苦参碱经TLR4/NF-κB通路对膀胱癌小鼠免疫机制的调控作用  

Effects of matrine on bladder cancer mice through TLR4/NF-κB pathway and its regulation of immunity

作　　者：瞿小祥 孔东波[1] 鲁小红 江波涛[1] 邹伟[1] 查文良[1] QU Xiaoxiang;KONG Dongbo;LU Xiaohong;JIANG Botao;ZOU Wei;ZHA Wenliang(Department of Urology, Xianning Central Hospital/the First Affiliated Hospital of Hubei University of Science and Technology, Xianning 437100, China)

机构地区：[1]咸宁市中心医院/湖北科技学院附属第一医院泌尿外科,湖北咸宁437100

出　　处：《东南大学学报（医学版）》2021年第6期813-819,共7页Journal of Southeast University(Medical Science Edition)

摘　　要：目的:探究苦参碱通过Toll样受体4(TLR4)/核因子(NF)-κB通路对膀胱癌小鼠免疫机制的调控作用。方法:通过皮下注射膀胱癌细胞系T24的方法构建膀胱癌小鼠模型。将36只模型小鼠随机分为对照组、苦参碱低剂量组和苦参碱高剂量组(n=12)。苦参碱腹腔注射干预,剂量分别为5、10 mg·kg^(-1)。测量肿瘤生长情况,采用蛋白质印迹法检测增殖相关蛋白细胞周期蛋白D1(CyclinD1)和增殖细胞核抗原(PCNA)表达,TUNEL染色检测细胞凋亡。比较各组TLR4/NF-κB通路的转录和翻译水平。采用流式细胞术检测各组CD4^(+)、CD8^(+)细胞百分比,免疫组化染色检测肿瘤组织中CD4^(+)细胞浸润情况。结果:3组小鼠各项指标比较差异均有统计学意义(P<0.05)。苦参碱低剂量组和苦参碱高剂量组的肿瘤体积、质量,Cyclin D1和PCNA蛋白表达水平,TLR4、NF-κB mRNA及蛋白表达水平均显著低于对照组,凋亡指数均显著高于对照组(P<0.05);苦参碱高剂量组的肿瘤体积、质量,Cyclin D1和PCNA蛋白表达水平,TLR4、NF-κB mRNA及蛋白表达水平均显著低于苦参碱低剂量组,凋亡指数显著高于苦参碱低剂量组(P<0.05)。3组小鼠的CD4^(+)细胞百分比、CD4^(+)/CD8^(+)和CD4^(+)细胞浸润情况比较差异有统计学意义(P<0.05)。苦参碱低剂量组和苦参碱高剂量组的CD4^(+)细胞百分比、CD4^(+)/CD8^(+)、CD4^(+)细胞浸润水平均显著高于对照组(P<0.05);苦参碱高剂量组的CD4^(+)细胞百分比、CD4^(+)/CD8^(+)、CD4^(+)细胞浸润水平均显著高于苦参碱低剂量组(P<0.05)。结论:苦参碱可能通过调控TLR4/NF-κB途径抑制膀胱癌小鼠模型的肿瘤生长和诱导凋亡,并提高CD4^(+)/CD8^(+)水平促进CD4^(+)细胞的浸润。Objective:To explore the effects of matrine on bladder cancer mice through Toll like receptor 4(TLR4)/nuclear factor(NF)-κB pathway and its regulation of immunity.Methods:A mouse model of bladder cancer was constructed by subcutaneous injection of bladder cancer cell line T24.36 model mice were randomly divided into control group,low-dose matrine group and high-dose matrine group(n=12).Matrine was injected intraperitoneally for intervention,the doses were 5 and 10 mg·kg^(-1).The tumor growth was measured.The proliferation-related proteins CyclinD1 and PCNA were detected by Western blot.TUNEL staining was applied to detect cell apoptosis.The transcription and translation levels of TLR4/NF-κB pathway in each group were compared.The percentage of CD4^(+)and CD8^(+)in each group were detected by flow cytometry.The infiltration of CD4^(+)cells in tumor tissue was detected by immunohistochemical staining.Results:The indicators of the mice in three groups had significant differences(P<0.05).The tumor volume,mass,Cyclin D1 and PCNA protein,TLR4 and NF-κB mRNA and protein levels of the low-dose matrine group and the high-dose matrine group were significantly lower than those of the control group,and the apoptosis index was significantly higher than that of the control group(P<0.05).The tumor volume,mass,Cyclin D1 and PCNA protein,TLR4 and NF-κB mRNA and protein levels of the high-dose matrine group were significantly lower than those of the low-dose matrine group,and the apoptosis index was significantly higher than that of the low-dose matrine group(P<0.05).The CD4^(+)percentage,CD4^(+)/CD8^(+)and CD4^(+)infiltration of the three groups were significantly different(P<0.05).The CD4^(+)percentage,CD4^(+)/CD8^(+)and CD4^(+)infiltration levels of low-dose matrine and high-dose matrine groups were significantly higher than those of the control group(P<0.05).The CD4^(+)percentage,CD4^(+)/CD8^(+)and CD4^(+)infiltration levels of the high-dose matrine group were significantly higher than those of the low-dose matrine group(

关 键 词：膀胱癌 苦参碱 Toll样受体4/核因子-κB途径 免疫抑制 T淋巴细胞 小鼠 

分 类 号：R737.14[医药卫生—肿瘤]