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目的探讨溶酶体相关膜蛋白3(lysosome associated membrane protein 3,LAMP3)对肝细胞脂质代谢的作用。方法以肝癌细胞系Huh7为研究对象,利用qRT-PCR及Western blot法检测游离脂肪酸(free fatty acids,FFAs)0.4 mmol/L处理后肝细胞LAMP3表达变化;过表达LAMP3后,分别使用油红染色及甘油三酯检测试剂盒检测肝细胞内脂滴及甘油三酯沉积情况;采用ATP及β-羟基丁酸检测试剂盒检测细胞内ATP及β-羟基丁酸含量变化;过表达LAMP3,使用qRT-PCR、Western blot法检测脂质合成相关基因PPARγ与SCD-1及脂质分解相关基因CPT1A与PGC-1α表达变化。结果 FFAs处理后肝细胞Huh7中LAMP3表达显著上调(P<0.05);过表达LAMP3可显著促进肝细胞内脂滴和甘油三酯的沉积(P<0.05);过表达LAMP3对肝细胞内ATP及β-羟基丁酸含量的影响差异无统计学意义(P>0.05);过表达LAMP3可促进PPARγ及SCD-1表达(P<0.05),但对CPT1A、PGC-1α表达的影响差异无统计学意义(P>0.05)。结论 LAMP3可通过调控脂肪合成促进肝细胞内脂质沉积。
Objective To investigate the effect of lysosome associated membrane protein 3 (LAMP3) on lipid metabolism in hepatocytes. Methods The hepatocellular carcinoma cell line Huh7 was used as the research object. The expression of LAMP3 in hepatocytes was detected by qRT-PCR and Western blot after treated with 0.4 mmol / L free fatty acids (FFAs). After overexpression of LAMP3, And triglyceride detection kit detection of intracellular lipid droplets and triglyceride deposition; ATP and beta-hydroxybutyrate detection kit detection of intracellular ATP and beta-hydroxybutyrate content changes; overexpression of LAMP3, the use of qRT The expressions of PPARγ and SCD-1, CPT1A and PGC-1α in lipofection were detected by PCR and Western blot. Results LAMP3 expression was significantly up-regulated in Huh7 cells treated with FFAs (P <0.05). Overexpression of LAMP3 promoted the deposition of lipid droplets and triglycerides in hepatocytes (P <0.05) (P> 0.05). Overexpression of LAMP3 promoted the expression of PPARγ and SCD-1 (P <0.05), but there was no significant difference in the expression of CPT1A and PGC-1α between the two groups Significance (P> 0.05). Conclusion LAMP3 can promote lipid deposition in hepatocytes through regulating fat synthesis.