芪玉三龙汤对肺癌小鼠肿瘤组织TLR4/MyD88/NF-κB通路分子表达的影响

Effect of Qiyu Sanlong Decoction on the Molecules Expression of TLR4/MyD88/NF-KB Pathway in the Tumor Tissue of Lung Cancer Mice

目的:研究芪玉三龙汤对肺癌小鼠肿瘤组织TLR4/MyD88/NF-κB通路相关分子表达的影响。方法:基于前期研究基础,采用LLC细胞构建肺癌移植瘤小鼠模型,造模成功后随机分为模型组、顺铂组、芪玉三龙汤20.1 g/kg、40.2 g/kg、80.5 g/kg、芪玉三龙汤80.5 g/kg+顺铂3.67 mg/kg组,每组20只移植瘤小鼠。灌胃相应药物或生理盐水,联合组灌胃及腹腔注射给予,连续治疗21天,剥离瘤组织,称取瘤质量,计算抑瘤率。HE常规染色,光镜下观察肿瘤组织病理形态学。RT-qPCR、Western Blot法检测各组小鼠肿瘤组织中TLR4/MyD88/NF-κB通路相关基因和蛋白表达水平。结果:芪玉三龙汤具有移植瘤抑制作用,量效关系明显,与顺铂联用具有明显协同作用。光镜下观察模型组肿瘤细胞排列紊乱,周围未见坏死肿瘤细胞,而芪玉三龙汤20.1 g/kg、40.2 g/kg、80.5 g/kg组可见不同程度的坏死肿瘤细胞,其中以80.5 g/kg组最为明显。同时该复方能够下调TLR4/MyD88/NF-κB通路中TLR4、MyD88、IRAK、TRAF6、NF-κB及IκB基因和蛋白的表达水平,量效关系明显,且与顺铂联用具有明显协同作用。结论:结合前期研究基础,芪玉三龙汤的抑瘤作用可能与其调控TLR4/MyD88/NF-κB信号通路,改善免疫功能有关。

Objective:To study the effect of Qiyu Sanlong Decoction on the expression of TLR4/MyD88/NF-KB pathway related molecules in lung cancer mice.Methods:Based on previous studies,LLC cells were used to construct transplanted lung cancer mice models.After modeling,the mice were randomly divided into the model group,chemotherapy group(cisplatin),combination group(80.5 g/kg Qiyu Sanlong Decoction+3.67 mg/kg cisplatin)and Qiyu Sanlong Decoction groups(80.48 g/kg、40.24 g/kg、20.12 g/kg),with 20 transplanted tumor mice in each group.Mice were administered orally or intraperitoneally injected with corresponding drugs or saline water,for 21 days constantly.Afterwards,tumor tissues were dissected,the mass of tumors was weighed,and the tumor growth inhibition rate was calculated.HE routine staining was used to observe the pathological morphology of the lung tissue under light microscope.RT-q PCR and Western Blot were used to detect the expression of TLR4/MyD88/NF-KB pathway related molecules and gene in tumor tissues of mice.Results:Qiyu Sanlong Decoction inhibited the growth of transplanted tumors,in significant dose dependant manner.It had a synergistic effect with cisplatin.Under the light microscope,the tumor cells in the model group were disordered and no necrotic tumor cells were observed around them.In Qiyu Sanlong Decoction groups(80.48 g/kg,40.24 g/kg and 20.12 g/kg),different degrees of necrotic tumor cells were observed,and there were the most necrotic tumor cells in 80.48 g/kg group.Meanwhile,Qiyu Sanlong Decoction down-regulated the genes and proteins expression levels of TLR4,MyD88,IRAK,TRAF6,NF-κB and IκB in TLR4/MyD88/NF-κB pathway,in obvious dose-effect relationship,also it showed synergistic effect with cisplatin.Conclusion:Combined with previous studies,this study suggests that the tumor-suppressive effect of Qiyu Sanlong decoction may be related to regulating TLR4/MyD88/NF-κB signaling pathway and improving the immune function.