Department of Nephropathy, Dongguan Tungwah Hospital, No. 1 Dongcheng East Road, Dongguan, 523015, Guangdong Province, People's Republic of China.
Acta Diabetol. 2023 Apr;60(4):563-577. doi: 10.1007/s00592-022-02021-8. Epub 2023 Jan 20.
LncRNA SNHG16 and Toll-like receptor-4 (TLR4) participate in diabetes nephropathy. This study investigated whether SNHG16 regulates diabetic renal injury (DRI) via TLR4 and its related mechanism.
Diabetic mice and high glucose (HG)-induced HRMCs were used to examine the expressions of SNHG16 and TLR4. The SNHG16 expression, cytokines, reactive oxygen species, MDA, SOD, GSH, and fibrosis-related proteins were evaluated in HG-induced HRMCs transfected with sh-NC or sh-SHNG16. RNA immunoprecipitation and RNA pull-down determined the interaction between SNHG16 and EIF4A3 or TLR4 and EIF4A3. We used HG-treated HRMCs or diabetic mice to investigate the roles of TLR4 or SNHG16 in renal injuries.
Both SNHG16 and TLR4 were upregulated in diabetic conditions. HG increased serum Scr and BUN, led to significant fibrosis, increased inflammation- and renal fibrosis-related proteins in mice, and increased ROS, MDA, and decreased SOD and GSH in HRMCs. SNHG16 silencing diminished HG-upregulated SNHG16, decreased HG-increased cytokines secretion, ROS, MDA, and fibrosis but increased SOD and GSH. RIP and RNA pull-down confirmed that SNHG16 recruits EIF4A3 to stabilize TLR4 mRNA. TLR4 knockdown alleviated HG-induced renal injuries by suppressing RAS and NF-κB-mediated activation of NLRP3 inflammasomes. SNHG16 knockdown alleviated HG-induced renal injuries in HG-induced HRMCs or diabetic mice. Interestingly, TLR4 overexpression reversed the effects of SNHG16 knockdown. Mechanistically, SNHG16 knockdown alleviated HG-induced renal injuries by suppressing TLR4.
SNHG16 accelerated HG-induced renal injuries via recruiting EIF4A3 to enhance the stabilization of TLR4 mRNA. The SNGHG16/ELF4A3/TLR4 axis might be a novel target for treating DRI.
长链非编码 RNA SNHG16 和 Toll 样受体-4（TLR4）参与糖尿病肾病。本研究旨在探讨 SNHG16 是否通过 TLR4 及其相关机制调节糖尿病肾脏损伤（DRI）。
使用糖尿病小鼠和高糖（HG）诱导的人肾小球系膜细胞（HRMCs）来检测 SNHG16 和 TLR4 的表达。在转染 sh-NC 或 sh-SNHG16 的 HG 诱导的 HRMCs 中评估 SNHG16 表达、细胞因子、活性氧（ROS）、丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和纤维化相关蛋白。RNA 免疫沉淀和 RNA 下拉实验确定 SNHG16 与 EIF4A3 或 TLR4 与 EIF4A3 的相互作用。我们使用 HG 处理的 HRMCs 或糖尿病小鼠来研究 TLR4 或 SNHG16 在肾脏损伤中的作用。
在糖尿病条件下，SNHG16 和 TLR4 均上调。HG 增加了血清肌酐和血尿素氮，导致小鼠明显纤维化，增加了炎症和肾纤维化相关蛋白，同时增加了 HRMCs 中的 ROS、MDA，降低了 SOD 和 GSH。SNHG16 沉默减弱了 HG 上调的 SNHG16，减少了 HG 增加的细胞因子分泌、ROS、MDA，增加了 SOD 和 GSH。RNA 免疫沉淀和 RNA 下拉实验证实 SNHG16 招募 EIF4A3 来稳定 TLR4 mRNA。TLR4 敲低通过抑制 RAS 和 NF-κB 介导的 NLRP3 炎性小体激活来减轻 HG 诱导的肾损伤。SNHG16 敲低减轻了 HG 诱导的 HRMCs 或糖尿病小鼠的肾损伤。有趣的是，TLR4 过表达逆转了 SNHG16 敲低的作用。机制上，SNHG16 敲低通过抑制 TLR4 减轻了 HG 诱导的肾损伤。
SNHG16 通过招募 EIF4A3 增强 TLR4 mRNA 的稳定性，加速 HG 诱导的肾损伤。SNHG16/ELF4A3/TLR4 轴可能是治疗 DRI 的一个新靶点。
