α-亚麻酸植物甾醇酯调节SIRT1抑制油酸和胆固醇联合诱导的HepG2细胞焦亡  

PLANT STEROL ESTER OF A-LINOLENIC ACID INHIBITS PYROPTOSIS INDUCED BY OLEIC ACID PLUS CHOLESTEROL BY REGULATING SIRT1 IN HepG2 CELLS

作　　者：李洁 郑明明[2] 裴李圆 田蕾 韩浩[1] LI Jie;ZHENG Ming-ming;PEI Li-yuan;TIAN Lei;HAN Hao(School of Public Health,Shanxi Medical University,Taiyuan 03000l;Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Wuhan 430000,China)

机构地区：[1]山西医科大学公共卫生学院,太原030001 [2]中国农业科学院油料作物研究所,武汉430000

出　　处：《营养学报》2023年第3期264-272,共9页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.81602856);山西省应用基础研究面上青年基金(No.201901D211329);山西省高等学校科技创新基金(No.2019L0420)。

摘　　要：目的 探讨α-亚麻酸植物甾醇酯(PS-ALA)对油酸和胆固醇(OA/Cho)联合诱导的肝细胞焦亡的改善作用,并基于SIRT1信号分子进一步探索潜在的分子机制。方法 采用OA/Cho联合诱导HepG2细胞焦亡,同时给予PS-ALA、PS、ALA干预。油红O染色观察细胞脂质沉积;Hoechst33342/PI双染评价焦亡发生情况;蛋白免疫印迹检测细胞焦亡信号通路关键蛋白NLRP3、Caspase-1 p20、GSDMD-N的表达水平;免疫荧光染色检测NLRP3的表达水平;免疫荧光双标记进行SIRT1和NLRP3以及SIRT1和GSDMD-N共定位。结果 与正常对照组相比,OA/Cho组细胞内有大量脂滴沉积,并且发生明显的细胞焦亡,细胞焦亡信号通路关键蛋白NLRP3、Caspase-1 p20和GSDMD-N的表达水平显著增加,PS-ALA干预后显著改善OA/Cho诱导的细胞内脂质沉积,并有效抑制细胞焦亡。进一步的分子机制研究发现PS-ALA显著上调SIRT1的表达水平并同时降低NLRP3和GSDMD-N的表达水平。结论 PS-ALA能有效抑制OA/Cho诱导的HepG2细胞焦亡,其分子机制可能与激活SIRT1信号分子有关。Objective To investigate the ameliorative effect of plant sterol ester of a-linolenic acid(PS-ALA)on pyroptosis induced by oleic acid and cholesterol(OA/Cho)in HepG2 cells and to further explore the potential molecular mechanism.Methods HepG2 cells were treated by the combination of OA/Cho and intervened by ALA,PS,and PS-ALA.The degree of hepatic cell steatosis was evaluated by oil red O staining.The occurrence of pyroptosis was assessed by Hoechst33342/PI staining.The expression levels of pyroptosis-related proteins(NLRP3,Caspase-1 p20 and GSDMD-N)were detected by Western blot.The colocalization of SIRT1/NLRP3 and SIRT1/GSDMD-N was assessed by double-label immunofluorescence assay.Results Compared with the control group,the OA/Cho group showed significant lipid deposition,obvious pyroptosis and increased expression levels of pyroptosis-related proteins.PS-ALA treatment significantly inhibited OA/Cho-induced lipid deposition and pyroptosis.Further molecular studies showed that PS-ALA significantly up-regulated the protein expression level of SIRT1 and down-regulated NLRP3 the GSDMD-N.Conclusion PS-ALA can effectively inhibit OA/Cho-induced pyroptosis in HepG2 cells,and its molecular mechanism may be related to the up-regulated SIRT1.

关 键 词：α-亚麻酸植物甾醇酯 非酒精性脂肪性肝炎 细胞焦亡 SIRT1 NLRP3 

分 类 号：R151.2[医药卫生—营养与食品卫生学]