浙江医学2025，Vol.47Issue(3)：235-242,8.DOI:10.12056/j.issn.1006-2785.2025.47.3.2024-1100
JMJD3通过TLR4/NF-κB信号通路调控脓毒症诱导的肾小管上皮细胞铁死亡
JMJD3 regulating sepsis-induced ferroptosis of renal tubular epithelial cells via TLR4/NF-κB signaling pathway
摘要
Abstract
Objective To explore the mechanism of Jumonji domain-containing protein 3(JMJD3)regulating ferroptosis of human renal tubular epithelial cells HK-2 induced by sepsis through Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NF-κB)signaling pathway.Methods HK-2 cells were divided into the control group,lipopolysaccharide(LPS)group,LPS+transfection with empty small interfering RNA(siRNA)(si-NC)group,LPS+transfection with JMJD3 siRNA(si-JMJD3)group,LPS+si-JMJD3+ferroptosis inducer(erastin)group,LPS+si-JMJD3+transfection with empty adenovirus(oe-NC)group and LPS+si-JMJD3+transfection with adenovirus overexpressing TLR4(oe-TLR4)group.Cells in the control group were cultured normally,while those in the LPS,LPS+si-NC,LPS+si-JMJD3,LPS+si-JMJD3+erastin,LPS+si-JMJD3+oe-NC,and LPS+si-JMJD3+oe-TLR4 groups were treated with LPS,empty siRNA after LPS treatment,transfection with si-JMJD3 after LPS treatment,LPS and transfection with si-JMJD3 followed by erastin,LPS and transfection with si-JMJD3 followed by transfection with empty adenovirus,and LPS and transfection with si-JMJD3 followed by transfection with adenovirus overexpressing TLR4.Western blot was used to detect the expression levels of JMJD3,glutathione peroxidase 4(Gpx4),ferritin heavy chain 1(FTH-1),TLR4,phosphorylated(p)-NF-κB and NF-κB proteins.Cell counting kit-8 method was used to detect cell activity.Flow cytometry was used to detect cell apoptosis rate.Colorimetry was used to detect the level of Fe2+in cells.Results Compared with the control group,the expression level of JMJD3 protein in LPS group was significantly increased,the cell activity was significantly decreased,and the cell apoptosis rate and the level of Fe2+in cells were significantly increased(all P＜0.05).Compared with LPS+si-NC group,the expression levels of JMJD3,Gpx4,TLR4 and p-NF-κB protein in LPS+si-JMJD3 group were significantly decreased,the expression level of FTH-1 protein was significantly increased,the cell activity was significantly increased,and the cell apoptosis rate and the level of Fe2+in cells were significantly decreased(all P＜0.05).Compared with LPS+si-JMJD3 group,the expression level of Gpx4 protein was significantly increased in LPS+si-JMJD3+erastin group,the expression level of FTH-1 protein was significantly decreased,the cell activity was significantly decreased,and the cell apoptosis rate and the level of Fe2+in cells were both significantly increased(all P＜0.05).Compared with LPS+si-JMJD3+oe-NC group,the expression levels of TLR4 and p-NF-κB protein were significantly increased in LPS+si-JMJD3+oe-TLR4 group,the cell activity was significantly decreased,and the cell apoptosis rate and the level of Fe2+in cells were significantly increased(all P＜0.05).Conclusion JMJD3 can promote sepsis-induced ferroptosis of renal tubular epithelial cells by activating TLR4/NF-κB signaling pathway.关键词
脓毒症/急性肾损伤/Jumonji结构域蛋白3/Toll样受体4/核因子-κB信号通路/铁死亡Key words
Sepsis/Acute kidney injury/Jumonji domain-containing protein 3/Toll-like receptor 4/nuclear factor kappa-B signaling pathway/Ferroptosis引用本文复制引用
金光军,张建成,潘旭鸣,徐步海,陈嘉安,刘佳丽,何煜舟..JMJD3通过TLR4/NF-κB信号通路调控脓毒症诱导的肾小管上皮细胞铁死亡[J].浙江医学,2025,47(3):235-242,8.基金项目
国家中医药管理局科技司-浙江省中医药管理局共建科技计划项目(GZY-ZJ-KJ-24070) （GZY-ZJ-KJ-24070）
浙江省中医药科技计划项目(2022ZA079) （2022ZA079）
