of hyperuricemia caused by purine metabolism disorders has been increasing,which can induce inflammatory responses and lead to renal injury.OBJECTIVE:To explore the role and mechanism of solute carrier family 2 member 12(SLC2A12)in hyperuricemia-related renal injury.METHODS:Renal tubular cells(HK2 cells)were divided into five groups:HK2 group,HK2+uric acid group,HK2+uric acid+NC group,HK2+uric acid+siSLC2A12 group,and HK2+uric acid+siSLC2A12+MK-2206 group.HK2 cells were treated with uric acid and transfected with siRNA SLC2A12,followed by MK-2206 treatment to inhibit AKT expression.Cell proliferation was detected by CCK-8 assay.Apoptosis was detected by TUNEL assay.qRT-PCR and western blot assay were used to detect fibrogenic factors as well as activation of the AKT/FOXO3a pathway.The concentrations of inflammatory cytokines were measured by enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:(1)Uric acid treatment inhibited cell proliferation and promoted cell apoptosis in the HK2+uric acid group compared with the HK2 group.The proliferative ability of cells in the HK2+uric acid+siSLC2A12 group was further decreased and apoptotic cells were further increased compared with the HK2 group.Compared with the HK2+uric acid+siSLC2A12 group,the HK2+uric acid+siSLC2A12+MK-2206 group showed an increase in cell proliferation and a decrease in apoptotic cells.(2)Compared with the HK2 group,the connective tissue growth factor(CTGF),α-smooth muscle actin(α-SMA)and transforming growth factor beta(TGF-β)expressions increased in the HK2+uric acid group;CTGF,α-SMA and TGF-βexpression further increased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,the CTGF,α-SMA and TGF-βexpressions decreased.(3)Compared with the HK2 group,the expression of p-AKT,FOXO3a,and p-FOXO3a elevated in the HK2+uric acid group;the expression of p-AKT further increased,while the expression of FOXO3a and p-FOXO3a decreased in the HK2+uric acid+siSLC2A12 group.Compared
作者
贺怡
李晓林
何金科
蒋香菊
梁美婷
陈邬锦
崔月娜
孙玉萍
He Yi;Li Xiaolin;He Jinke;Jiang Xiangju;Liang Meiting;Chen Wujin;Cui Yuena;Sun Yuping(Xinjiang Second Medical College,Karamay 834000,Xinjiang Uygur Autonomous Region,China;School of Basic Medical Sciences,Xinjiang Medical University,Urumqi 830000,Xinjiang Uygur Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第13期2076-2081,共6页
Chinese Journal of Tissue Engineering Research
基金
新疆维吾尔自治区高校科研计划(XJEDU2021Y054),项目负责人:贺怡~。
