蠲痹历节清方对痛风细胞模型中PPARγ、TLR4、NF-κB的影响  

Effects of Juanbilijieqing Formula on PPARγ, TLR4 and NF-κB In Gout Cell Model

作　　者：郭玉星 熊辉[3] 易法银[3] 朱方晓[3] 陆小龙[4] 周彪[3] 齐新宇[3] 向黎黎 邵先舫 Guo Xingyu;Xiong Hui;Yi Fayin;Zhu Fangxiao;Lu Xiaohu;Zhou Biao;Qi Xinyu;Xiang Lili;Shao Xianfang(Changde First Hospital of traditional Chinese Medicine,Changde,415000,China;Affiliated Hospital of Hunan Academy of traditional Chinese Medicine,Changsha,410006,China;Hunan University of Chinese Medicine,Changsha,410208,China;Hunan University of Chinese Medicine/The Second Affiliated Hospital,Changsha,410005,China)

机构地区：[1]常德市第一中医医院,常德415000 [2]湖南省中医药研究院附属医院,长沙410006 [3]湖南中医药大学,长沙410208 [4]湖南中医药大学第二附属医院,长沙410005

出　　处：《世界科学技术-中医药现代化》2019年第12期2825-2830,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金面上项目(81574005):基于TLR4/NF-κB信号通路的蠲痹历节清方治疗痛风性关节炎机制研究;负责人:熊辉;湖南省研究生科研创新项目(CX2015B317):蠲痹历节清方对TLR4/NF-κB信号通路及PPARγ调控作用的体外实验研究;负责人:郭玉星;湖南省中医药研究院院级重点课题(201807):蠲痹历节清方抗高尿酸血症作用及与尿酸转运蛋白相关性研究,负责人:郭玉星。

摘　　要：目的体外考察蠲痹历节清方抗痛风炎症作用及其分子机制。方法利用单钠尿酸盐(MSU)结晶诱导巨噬细胞建立体外痛风模型,用蠲痹历节清方(44 g·kg^-1)大鼠含药血清(10%),以及PPARγ激动剂吡格列酮20μmol·L^-1、NF-κB阻断剂SN5020μmol·L干预,酶联免疫法测定TNF-α、IL-1β、IL-6含量,RTPCR法与Western-Blot法分别检测PPARγ、TLR4基因转录及蛋白表达,免疫荧光法考察NF-κB核移位。结果与正常对照组比较,MSU诱导巨噬细胞TNF-α、IL-1β、IL-6水平均显著增加(P<0.01),PPARγ基因转录与表达显著降低(P<0.01),TLR4基因转录与表达显著增加(P<0.01),NF-κB核移位。与模型组比较,蠲痹历节清方含药血清,可显著降低TNF-α、IL-1β、IL-6水平(P<0.01),增加PPARγ基因转录与表达(P<0.01),降低TLR4基因转录与表达(P<0.01),阻止NF-κB核移位,其作用与吡格列酮相似。SN50对PPARγ无影响。结伦蠲痹历节清方可能部分通过上调巨噬细胞PPARγ,抑制TLR4/NF-κB信号通路,抑制痛风炎症反应。Objective To investigate the anti-gout inflammation effects of Juanbillijieqing Formula and its molecular mechanism in vitro.Methods Firstly, macrophages were induced by crystallization of monosodium urate(MSU) to establish gout model in vitro. Then, the drug-containing serum(10%) of Juanbillijieqing Formula(44 g·kg^-1) rats, the PPARγ agonists(pioglitazone, 20 μmol·L^-1) and NF-κB blocker(SN50 20 μmol·L-1) were used for intervention. The content of TNF-α, IL-1β and IL-6 was determined by ELISA. The transcriptions and protein expressions of PPARγ and TLR4 genes were detected respectively by RT-PCR and Western-Blot. The nuclear translocation of NF-κB was investigated by immunofluorescence assay. Results Compared with the normal control group, the levels of TNF-α, IL-1β and IL-6 in MSU induced macrophages were increased significantly(P < 0.01);the transcription and expression of PPARγ gene were decreased significantly(P < 0.01);the transcription and expression of TLR4 gene were increased significantly(P < 0.01);and the nucleus of NF-κB was translocated. Compared with the model group, the serum containing Juanbillijieqing Formula could significantly reduce the levels of TNF-α, IL-1β and IL-6(P < 0.01), increase the transcription and expression of PPARγ gene(P < 0.01), decrease the transcription and expression of TLR4 gene(P <0.01), and prevent the nuclear translocation of NF-κB. Its effects were similar to those of pioglitazone. SN50 had no effect on PPARγ. Conclusion Juanbillijieqing Formula may partially inhibit gouty inflammation by up-regulating PPARγ in macrophages and inhibiting TLR4/NF-κB signaling pathway.

关 键 词：蠲痹历节清方 痛风细胞模型 TLR4/NF-κB信号通路 主要元件 作用机制 

分 类 号：R283[医药卫生—中药学]