收稿日期: 2023-07-28
网络出版日期: 2024-03-26
基金资助
湖南省卫健委科研计划项目(202202331255)
Involvement of RNF99 in potential link between ubiquitination and septic shock via TAK1/NF⁃κB signaling pathway
Received date: 2023-07-28
Online published: 2024-03-26
目的 探讨环指蛋白99（RNF99）介导的转化生长因子激酶1（TAK1）/核因子-κB（NF-κB）信号通路参与泛素化与脓毒症性急性呼吸窘迫综合征（ARDS）的潜在联系。 方法 进行质粒和siRNA转染以过表达或敲低小鼠肺泡上皮细胞（MLE12）中RNF99，分析磷酸p65和p65蛋白表达。免疫沉淀分析RNF99与TRAF6和TAK1的蛋白相互作用关系。将40只小鼠随机分成WT+PBS、WT+LPS、RNF99特异性表达（TG）+PBS和TG+LPS组，每组10只。通过腹膜内注射30 mg/kg LPS诱导脓毒症。 结果 与Vector组相比，RNF99组MLE12细胞中TRAF6和TAK1的蛋白表达水平显著降低（P＜0.05）。泛素化TRAF6蛋白在RNF99敲低的MLE12细胞中增加。与LPS+Vector组相比，在LPS+RNF99组MLE12细胞中p65的磷酸化水平明显降低（P < 0.05）。与si-NC组相比，si-RNF99组MLE12细胞中RNF99、IκBα的蛋白表达水平显著降低（P < 0.05）。与LPS+si-NC组相比，在LPS+si-RNF99组MLE12细胞中p65的磷酸化水平明显增加（P < 0.05）。TG+LPS组小鼠肺组织中CD68巨噬细胞染色百分比较WT+LPS组显著降低（P < 0.05）。TG+LPS组小鼠肺组织中p65的磷酸化水平显著低于WT+LPS组小鼠（P < 0.05）。 结论 RNF99通过与NF-κB信号通路的关键调节因子（TRAF6/TAK1）相互作用来调节NF-κB信号通路，并改善小鼠腹腔注射LPS后肺损伤。
张弛 , 胡赛 , 王静 , 夏凤强 , 程晓英 , 甘泽英 . RNF99通过TAK1/NF-κB信号通路参与泛素化与脓毒症性休克的潜在联系[J]. 实用医学杂志, 2024 , 40(5) : 615 -620 . DOI: 10.3969/j.issn.1006-5725.2024.05.005
Objective To explore the potential relationship between ubiquitination of transforming growth factor kinase 1 （TAK1）/nuclear factor-κB （NF-κB） signaling pathway mediated by ring finger protein 99 （RNF99） and septic acute respiratory distress syndrome （ARDS）. Methods Plasmid and siRNA transfection were conducted to overexpress or knock down RNF99 in MLE12， and expressions of p65 phosphate and p65 protein were analyzed. The protein interaction between RNF99 and TRAF6 or TAK1 was analyzed by immunoprecipitation assay. Forty mice were randomly divided into WT plus PBS， WT plus LPS， RNF99 specific expression （TG） plus PBS， and TG plus LPS groups， with 10 mice in each group. Sepsis was induced by intraperitoneal injection of 30 mg/kg LPS. Results As compared with vector group， protein expression levels of TRAF6 and TAK1 in MLE12 cells decreased significantly in RNF99 group （P < 0.05）. Ubiquitinated TRAF6 protein increased in MLE12 cells with RNF99 knockdown. As compared with LPS plus vector group， phosphorylation level of p65 in MLE12 cells was significantly lower in LPS plus RNF99 group （P < 0.05）. As compared with si-NC group， protein expression levels of RNF99 and IκBα in si-RNF99 group decreased significantly （P < 0.05）. As compared with LPS plus si-NC group， phosphorylation level of p65 in LPS plus si-RNF99 group increased significantly （P < 0.05）. The staining percentage of CD68 macrophages in lung tissues was significantly lower in TG plus LPS group than in WT plus LPS group （P < 0.05）. Phosphorylation level of p65 in lung tissues was significantly lower in TG plus LPS group than in WT plus LPS group （P < 0.05）. Conclusion RNF99 regulates NF-κB signaling pathway by interacting with the key regulator of NF-κB signaling pathway （TRAF6/TAK1）， and improves lung injury after intraperitoneal injection of LPS in mice.
| 1 | 朱迪， 郭树彬. 脓毒症外周血单个核细胞的免疫特征研究进展［J］. 中华急诊医学杂志， 2022， 31（9）： 1289-1293. doi:10.3760/cma.j.issn.1671-0282.2022.09.026 |
| 2 | 朱永城， 江慧琳， 陈晓辉. 脓毒症休克并发脓毒症心肌病的潜在救治前景：体外膜氧合的挽救性治疗［J］. 中华急诊医学杂志， 2022， 31（7）： 854-857. doi:10.3760/cma.j.issn.1671-0282.2022.07.002 |
| 3 | JING W， WANG H， ZHAN L， et al. Extracellular Vesicles， New Players in Sepsis and Acute Respiratory Distress Syndrome［J］. Front Cell Infect Microbiol， 2022， 12： 853840. doi:10.3389/fcimb.2022.853840 |
| 4 | HUANG H， ZHU J， GU L， et al. TLR7 mediates acute respiratory distress syndrome in sepsis by sensing extracellular miR-146a［J］. Am J Respir Cell Mol Biol， 2022， 67（3）： 375-388. doi:10.1165/rcmb.2021-0551oc |
| 5 | CHEN Y， WU Y， ZHU L， et al. METTL3-mediated N6-methyladenosine modification of Trim59 mRNA protects against sepsis-induced acute respiratory distress syndrome［J］. Front Immunol， 2022， 13： 897487. doi:10.3389/fimmu.2022.897487 |
| 6 | GONG F， LI R， ZHENG X， et al. OLFM4 regulates lung epithelial cell function in sepsis-associated ARDS/ALI via LDHA-mediated NF-κB signaling［J］. J Inflamm Res， 2021， 14： 7035-7051. doi:10.2147/jir.s335915 |
| 7 | SHI X， AN X， YANG L， et al. Reticulocalbin 3 deficiency in alveolar epithelium attenuated LPS-induced ALI via NF-κB signaling［J］. Am J Physiol Lung Cell Mol Physiol， 2021， 320（4）： L627-L639. doi:10.1152/ajplung.00526.2020 |
| 8 | SONG K， LI S. The role of ubiquitination in NF-κB signaling during virus infection［J］. Viruses， 2021， 13（2）： 145. doi:10.3390/v13020145 |
| 9 | ZHANG J， CAO L， GAO A， et al. E3 ligase RNF99 negatively regulates TLR-mediated inflammatory immune response via K48-linked ubiquitination of TAB2［J］. Cell Death Differ， 2023， 30（4）：966-978. doi:10.1038/s41418-023-01115-2 |
| 10 | XIA Q， ZHAN G， MAO M， et al. TRIM45 causes neuronal damage by aggravating microglia-mediated neuroinflammation upon cerebral ischemia and reperfusion injury［J］. Exp Mol Med， 2022， 54（2）： 180-193. doi:10.1038/s12276-022-00734-y |
| 11 | HWANG S J， WANG J H， LEE J S， et al. Ginseng Sprouts Attenuate Mortality and Systemic Inflammation by Modulating TLR4/NF-κB Signaling in an LPS-Induced Mouse Model of Sepsis［J］. Int J Mol Sci， 2023， 24（2）： 1583. doi:10.3390/ijms24021583 |
| 12 | ZHAO J， CAI B， SHAO Z， et al. TRIM26 positively regulates the inflammatory immune response through K11-linked ubiquitination of TAB1［J］. Cell Death Differ， 2021， 28（11）： 3077-3091. doi:10.1038/s41418-021-00803-1 |
| 13 | AKTHER M， HAQUE M E， PARK J， et al. Nlrp3 ubiquitination—A new approach to target nlrp3 inflammasome activation［J］. Int J Mol Sci， 2021， 22（16）： 8780. doi:10.3390/ijms22168780 |
| 14 | WANG W， SHI B， CONG R， et al. RING-finger E3 ligases regulatory network in PI3K/AKT-mediated glucose metabolism［J］. Cell Death Discov， 2022， 8（1）： 372. doi:10.1038/s41420-022-01162-7 |
| 15 | PENG X， WEN Y， ZHA L， et al. TRIM45 suppresses the development of non-small cell lung cancer［J］. Curr Mol Med， 2020， 20（4）： 299-306. doi:10.2174/1566524019666191017143833 |
| 16 | GUO Y， LI Q， ZHAO G， et al. Loss of TRIM31 promotes breast cancer progression through regulating K48-and K63-linked ubiquitination of p53［J］. Cell Death Dis， 2021， 12（10）： 945. doi:10.1038/s41419-021-04208-3 |
| 17 | 彭一鹏， 周晓飞， 王剑， 等. 肿瘤坏死因子α诱导蛋白3相互作用蛋白1调控核因子κB信号通路在IDH野生型胶质瘤中的作用机制［J］. 实用医学杂志， 2022， 38（16）： 2002-2008. doi:10.3969/j.issn.1006-5725.2022.16.004 |
| 18 | ZHANG C， DING Y， LIU Y F， et al. The role of TLR4-mediated MyD88/TRAF6/NF-κB signaling and pIgR intestinal expression in chicks during Salmonella enteritidis infection［J］. Vet Immunol Immunopathol， 2023， 258： 110563. doi:10.1016/j.vetimm.2023.110563 |
| 19 | PARK G D， CHEON Y H， EUN S Y， et al. β-Boswellic acid inhibits RANKL-induced osteoclast differentiation and function by attenuating NF-κB and Btk-PLCγ2 signaling pathways［J］. Molecules， 2021， 26（9）： 2665. doi:10.3390/molecules26092665 |
| 20 | XU C， CHONG L， YU G， et al. MiR-574-5p alleviates sepsis-induced acute lung injury by regulating TRAF6/NF-κB pathway［J］. Tropic J Pharm Res， 2020， 19（4）： 676-682. doi:10.4314/tjpr.v19i4.1 |
| 21 | QIAN J， CHEN X， CHEN X， et al. Kaempferol reduces K63-linked polyubiquitination to inhibit nuclear factor-κB and inflammatory responses in acute lung injury in mice［J］. Toxicol Lett， 2019， 306： 53-60. doi:10.1016/j.toxlet.2019.02.005 |
| 22 | SINGH A K， HAQUE M， O’SULLIVAN K， et al. Suppression of monosodium urate crystal-induced inflammation by inhibiting TGF-β-activated kinase 1-dependent signaling： role of the ubiquitin proteasome system［J］. Cell Mol Immunol， 2021， 18（1）： 162-170. doi:10.1038/s41423-019-0284-3 |
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