摘要
目的：探讨消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg细胞免疫平衡的影响。方法：将75只C57BL/6小鼠随机分为消水方（33.5 g·kg（-1））组、顺铂（6 mg·kg（-1））组、消水方（27 g·kg（-1））＋顺铂（6 mg·kg（-1））组、模型组、空白组。除空白组小鼠胸腔内注射磷酸盐缓冲液（PBS）外,其余各组小鼠均采用Lewis肺癌细胞胸腔注射建立模型,连续给药10 d后各组取10只小鼠处死,采用流式细胞技术检测外周血中Th17,Treg细胞数量;流式微球阵列法（CBA）以及酶联免疫吸附法（ELISA）检测外周血中白细胞介素-6（IL-6）,白细胞介素-17A（IL-17A）,白细胞介素-21（IL-21）,白细胞介素-23（IL-23）和转化生长因子-β1（TGF-β1）水平;实时荧光定量聚合酶链式反应（Real-time PCR）,蛋白免疫印迹法（Western blot）检测小鼠脾组织维甲酸相关孤核受体γt（RORγt）,叉状头/翅膀状螺旋转录因子（Foxp3）mRNA及蛋白表达。结果：各组小鼠外周血Th17,Treg细胞数量均高于空白组（P〈0.05,P〈0.01）。其中消水方＋顺铂组、消水方组较模型组能明显增加Th17细胞数量（P〈0.05）,降低Treg细胞数量（P〈0.01）。消水方＋顺铂组IL-6,IL-17A,IL-21含量分别较其余各组均显著升高（P〈0.01）;消水方＋顺铂组较消水方组、模型组IL-23含量升高（P〈0.05）,TGF-β1含量降低（P〈0.05）。与顺铂组、模型组比较,消水方＋顺铂组、消水方组可上调脾组织中RORγt mRNA表达（P〈0.01）,下调Foxp3 mRNA表达（P〈0.05）。与模型组比较,消水方＋顺铂组可上调脾组织RORγt蛋白表达（P〈0.05）,下调Foxp3蛋白表达（P〈0.01）。结论：消水方联合顺铂可能通过上调Th17细胞水平,下调Treg细胞水平,从而纠正Th17/Treg免疫失衡,发挥抑制MPE作用。
Objective： To investigate the effect of Xiaoshui formula（ XSF） combined with cisplatin on Th17/Treg immune balance in lung cancer xenografts in mice with pleural effusion. Method： Seventy five C57 BL/6 mice were randomly divided into XSF group,cisplatin（ DDP） group,Xiaoshui formula ＋ cisplatin（ XSF ＋ DDP）group,model group and blank group. The blank group was injected with phosphate buffered saline（ PBS） buffer via the chest cavity,and the other groups were injected with Lewis lung cancer cells via the chest cavity,in order to establish animal models. After 10 days of continuous administration,10 mice in each group were put to death.The numbers of Th17 and Treg cells in peripheral blood were detected by flow cytometry. The levels of interleukin-6（ IL-6）,interleukin-17 A（ IL-17 A）,interleukin-21（ IL-21）,interleukin-23（ IL-23） and transforming growth factor-β1（ TGF-β1） in peripheral blood were measured by CBA and enzymelinked immuno sorbent assay（ ELISA）. The mRNA and protein expressions of retinoic acid-related orphan receptorγt（ RORγt） and Foxp3 in mouse spleen tissues were detected by Real-time polymerase chain reaction（ PCR） and Western blot. Result： The numbers of Th17 and Treg cells in peripheral blood of all of groups were higher than that in the blank group（ P 0. 05,P 〈 0. 01）. Compared with the model group,the number of Th17 cells was significantly increased（ P 0. 05）,and the number of Treg cells was significantly decreased（ P 〈 0. 01） in both XSF ＋ DDP group and XSF group. The contents of IL-6,IL-17 A and IL-21 in XSF ＋ DDP group were significantly higher than those in other groups（ P 〈 0. 01）. Compared with XSF group,model group and blank group respectively,the XSF ＋ DDP group could significantly increase the content of IL-23（ P 〈 0. 05）,and decrease the content of TGF-β1（ P 〈 0. 05）.Compared with DDP and model group,the expression of RORγt mRNA in spleen tissue was increased in the XSF ＋DDP group and the XSF group（ P 〈 0. 01）,and Foxp3 mRNA expression was reduced（ P 〈 0. 05）. Compared with model group,XSF ＋ DDP group could up-regulate RORγt protein expression（ P 〈 0. 05） and down-regulate Foxp3 protein expression（ P 〈 0. 01） in spleen tissue. Conclusion： Traditional Chinese medicine XSF combined with cisplatin may increase the level of Th17 cells and down-regulate the level of Treg cells,so as to regulate Th17/Treg immune balance and inhibite MPE.
作者
王兵
关江锋
颜琳琳
蔡煜
赵彪
胡作为
侯炜
WANG Bing;GUAN Jiang-feng;YAN Lin-lin;CAI Yu;ZHAO Biao;HU Zuo-wei;HOU Wei(Integrative Medicine Hospital Affiliated of Tongji Medical College,Huazhong University of Science and Technology,Wuhan No.1 Hospital,Wuhan 430022,China;Hanyang Hospital Affiliated of Wuhan University of Science and Technology,Wuhan 430050,china;Sichuan Cancer Hospital,Chengdu 610041,China;Guang＇anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第21期131-137,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81241119)
