Lin Jian, Feng Jian-Bo, Su Jing-Lin, Lin Nan, Cai Yin-Sheng, Lv Fei
Department of Clinical Laboratory, Dongguan Tungwah Hospital, Dongguan, 523129, Guangdong, China.
Department of Clinical Laboratory, Dongguan SongShan Lake Tungwah Hospital, No. 1, Kefa Seventh Road, Songshan Lake Science and Technology Park, Dongguan, 523808, Guangdong, China.
Clin Exp Nephrol. 2025 Jul 14. doi: 10.1007/s10157-025-02718-4.
Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. We aimed to explore the role of RNA binding motif protein 15 (RBM15) in high glucose (HG)-induced pyroptosis of renal tubular epithelial cell, thus providing theoretical knowledge and new targets for DN treatment.
HG-induced HK-2 cells were used to establish DN cell models. RBM15 expression was inhibited in HK-2 and detected. Cell viability was detected by cell counting kit-8. The levels of NLR family pyrin domain containing 3 (NLRP3), NLR family CARD domain containing 4 (NLRC4), gasdermin D (GSDMD)-N, and cleaved Caspase-1 were detected by Western blot assay. The levels of IL-1β and IL-18 were detected by enzyme linked immunosorbent assay. The enrichment of insulin-like growth factor 2 mRNA binding protein (IGF2BP2) and N6-methyladenosine (m6A) on NLRP3 and NLRC4 were analyzed by methylated RNA immunoprecipitation (MeRIP) and RIP. The stability of NLRP3 and NLRC4 mRNA was analyzed. The mechanism was verified by interfering NLRP3 and NLRC4 expression.
RBM15 was highly expressed in HG-induced HK-2 cells. Inhibition of RBM15 reversed cell viability, inhibited inflammation, and alleviated pyroptosis. RBM15 promoted the expression of inflammasomes NLRP3 and NLRC4 through IGF2BP2-dependent m6A modification.
RBM15 promoted the expression of inflammasomes NLRP3 and NLRC4 through IGF2BP2-dependent m6A modification, thereby promoting pyroptosis.
糖尿病肾病（DN）是终末期肾病的主要病因。我们旨在探讨RNA结合基序蛋白15（RBM15）在高糖（HG）诱导的肾小管上皮细胞焦亡中的作用，从而为DN治疗提供理论依据和新靶点。
用HG诱导的HK-2细胞建立DN细胞模型。抑制HK-2细胞中RBM15的表达并进行检测。采用细胞计数试剂盒-8检测细胞活力。通过蛋白质免疫印迹法检测含NLR家族吡啉结构域3（NLRP3）、含NLR家族CARD结构域4（NLRC4）、gasdermin D（GSDMD）-N和裂解的半胱天冬酶-1的水平。采用酶联免疫吸附测定法检测白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）的水平。通过甲基化RNA免疫沉淀（MeRIP）和RNA免疫沉淀（RIP）分析胰岛素样生长因子2 mRNA结合蛋白（IGF2BP2）和N6-甲基腺苷（m6A）在NLRP3和NLRC4上的富集情况。分析NLRP3和NLRC4 mRNA的稳定性。通过干扰NLRP3和NLRC4的表达验证其机制。
RBM15在HG诱导的HK-2细胞中高表达。抑制RBM15可逆转细胞活力，抑制炎症反应，并减轻焦亡。RBM15通过依赖IGF2BP2的m6A修饰促进炎性小体NLRP3和NLRC4的表达。
RBM15通过依赖IGF2BP2的m6A修饰促进炎性小体NLRP3和NLRC4的表达，从而促进焦亡。
