雷公藤红素调节NOX4信号抑制肝癌相关成纤维细胞转化的机制  

Mechanism of Celastrol Inhibiting the Hepatocellular Carcinoma Associated Fibroblast Transformation by Regulating NOX4 Signal

作　　者：周晓红 盛泳佳 顾艳玲 王瑾 杨毅 韩晨阳 ZHOU Xiao-hong;SHENG Yong-jia;GU Yan-ling;WANG Jin;YANG Yi;HAN Chen-yang(The Second Hospital of Jiaxing,Jiaxing 314001,China)

机构地区：[1]嘉兴市第二医院,浙江嘉兴314001

出　　处：《中药材》2020年第9期2244-2249,共6页Journal of Chinese Medicinal Materials

基　　金：浙江省自然科学基金项目(LYY20H280005,LYY19H280003);浙江省公益技术研究计划(LGF20C090003)。

摘　　要：目的:研究雷公藤红素靶向调节NOX4后抑制肿瘤相关成纤维细胞转化的作用和机制。方法:小鼠成纤维细胞NIH3T3和肝癌H22共培养,将细胞分为对照组及雷公藤红素(5、10、20μmol/L)组。CCK-8法检测成纤维细胞的活力,PI染色后流式细胞术检测细胞周期的改变,免疫荧光染色检测CAFs标志物α-SMA、FAP的表达,蛋白免疫印迹(Western blot)法检测CAFs标志物α-SMA、FAP的表达及NOX4、TGF-β_(1)、Smad_(1)的表达。H_(22)构建荷瘤小鼠模型后,用雷公藤红素进行治疗,免疫组织化学染色检测肿瘤组织中α-SMA、FAP、NOX4、TGF-β_(1)、Smad_(1)表达。结果:雷公藤红素预处理后可抑制与H_(22)细胞共培的NIH3T3的增殖,提高G_(0)/G_(1)期细胞比例,降低G2、S/M期细胞比例,并下调CAFs标志物α-SMA、FAP的表达,抑制NOX4和TGF-β信号。而在H22荷瘤小鼠模型中,雷公藤红素也可显著抑制α-SMA、FAP、NOX4、TGF-β_(1)、Smad_(1)蛋白表达。结论:雷公藤红素可通过NOX4信号抑制肝癌相关成纤维细胞的转化,这是雷公藤红素抗肿瘤的作用机制之一。Objective:To study the effect and mechanism of celastrol on tumor-associated fibroblast transformation after targeted regulation of NOX4.Methods:The fibroblasts NIH3 T3 and hepatocellular carcinoma H_(22) in rats were co-cultured and the cells were divided into control group and celastrol(5,10,20μmol/L)group.CCK-8 method was used to detect the activity of fibroblasts,cell cycle changes were detected by flow cytometry after PI staining,immunofluorescence staining was used to detect the expressions of CAFs markersα-SMA and FAP,Western blot was used to detect the expressions of CAFs markersα-SMA,FAP and TGF-β_(1),NOX4 and Smad1.After H_(22) constructed tumor mice model,mice were treated by celastrol,immunohistochemical staining was used to detect the expressions ofα-SMA and CAFs markers in tumor tissue.Results:Celastrol pretreatment could inhibit the proliferation of NIH3 T3 co-cultured with H_(22) cells,increase the cell proportion in G_(0)/G_(1) period,decrease the cell proportion in G2、S/M periods,down-regulate the expressions of CAFs markersα-SMA and FAP,inhibit the signals of NOX4 and TGF-β.In H_(22) tumor mice model,celastrol also could significantly inhibit the expressions ofα-SMA,FAP,NOX4,TGF-β_(1),Smad1 proteins.Conclusion:Celastrol can inhibit hepatocellular carcinoma associated fibroblast transformation through NOX4 signal,which is one of the anti-tumor mechanisms of celastrol.

关 键 词：雷公藤红素 成纤维细胞 肝癌 NOX4 

分 类 号：R285.5[医药卫生—中药学]