NADPH氧化酶NOX4抑制剂调节肝癌相关成纤维细胞的形成和机制研究

目的：研究NADPH氧化酶（NOX4）抑制剂GLX351322抑制肿瘤相关成纤维细胞（CAFs）形成的作用和机制。方法：小鼠成纤维细胞NIH3T3和肝癌H22共培养，5 μmol/L（IC50）的GLX351322进行NIH3T3预处理。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和FAP的表达，蛋白免疫印迹（Western blot）法检测CAFs标志物α-SMA、Desmin、FAP、TSP-1、FSP、CyclinD的表达水平以及TGF-β1、Smad1的表达。H22构建荷瘤小鼠模型后，GLX351322处理，免疫组织化学染色检测肿瘤组织中α-SMA的表达情况以及CAFs标志物的表达水平。结果：GLX351322预处理后可以抑制NIH3T3的增殖，细胞活力下调，周期改变。同时可以下调CAFs标志物α-SMA、Desmin、FAP、TSP-1、FSP、CyclinD的表达，且TGF-β信号受到抑制。而在荷瘤小鼠模型中，GLX351322也可以显著抑制α-SMA的表达以及CAFs标志物的表达水平。结论：NOX4抑制剂GLX351322可以抑制肿瘤相关成纤维细胞的形成，其作用和TGF-β信号抑制有关。

NADPH oxidase NOX4 inhibitor regulates the formation and mechanism of hepatoma related fibroblasts

AIM: To study the effect and mechanism of glx351322, an inhibitor of NADPH oxidase (NOX4), on the formation of tumor associated fibroblasts (CAFs). METHODS: NIH3T3 cells were co-cultured with H22 cells. 5 μm (IC50) GLX351322 cells were pretreated with NIH3T3. After PI staining, flow cytometry was used to detect the change of cell cycle, immunofluorescence staining was used to detect the expression of CAFs markers α-SMA and FAP, Western blot was used to detect the expression of CAFs markers α-SMA, Desmin, FAP, TSP-1, FSP, CyclinD, TGF-β1 and Smad1.After H22 was used to construct tumor bearing mice model, GLX351322 was used for treatment,Immunohistochemical staining was used to detect the expression of α-SMA and CAFs in tumor tissue. RESULTS:GLX351322 pretreatment could inhibit the proliferation of NIH3T3, decrease cell viability and change cell cycle. At the same time, it could down-regulate the expression of α-SMA, desmin, FAP, TSP-1, FSP, CyclinD, and TGF-β signal was inhibited. GLX351322 also significantly inhibited the expression of α-SMA and CAFs markers in tumor mice. CONCLUSION: GLX351322, a NOX4 inhibitor, can inhibit the formation of tumor-related fibroblasts, which is related to the inhibition of TGF-β signal.