黄芪四君子汤调节T细胞PD1泛素化水平重塑肿瘤免疫微环境抑制胃癌增殖的研究

谭倩影; 谢贵萍; 李响; 刘莲芳; 吴冠楠; 吴晓宇; 翟競

doi:10.14148/j.issn.1672-0482.2023.0629

黄芪四君子汤调节T细胞PD1泛素化水平重塑肿瘤免疫微环境抑制胃癌增殖的研究

doi: 10.14148/j.issn.1672-0482.2023.0629

谭倩影^1,,
谢贵萍¹,
李响¹,
刘莲芳²,
吴冠楠³,
吴晓宇³,
翟競^{1, 3, ,}

1.
南京中医药大学第一临床医学院, 江苏南京 210023
2.
南京中医药大学附属张家港中医院, 江苏苏州 320582
3.
南京中医药大学附属医院/江苏省中医院消化系肿瘤外科, 江苏南京 210029

基金项目:

国家自然科学基金青年科学基金项目 82003180

江苏省自然科学基金青年项目 BK20201094

详细信息

作者简介:
谭倩影, 女, 硕士研究生, E-mail: tqy_1113@163.com

通讯作者:
翟競, 女, 副教授, 副主任医师, 主要从事消化系肿瘤的临床与基础研究, E-mail: zhaij0429@njucm.edu.cn

中图分类号: R285.5
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- 被引次数: 0
出版历程
- 收稿日期: 2022-11-12
- 网络出版日期: 2023-07-15
- 发布日期: 2023-07-10

Huangqi Sijunzi Decoction Restrained Gastric Cancer Proliferation via Regulating the Level of PD1 Ubiquitination on T Cells and Remodeling the Tumor Immune Microenvironment

1.
The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing 210023, China
2.
Zhangjiagang Hospital of Traditional Chinese Medicine, Nanjing University of Chinese Medicine, Suzhou 320582, China
3.
Department of Gastrointestinal Oncology, Jiangsu Province Hospital of Chinese Medicine, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China

摘要

摘要: 目的探讨黄芪四君子汤对MFC胃癌荷瘤小鼠肿瘤增殖及免疫功能的影响。方法建立C57BL/6小鼠MFC胃癌皮下荷瘤模型, 采用随机数字表法, 将24只C57BL/6小鼠分为空白对照组、模型组、中药低剂量组(6.24 g·kg^-1)、中药高剂量组(12.48 g·kg^-1), 每组6只; 记录小鼠体质量, 计算肿瘤体积; ELISA法检测各组小鼠外周血中IFN-γ、IL-2、IL-8和TNF-α表达水平; 应用流式细胞术分别检测小鼠肿瘤、脾脏和胸腺中CD4⁺、CD8⁺、PD1⁺ T细胞亚群的表达水平; 检测小鼠肿瘤中PD1泛素化水平。结果与空白对照组比较, 皮下荷瘤模型组小鼠体质量增加(P < 0.05)，外周血中IFN-γ、IL-2和TNF-α表达水平降低(P < 0.001)、IL-8表达升高(P < 0.05)。与模型组相比, 中药高剂量组小鼠肿瘤大小明显低于模型组(P < 0.05)，外周血中IFN-γ、IL-2和TNF-α表达水平显著上升(P < 0.001，P < 0.01), 而IL-8的表达水平显著降低(P < 0.05);中药高剂量组小鼠的肿瘤、脾脏和胸腺中CD4⁺、CD8⁺T细胞亚群表达均增加(P < 0.001), PD1⁺T细胞则随着其泛素化水平增加(P < 0.01)而表达下降(P < 0.001)。结论黄芪四君子汤可上调MFC荷瘤小鼠CD4⁺、CD8⁺T细胞表达, 调节外周血中细胞因子的表达水平, 并通过增加T细胞胞内泛素化水平, 促使PD1表达下调, 从而抑制胃癌细胞的增殖。
- 胃癌 /
- 黄芪四君子汤 /
- 免疫微环境 /
- 泛素化 /
- PD1
Abstract: OBJECTIVE To evaluate the effects of Huangqi Sijunzi Decoction on tumor proliferation and immune function of MFC gastric cancer bearing mice. METHODS The MFC gastric cancer subcutaneous tumor model of C57BL/6 mouse was established. 24 C57BL/6 mice were divided into control group, model group, low dose Chinese medicine group (6.24 g· kg^-1) and high dose Chinese medicine group (12.48 g· kg^-1), with 6 mice in each group. The body weight and tumor volume of these mice were recorded. Expression levels of IFN-γ, TNF-α, IL-2 and IL-8 in peripheral blood of mice in each group were detected by ELISA. The expression levels of CD4⁺, CD8⁺, and PD1⁺ T cell subsets in the tumor, spleen, and thymus of mice were analyzed by flow cytometry. Detect PD1 ubiquitination levels in mouse tumors. RESULTS Compared with control group, the body weight was increased (P < 0.05), the expression of IFN-γ, TNF-α, IL-2 were decreased (P < 0.001), and the expression of IL-8 was increased in model group (P < 0.05). Compared with model group, the tumor volume of mice in high dose Chinese medicine group was remarkable declined (P < 0.05). The expression levels of IFN-γ, IL-2, TNF-α in mouse peripheral blood increased (P < 0.01, P < 0.001), but the expression level of IL-8 decreased (P < 0.05). The expression of CD4⁺ and CD8⁺ T cell subsets in tumor, spleen, and thymus of mice was increased in high dose Chinese medicine group (P < 0.001). The PD1⁺ T cells were decreased (P < 0.001) with the increase of ubiquitination level (P < 0.01). CONCLUSION Huangqi Sijunzi Decoction can upregulate the expression of CD4⁺and CD8⁺ T cells in MFC tumor bearing mice, regulate the expression level of cytokines in peripheral blood, and promote the downregulation of PD1 expression by increasing the level of intracellular ubiquitination of T cells, thus inhibiting the proliferation of gastric cancer cells.
- gastric cancer /
- Huangqi Sijunzi Decoction /
- immune microenvironment /
- ubiquitination /
- PD1

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图 1 黄芪四君子汤对MFC胃癌荷瘤小鼠体质量的影响

注：与空白对照组比较，^*P < 0.05；与模型组比较，^#P < 0.05。x±s, n=6。

Figure 1. Effect of Huangqi Sijunzi Decoction on the body mass of MFC gastric cancer bearing mice

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图 2 黄芪四君子汤对MFC胃癌荷瘤小鼠肿瘤大小的影响

注: 与模型组比较, ^*P < 0.05, ^**P < 0.01。x±s, n=6。

Figure 2. Effect of Huangqi Sijunzi Decoction on the size of MFC gastric cancer bearing mice

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图 3 黄芪四君子汤对MFC胃癌荷瘤小鼠血清IFN-γ、IL-2、IL-8和TNF-α水平的影响

注: 与空白对照组比较，^#P < 0.05，^###P < 0.001;与模型组比较, ^*P < 0.05，^**P < 0.01，^***P < 0.001。x±s, n=6。

Figure 3. Effect of Huangqi Sijunzi Decoction on serum IFN-γ, IL-2, IL-8 and TNF-α levels of MFC gastric cancer bearing mice

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图 4 黄芪四君子汤对MFC荷瘤小鼠免疫微环境中CD4⁺T细胞的影响

注: A.肿瘤组织; B.脾脏组织; C.胸腺组织; 与模型组比较, ^***P < 0.001。x±s, n=6。

Figure 4. Effect of Huangqi Sijunzi Decoction on CD4⁺T cells in the immune microenvironment of MFC gastric cancer bearing mice

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图 5 黄芪四君子汤对MFC荷瘤小鼠免疫微环境中CD8⁺T细胞的影响

注: A.肿瘤组织; B.脾脏组织; C.胸腺组织; 与模型组比较, ^***P < 0.001。x±s, n=6。

Figure 5. Effect of Huangqi Sijunzi Decoction on CD8⁺ T cells in the immune microenvironment of MFC gastric cancer bearing mice

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图 6 黄芪四君子汤对MFC荷瘤小鼠免疫微环境中PD1⁺T细胞的影响

注: A.肿瘤组织; B.脾脏组织; C.胸腺组织; 与模型组比较, ^***P < 0.001。x±s, n=6。

Figure 6. Effect of Huangqi Sijunzi Decoction on PD1⁺ T cells in the immune microenvironment of MFC gastric cancer bearing mice

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图 7 黄芪四君子汤对MFC荷瘤小鼠肿瘤组织中PD1表达及评分的影响(×200)

注: 与模型组比较, ^**P < 0.01。x±s, n=6。

Figure 7. Effects and immunohistochemical scores of Huangqi Sijunzi Decoction on PD1 expression in tumor tissues of MFC gastric cancerbearing mice (×200)

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图 8 黄芪四君子汤对MFC荷瘤小鼠肿瘤组织中PD1泛素化的影响

注: 与模型组比较, ^**P < 0.01。x±s, n=6。

Figure 8. Effect of Huangqi Sijunzi Decoction on PD1 ubiquitination in the tumor of MFC gastric cancer bearing mice

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