艳山姜挥发油调控PPARγ/ABCA1信号抑制巨噬细胞源性泡沫细胞形成的作用机制  

Mechanism of essential oil from Fructus Alpinia zerumbet on inhibiting the formation of macrophage-derived foam cells via PPARγ/ABCA1 signal

作　　者：王声全 张光琼 甘诗泉 肖潮达 徐旖旎[1,2] 陈妍[1,2] 陶玲[1,2] 沈祥春[1,2] WANG Sheng-quan;ZHANG Guang-qiong;GAN Shi-quan;XIAO Chao-da;XU Yi-ni;CHEN Yan;TAO Ling;SHEN Xiang-chun(Key Laboratory of Optimal Utilization of Natural Medicine Resources,School of Pharmaceutic Sciences,Guizhou Medical University,Guiyang 550025,China;The High Efficacy Application of Natural Medicinal Resources Engineering Center of Guizhou Province,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学天然药物资源优效利用重点实验室,贵阳550025 [2]贵州医科大学贵州省特色天然药物高效利用工程中心,贵阳550025

出　　处：《中华中医药杂志》2022年第2期1118-1122,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81760725);国家自然科学基金喀斯特中心项目(No.U1812403-4-4);贵州省科技基金重点项目(No.黔科合基础[2020]1Z069);贵州省科技基金(No.黔科合基础[2019]1279号);贵州省高层次创新型人才百层次人才(No.黔科合人才[2015]4029号);贵州省药学国际科技合作基地(No.黔科合平台人才[2017]5802)。

摘　　要：目的:探讨艳山姜挥发油(EOFAZ)调控PPARγ/ABCA1抑制氧化低密度脂蛋白(ox-LDL)诱导的巨噬细胞源性泡沫细胞形成的作用机制。方法:将THP-1细胞用佛波酯(PMA,100μg/L)诱导分化为巨噬细胞,噻唑蓝(MTT)法检测EOFAZ对巨噬细胞活力的影响。油红O染色法检测脂滴的含量;酶法检测总胆固醇(TC)和游离胆固醇(FC)的含量,并计算胆固醇酯(CE)的含量和占比值;qRT-PCR和Western Blot法分析巨噬细胞中PPARγ、ABCA1 mRNA和蛋白的表达。采用PPARγ的抑制剂GW9662作为反向探针,分析PPARγ、ABCA1 mRNA和蛋白质的表达,以及脂滴累积情况。结果:EOFAZ在0.2~5μg/L对巨噬细胞无明显毒性,EOFAZ低、高剂量(1、5μg/L)可显著减少巨噬细胞内脂滴的累积,TC和CE的含量,以及CE/TC比值(P<0.05,P<0.01),同时上调PPARγ、ABCA1 mRNA和蛋白的表达(P<0.05,P<0.01)。加入GW9662后,PPARγ和ABCA1 mRNA和蛋白的表达减少(P<0.01),脂滴累积增加,逆转了EOFAZ的作用。结论:EOFAZ对ox-LDL诱导的巨噬细胞源性泡沫细胞的形成具有抑制作用,其作用机制与促进PPARγ表达上调密切相关。Objective: To explore the inhibitory effect and potential mechanism of essential oil from Fructus Alpinia zerumbet(EOFAZ) on ox-LDL mediated macrophage-derived foam cell formation in vitro. Methods: THP-1 monocyte was differentiated to macrophage after incubated with 100 μg/L PMA. The macrophage viability of EOFAZ was determined by MTT method and the content of lipid droplets was measured by Oil red O staining method. The contents of total cholesterol and free cholesterol in macrophages were detected by enzymatic method, and the content and ratio of cholesterol ester were calculated.qRT-PCR and Western Blot were used to analyze the mRNA and protein expression of PPARγ, ABCA1 in macrophages.GW9662 as a reversed probe, an inhibitor of PPARγ, was used to detect the mRNA and protein expression of PPARγ, ABCA1 as well as lipid droplet accumulation. Results: EOFAZ had no obvious toxicity to macrophages at the concentration of 0.2~5 μg/L,low and high dose EOFAZ(1, 5 μg/L) could significantly attenuate the accumulation of lipid droplets in macrophages, the content of TC and CE, and the ratio of CE/TC(P<0.05, P<0.01). EOFAZ could up-regulate mRNA and protein of PPARγ and ABCA1(P<0.05, P<0.01). After the addition of GW9662, the mRNA and protein expression of PPARγ and ABCA1 significantly decreased(P<0.01), and the accumulation of lipid droplets increased, which reversed the protection effect of EOFAZ. Conclusion:EOFAZ can inhibit the formation of macrophage-derived foam cells induced by ox-LDL via up-regulation of PPARγ expression.

关 键 词：艳山姜挥发油 氧化低密度脂蛋白 泡沫细胞 过氧化物酶体增殖物激活受体Γ 三磷酸腺苷结合盒转运体A1 动脉粥样硬化 

分 类 号：R285[医药卫生—中药学]