消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg免疫平衡的影响

王兵; 关江锋; 颜琳琳; 蔡煜; 赵彪; 胡作为; 侯炜

doi:10.13422/j.cnki.syfjx.20181924

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消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg免疫平衡的影响

药理 | 更新时间：2024-09-23

- 消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg免疫平衡的影响
- Effect of Xiaoshui Formula Combined with Cisplatin on Th17/Treg Immune Balance in Lung Cancer Xenografts in Mice with Pleural Effusion
- 中国实验方剂学杂志 2018年24卷第21期页码：131-137
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81241119）
- DOI：10.13422/j.cnki.syfjx.20181924
  中图分类号： R-332;R285.5
- 纸质出版日期：2018
- 稿件说明：
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王兵, 关江锋, 颜琳琳, 等. 消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg免疫平衡的影响[J]. 中国实验方剂学杂志, 2018,24(21):131-137.

WANG Bing, GUAN Jiang-feng, YAN Lin-lin, et al. Effect of Xiaoshui Formula Combined with Cisplatin on Th17/Treg Immune Balance in Lung Cancer Xenografts in Mice with Pleural Effusion[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 131-137.
王兵, 关江锋, 颜琳琳, 等. 消水方联合顺铂对肺癌移植瘤胸腔积液小鼠模型Th17/Treg免疫平衡的影响[J]. 中国实验方剂学杂志, 2018,24(21):131-137. DOI： 10.13422/j.cnki.syfjx.20181924.

WANG Bing, GUAN Jiang-feng, YAN Lin-lin, et al. Effect of Xiaoshui Formula Combined with Cisplatin on Th17/Treg Immune Balance in Lung Cancer Xenografts in Mice with Pleural Effusion[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 131-137. DOI： 10.13422/j.cnki.syfjx.20181924.

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。

Abstract

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 C57BL/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-17A (IL-17A)

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-17A 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.

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