红芪多糖调控膀胱癌小鼠肿瘤细胞免疫逃逸的研究  

Effects of Astragalus polysaccharides on immune escape of tumor cells in bladder cancer mice

作　　者：樊冬梅 张仕玉 王树文 王冠海 FAN Dong-mei;ZHANG Shi-yu;WANG Shu-wen;WANG Guan-hai(The Second Clinical Medical College,Guangdong Medical University,Dongguan 523808,Guangdong Province,China;Department of Pain,Xianning Ma Tang Hospital of Traditional Chinese Medicine,Xianning 437000,Hubei Province,China;School of Pharmacy,Guangdong Medical University,Dongguan 523808,Guangdong Province,China)

机构地区：[1]广东医科大学第二临床医学院,广东东莞523808 [2]咸宁麻塘中医医院疼痛科,湖北咸宁437000 [3]广东医科大学药学院,广东东莞523808

出　　处：《中国临床药理学杂志》2024年第24期3611-3615,共5页The Chinese Journal of Clinical Pharmacology

基　　金：湖北省卫生健康委员会中医药科研基金资助项目(ZY20210046);广东省基础与应用基础研究基金自然科学基金资助项目(2114050000184)。

摘　　要：目的探讨红芪多糖是否可以通过调控转化生长因子-β1(TGF-β1)/叉头框蛋白P3(FoxP3)/视黄酸受体相关孤儿受体-γt(RORγt)通路来抑制辅助性T细胞17(Th17)、调节性T细胞(Treg)介导的膀胱癌小鼠肿瘤细胞免疫逃逸。方法BALB/c雄性裸鼠臀部注射人膀胱癌细胞系5637细胞构建膀胱癌模型。待造模成功后,随机分为模型组、对照组和低、中、高剂量实验组,每组10只;另取10只正常小鼠作为空白组。低、中、高剂量实验组分别灌胃给予100、200、300 mg·kg^(-1)·d^(-1)的红芪多糖溶液;模型组和空白组均灌胃给予20 mL·kg^(-1)·d^(-1)0.9%NaCl溶液;对照组灌胃给予5 mg·kg^(-1)·d^(-1)顺铂。6组小鼠每天给药1次,连续给药1个月。给药结束后,计算肿瘤抑制率。用TUNEL法检测细胞的凋亡情况,用流式细胞术检测小鼠肿瘤淋巴细胞中Treg和Th7细胞的含量,用蛋白质印迹法检测膀胱组织中TGF-β1、FoxP3和RORγt的表达水平。结果高剂量实验组、对照组和模型组的肿瘤质量分别为(0.89±0.12)、(0.67±0.09)和(2.09±0.31)g,凋亡指数分别为(21.93±2.53)%、(23.91±2.18)%和(4.18±0.39)%;高剂量实验组、对照组、模型组和空白组的Treg细胞含量分别为(3.18±1.15)%、(2.97±0.98)%、(17.23±5.34)%和(2.73±1.04)%,Th7细胞含量分别为(1.03±0.36)%、(0.92±0.33)%、(6.28±1.96)%和(0.86±0.33)%,TGF-β1蛋白相对表达水平分别为0.34±0.13、0.33±0.11、0.69±0.10和0.32±0.08,FoxP3蛋白相对表达水平分别为0.21±0.05、0.20±0.06、0.72±0.16和0.19±0.06,RORγt蛋白相对表达水平分别为0.34±0.12、0.32±0.12、0.66±0.17和0.36±0.09。模型组的上述指标与高剂量实验组和对照组比较,在统计学上差异均有统计学意义(均P<0.05)。结论红芪多糖能够调控TGF-β1/FoxP3/RORγt通路来抑制Treg、Th7细胞介导的膀胱癌小鼠肿瘤细胞免疫逃逸。Objective To explore whether Astragalus polysaccharide can inhibit immune escape of T helper cells 17(Th17)and regulatory T cells(Treg)-mediated tumor cell immune escape in mice with bladder cancer by regulating the transforming growth factor-β1(TGF-β1)/FOXP3/retinoic acid receptor-associated orphan receptor-γT(RORγt)pathway.Methods Human bladder cancer cell line 5637 was injected into the BA LB/c male nude mice to establish the model.After successful mold making,the model mice were randomly divided into model group,control group and experimental-L,-M,-H groups,10 mice per group;another 10 normal mice were taken as the blank group.The experimental-L,-M,-H groups were administered with stragalus polysaccharide solutions at doses of 100,200 and 300 mg·kg^(-1)·d^(-1)by gavage,respectively.The model group and blank group were administered with 0.9%NaCl solution at a dose of 20 mL·k^(-1)·d^(-1)by gavage.The control group was administered with cisplatin at a dose of 5 mg·kg^(-1)·d^(-1)by gavage.Six groups were treated for one month with once a day.After the drug,the tumor suppression was calculated.The cell apoptosis was determined by the TUNEL assay.The counts of Th1and Treg cells were determined by flow cytometry.The expression levels of TGF-β1,FoxP3 and RORγt in bladder tissues were determined by Western blotting.Results The tumor weight of the experimental-H,the control group and the model group were(0.89±0.12),(0.67±0.09)and(2.09±0.31)g,respectively;the apoptosis indexes were(21.93±2.53)%,(23.91±2.18)%and(4.18±0.39)%,respectively.The contents of Treg cells in experimental-H group,control group,model group and blank group were(3.18±1.15)%,(2.97±0.98)%,(17.23±5.34)%and(2.73±1.04)%,respectively;the contents of Th7 cells were(1.03±0.36)%,(0.92±0.33)%,(6.28±1.96)%and(0.86±0.33)%,respectively;the relative expression levels of TGF-β1protein were 0.34±0.13,0.33±0.11,0.69±0.10 and 0.32±0.08,respectively;the relative expression levels of FoxP3 protein were 0.21±0.05,0.20±0.06,0.72±0.16 and

关 键 词：红芪多糖 转化生长因子-β1 叉头框蛋白P3 视黄酸受体相关孤儿受体-γt 免疫逃逸 

分 类 号：R28[医药卫生—中药学]