Zhu Yanji, Li Qian, Chen Yuan, Tian Minle, Xun Wenlong, Sun Shuzhen
Department of Pediatric Nephrology and Rheumatism and Immunology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, P. R. China.
Department of Pediatrics, People's Hospital of Rizhao, Rizhao, P. R. China.
Cell Biol Int. 2022 Mar;46(3):454-461. doi: 10.1002/cbin.11742. Epub 2021 Dec 28.
This study aims to evaluate the effect of purinergic ligand-gated ion channel 7 receptor (P2X7R) antagonist A438079 in kidneys of children with primary nephrotic syndrome (PNS). In vitro, human podocytes were respectively stimulated with oxLDL (80 µg/ml), A438079 (10 µmol/L), or the compound oxLDL and A438079 together. CXC chemokine ligand 16 (CXCL16) and P2X7R expression levels were detected by Western blot and immunofluorescence assay, respectively. Immunofluorescence assay was used to detect Dil-oxLDL, and a Colorimetric Cholesterol Detection Kit was used for quantitative determination. Our results demonstrated that CXCL16 and P2X7R expression levels were remarkably increased in the renal tissue from children with PNS, particularly in the same location. Furthermore, in contrast to children with minimal change disease, the expressions of P2X7R and CXCL16 in renal tissue of children with focal segmental glomerulosclerosis were more obvious. In vitro, CXCL16 and P2X7R expression levels in human podocytes stimulated with oxLDL were markedly elevated accompanying higher intracellular lipid accumulation compared with the normal control group. In addition, pretreatment of human podocytes with A438079 before the start of oxLDL stimulation causes a significant reduction in CXCL16 expression and a decrease in lipid accumulation. Overall, CXCL16 and P2X7R may participate in the progression of PNS. The lipid accumulation reduction caused by A438079 may be through deregulating the CXCL16 pathway, suggesting that there is a potential role for P2X7R antagonists to remedy PNS.
本研究旨在评估嘌呤能配体门控离子通道7受体(P2X7R)拮抗剂A438079对原发性肾病综合征(PNS)患儿肾脏的影响。在体外,分别用氧化型低密度脂蛋白(oxLDL,80μg/ml)、A438079(10μmol/L)或二者联合刺激人足细胞。分别采用蛋白质免疫印迹法和免疫荧光法检测CXC趋化因子配体16(CXCL16)和P2X7R的表达水平。采用免疫荧光法检测Dil-oxLDL,并使用比色法胆固醇检测试剂盒进行定量测定。我们的结果表明,PNS患儿肾组织中CXCL16和P2X7R的表达水平显著升高,特别是在同一部位。此外,与微小病变病患儿相比,局灶节段性肾小球硬化症患儿肾组织中P2X7R和CXCL16的表达更明显。在体外,与正常对照组相比,用oxLDL刺激的人足细胞中CXCL16和P2X7R的表达水平显著升高,同时细胞内脂质积累增加。此外,在oxLDL刺激开始前用A438079预处理人足细胞可导致CXCL16表达显著降低和脂质积累减少。总体而言,CXCL16和P2X7R可能参与PNS的进展。A438079引起的脂质积累减少可能是通过调节CXCL16途径实现的,这表明P2X7R拮抗剂在治疗PNS方面具有潜在作用。