大黄素通过PPARγ/NF-κB信号通路抑制LPS诱导的RAW264.7细胞炎症反应  

Emodin suppresses LPS-induced in ammation in RAW264.7 cells through a PPARγ-dependent pathway

作　　者：任凯旋 涂岳圣 朱涛 骆万婷 林家衍 叶焕荣[1] 方俊杰 

机构地区：[1]南方医科大学临床医学五年制,广东广州510280 [2]南方医科大学珠江医院呼吸内科,广东广州510280

出　　处：《西部医学》2018年第1期12-15,20,共5页Medical Journal of West China

基　　金：中国博士后科学基金(2014M552369);广州市科技计划项目(201707010282);南方医科大学科研启动计划基金(PY2016N015)

摘　　要：目的研究大黄素对LPS诱导的炎症反应的抑制作用及其相关的信号通路。方法将小鼠巨噬细胞RAW264.7细胞分为6组,即Control组、LPS组、大黄素组(Emodin组)、LPS+大黄素组(LPS+Emodin组)、LPS+大黄素组+siRNA-PPARγ(LPS+Emodin+siRNA-PPARγ组)、LPS+大黄素组+siRNA-scrambled(LPS+Emodin+siRNA-scrambled组)。使用ELISA测定TNF-α水平,qPCR和western blot测定ICAM-1、MCP-1和PPARγmRNA和蛋白水平,并使用western blot测定NF-κB p65磷酸化水平。结果与Control组相比较,LPS干预6小时后RAW264.7细胞TNF-α、ICAM-1和MCP-1的表达以及NF-κB p65磷酸化水平均明显升高,PPARγ表达水平明显下降,差异均有统计学意义(P均<0.05);与LPS+Emodin组相比较,LPS组TNF-α、ICAM-1和MCP-1的表达以及NF-κB p65磷酸化水平升高更明显,PPARγ表达水平下降更明显,差异均有统计学意义(P均<0.05);同时siRNA-PPARγ可以有效阻断Emodin的作用,差异均有统计学意义(P均<0.05);且LPS+Emodin组和LPS+Emodin+siRNAscrambled组间各分子表达及磷酸化水平未见明显差别,差异均无明显统计学差异(P>0.05)。结论大黄素可能通过PPARγ/NF-κB信号通路抑制LPS诱导的RAW264.7细胞炎症反应,为大黄素应用于支气管哮喘及ARDS等炎症相关性疾病的临床治疗奠定了基础。Objective To explore whether emodin could attenuate LPS-induced in ammation in RAW264-7 cells, and its involved potential mechanism. Methods RAW264-7 cells were divided into Control group, LPS group, Emodin group, LPS＋Emodin group, LPS＋Emodin＋siRNA-PPARγ and LPS＋Emodin＋siRNA-scrambled group. The mRNA and protein expression of ICAM-1, MCP-1 and PPARγ were measured by qRCR and western blot. The level of TNF-α was evaluated by ELISA. Then, the phosphorylation of NF-κB p65 was also detected by western blot. Meanwhile, siRNA-PPARγ transfection was performed to knockdown PPARγ expression in cells. Results After 6 hours LPS stimulation, LPS-enhanced ICAM-1, MCP-1 and TNF-α expression, LPS-reduced PPARγ expression, and LPS-promoted NF-κB p65 activation were substantially compromised by emdoin in RAW264-7 cells. However, these effects of emodin was noticeably blocked by siRNA-PPARγ in cells. Conclusion Our results indicate that LPS-induced in ammation is potently compromised by emodin very likely through the PPARγ-dependent inactivation of NF-κB in RAW264-7cells.

关 键 词：大黄素 炎症反应 PPARΓ NF—KB RAW264 7细胞 

分 类 号：R285.6[医药卫生—中药学]