PLIN2通过上调ACSL3表达促进THP-1源性巨噬细胞脂质蓄积

PLIN2 promotes lipid accumulation in THP-1 derived macrophages by upregulating ACSL3 expression

摘要:

目的]研究脂肪分化相关蛋白是否通过PI3K/Akt上调酰基辅酶A合成酶长链家族成员3(ACSL3)表达促进巨噬细胞脂质蓄积。 [方法]实验分为24 h组、不同PLIN2表达组、HA-PLIN2＋SC97组、HA-PLIN2＋LY294002组。用Western blot检测细胞中PLIN2、Akt、p-Akt和ACSL3的蛋白表达,RT-qPCR检测细胞内PLIN2 mRNA水平,油红O染色法观察细胞内脂质蓄积程度。 [结果]过表达PLIN2组巨噬细胞中Akt、p-Akt和ACSL3的蛋白表达量显著增高(P＜0.05),p-Akt核移位增加,且PLIN2与Akt的荧光标记重合。在PLIN2过表达的巨噬细胞中加入PI3K/Akt激动剂SC97后,ACSL3的表达量显著增加(P＜0.05),细胞内脂质蓄积程度增加；在PLIN2过表达的巨噬细胞中加入PI3K/Akt抑制剂LY294002后,ACSL3的表达量显著减少(P＜0.05),细胞内脂质蓄积程度减少。 [结论]PLIN2通过PI3K/Akt上调ACSL3表达,从而促进巨噬细胞脂质蓄积。

Abstract:

Aim To investigate whether adipose differentiation-related proteins promote macrophage lipid accumulation by upregulating acyl-CoA synthetase long-chain family member 3 (ACSL3) expression through PI3K/Akt. Methods The experiments were divided into 24 h group, different PLIN2 expression groups, HA-PLIN2＋SC97 group and HA-PLIN2＋LY294002 group. Western blot was used to detect the protein expression of PLIN2, Akt, p-Akt and ACSL3 in cells, RT-qPCR was used to detect the mRNA level of PLIN2 in cells, and oil red O was used to observe the degree of lipid accumulation in cells. Results The protein expression levels of Akt, p-Akt and ACSL3 in macrophages overexpressing PLIN2 were significantly increased (P＜0.05), and p-Akt nuclear translocation was increased, with fluorescence labeling of PLIN2 and Akt overlapping. After adding the PI3K/Akt agonist SC97 to macrophages overexpressing PLIN2, the expression level of ACSL3 significantly increased (P＜0.05), and the degree of intracellular lipid accumulation increased; After adding the PI3K/Akt inhibitor LY294002 to macrophages overexpressing PLIN2, the expression level of ACSL3 significantly decreased (P＜0.05), and the degree of intracellular lipid accumulation decreased. Conclusion PLIN2 upregulates ACSL3 expression through PI3K/Akt, thereby promoting macrophage lipid accumulation.