The mechanism of SSO regulating SiO2-induced lipid metabolism disorders in macrophages was explored based on lipid metabolomics
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目的 探讨磺基-N-琥珀酰亚胺油酸酯(sulfo-N-succinimidyloleate,SSO)调控二氧化硅(silicon dioxide,SiO2)诱导的巨噬细胞脂质代谢紊乱的机制.方法 于2023年3月,以常规体外培养大鼠肺泡巨噬细胞NR8383,随机分为对照组(C组)、SSO染毒组、SiO2染毒组和SiO2+SSO染毒组,使用SSO和SiO2分别单独或联合染毒NR8383细胞36 h构建细胞模型.免疫荧光和BODIPY 493/503染色分别检测白细胞分化抗原36(cluster of differentiation,CD36)和细胞内脂质的水平,Western blot检测细胞内CD36、肝脏X受体(liver X receptors,LXR)、磷酸化哺乳动物雷帕霉素靶蛋白(P-mammalian target of rapamycin,P-mTOR)、胆碱磷酸转移酶1(cholinephosphotransferase 1,CHPT1)的蛋白表达水平,脂质代谢组学筛选差异脂质代谢物及富集的途径.多组比较采用单因素方差分析,组内两两比较用LSD检验.结果 SiO2染毒导致巨噬细胞CD36、P-mTOR表达增加(P=0.012、0.020),LXR表达降低(P=0.005),细胞内脂质水平升高,给予SSO干预后,与SiO2染毒组比较,SiO2+SSO染毒组巨噬细胞CD36表达降低(P=0.023),LXR表达升高(P=0.000).代谢组学筛选出 C组和SiO2染毒组中有87个差异代谢物,SiO2染毒组和SiO2+SSO染毒组中有19个差异代谢物,两个组中差异代谢物存在5个交集,分别为PS(22∶1/14∶0)、DG(O-16∶0/18∶0/0∶0)、PGP(i-13∶0/i-20∶0)、PC(18∶3/16∶0)、鞘氨酸(SPA).两个比较组差异代谢物均主要富集在甘油磷脂代谢和鞘脂代谢通路.对甘油磷脂代谢通路中的差异基因CHPT1进行验证,SiO2染毒后导致巨噬细胞CHPT1表达降低(P=0.041).结论 SSO可能通过调控PS(22∶1/14∶0)、DG(O-16∶0/18∶0/0∶0)、PGP(i-13∶0/i-20∶0)、PC(18∶3/16∶0)、SPA以及甘油磷脂代谢和鞘脂代谢通路改善SiO2诱导的巨噬细胞脂质代谢紊乱.
Objective To investigate the mechanism of Sulfo-N-succinimidyloleate(SSO)regulating lipid metabolism disorder induced by silicon dioxide(SiO2).Methods In March 2023,Rat alveolar macrophages NR8383 were cultured in vitro and randomly divided into control group(C),SSO exposure group(SSO),SiO2 exposure group(SiO2)and SiO2+SSO exposure group(SiO2+SSO).NR8383 cells were exposure separately or jointly by SSO and SiO2 for 36 h to construct cell models.Immunofluorescence and BODIPY 493/503 staining were used to detect cluster of differentiation(CD36)and intracellular lipid levels,the protein expression levels of CD36,liver X receptors(LXR),P-mammalian target of rapamycin(P-mTOR)and cholinephosphotransferase 1(CHPT1)were detected by Western blot,respectively,and lipid metabolomics was used to screen for different lipid metabolites and enrichment pathways.Single-factor ANOVA was used for multi-group comparison,and LSD test was used for pair-to-group comparison.Results SiO2 caused the expression of CD36 and P-mTOR to increase(P=0.012,0.020),the expression of LXR to decrease(P=0.005),and the intracellular lipid level to increase.After SSO treatment,CD36 expression decreased(P=0.023)and LXR expression increased(P=0.000)in SiO2+SSO exposure group compared with SiO2 exposure group.Metabolomics identified 87 different metabolites in the C group and SiO2 exposure group,19 different metabolites in the SiO2 exposure group and SiO2+SSO group,and 5 overlaps of different metabolites in the two comparison groups,they are PS(22∶1/14∶0),DG(O-16∶0/18∶0/0∶0),PGP(i-13∶0/i-20∶0),PC(18∶3/16∶0),and Sphinganine.In addition,the differential metabolites of the two comparison groups were mainly concentrated in the glycerophospholipid metabolism and sphingolipid metabolism pathways.The differential gene CHPT1 in glycerophospholipid metabolic pathway was verified,and the expression of CHPT1 decreased after SiO2 exposure.Conclusion SSO may improve SiO2-induced lipid metabolism disorders by regulating PS(22∶1/14∶0),DG(O-16∶0/18∶0/0∶0),PGP(i-13∶0/i-20∶0),PC(18∶3/16∶0),SPA,glycerophospholipid metabolism and sphingolipid metabolism pathways.
Lipid metabolomicsCluster of differentiation 36(CD36)Sulfon-succinimiclyloleate(SSO)SilicosisLipid metabolism disorder
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