吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (6): 1469-1475.doi: 10.13481/j.1671-587X.20210617

• 基础研究 • 上一篇    下一篇

当归六黄汤对前列腺癌荷瘤小鼠的免疫功能调节作用及其机制

胡培森(),崔洪泉,赵俊峰,吴志洲,王交托   

  1. 河南省中医院泌尿外科,河南 郑州 450002
  • 收稿日期:2021-03-06 出版日期:2021-11-28 发布日期:2021-12-14
  • 通讯作者: 胡培森 E-mail:yu589jian@163.com
  • 基金资助:
    河南省中医药管理局中医药科学研究专项课题(2019ZY2043)

Regulatory effect of Danggui Liuhuang Decoction on immune function of prostate cancer-bearing mice and its mechanism

Peisen HU(),Hongquan CUI,Junfeng ZHAO,Zhizhou WU,Jiaotuo WANG   

  1. Department of Urology,Henan University of Traditional Chinese Medicine,Zhengzhou 450002,China
  • Received:2021-03-06 Online:2021-11-28 Published:2021-12-14
  • Contact: Peisen HU E-mail:yu589jian@163.com

摘要: 目的

探讨当归六黄汤对前列腺癌(PCa)荷瘤小鼠免疫功能的调节作用,并阐明其作用机制。

方法

50只小鼠随机分为对照组、模型组、低剂量当归六黄汤组、高剂量当归六黄汤组和顺铂组(n=10)。除对照组外,其他各组小鼠均接种RM-1细胞,建立PCa荷瘤小鼠模型。低和高剂量当归六黄汤组小鼠分别给予0.25和0.50 g·kg-1·d-1当归六黄汤灌胃,顺铂组小鼠腹腔注射0.2 mL顺铂。计算各组小鼠脾脏系数、胸腺系数、肿瘤体积和抑瘤率,HE染色观察各组小鼠肿瘤组织病理形态表现,流式细胞术检测各组小鼠外周血中CD4+和CD8+T淋巴细胞百分率,Western blotting法检测各组小鼠脾脏组织中程序性死亡受体1(PD-1)和程序性死亡配体1(PD-L1)蛋白表达水平。

结果

与对照组比较,模型组小鼠脾脏系数、胸腺系数、CD8+T淋巴细胞百分率、脾脏组织中PD-1和PD-L1蛋白表达水平升高(P<0.05),CD4+T淋巴细胞百分率降低(P<0.05);与模型组比较,低和高剂量当归六黄汤组和顺铂组小鼠脾脏系数、胸腺系数、肿瘤体积、CD8+T淋巴细胞百分率、脾脏组织中PD-1和PD-L1蛋白表达水平降低(P<0.05),CD4+T淋巴细胞百分率升高(P<0.05);与低剂量当归六黄汤组比较,高剂量当归六黄汤组和顺铂组小鼠脾脏系数、胸腺系数、肿瘤体积、CD8+T淋巴细胞百分率、脾脏组织中PD-1和PD-L1蛋白表达水平降低(P<0.05),抑瘤率和CD4+T淋巴细胞百分率升高(P<0.05);与高剂量当归六黄汤组比较,顺铂组小鼠脾脏系数、胸腺系数、肿瘤体积、CD8+T淋巴细胞百分率、脾脏组织中PD-1和PD-L1蛋白表达水平降低(P<0.05),抑瘤率和CD4+T淋巴细胞百分率升高(P<0.05)。HE染色,模型组小鼠肿瘤细胞分布均匀而紧密,偶有空泡;低剂量当归六黄汤组小鼠肿瘤细胞分布均匀且空泡数增多;高剂量当归六黄汤组小鼠肿瘤细胞排列疏松,空泡数明显增多;顺铂组小鼠肿瘤细胞数明显减少,与空泡间隔分布。

结论

当归六黄汤可提高PCa荷瘤小鼠免疫功能,其机制可能与抑制PD-1/PD-L1信号通路有关联。

关键词: 前列腺肿瘤, 当归六黄汤, 免疫调节, 程序性死亡受体1, 程序性死亡配体1

Abstract: Objective

To explore the regulatory effect of Danggui Liuhuang Decoction on the immune function of the prostate cancer (PCa)-bearing mice, and to clarify its mechanism.

Methods

Fifty mice were randomly divided into control group, model group, low dose of Danggui Liuhuang Decoction group, high dose of Danggui Liuhuang Decoction group and cisplatin group (n=10). All the mice were vaccinated with RM-1 to establish the PCa-bearing mouse models except control group.The mice in low dose and high doses of Danggui Liuhuang Decoction groups were given 0.25 and 0.50 g·kg-1·d-1 Danggui Liuhuang Decoction by gavage,respectively,and the mice in cisplatin group were intraperitoneally injected with 0.2 mL cisplatin. The spleen coefficients, thymus coefficients, tumor volumes and inhibitory rates of tumor of the mice in various groups were calculated. HE staining was used to observe the pathomorphology of tumor tissue of the mice in various groups. Flow cytometry was used to detect the percentages of CD4+ and CD8+T lymphocytes in peripheral blood of the mice in various groups. Western blotting method was used to detect the expression levels of programmed cell death receptor-1 (PD-1) and programmed cell death-ligand 1 (PD-L1) proteins in spleen tissue of the mice in various groups.

Results

Compared with control group, the spleen coefficient, thymus coefficient, and percentage of CD8+ T lymphocytes of the mice in model group were increased(P<0.05),and the percentage of CD4+ T lymphocytes was decreased (P<0.05). Compared with model group, the spleen coefficients, thymus coefficients, tumor volumes, percentages of CD8+ T lymphocytes, expression levels of PD-1 and PD-L1 proteins in spleen tissue of the mice in low dose of Danggui Liuhuang Decoction group, high dose of Danggui Liuhuang Decoction group and cisplatin group were decreased(P<0.05),and the percentages of CD4+ T lymphocytes were increased (P<0.05). Compared with low dose of Danggui Liuhuang Decoction group, the spleen coefficients, thymus coefficients, tumor volumes, percentages of CD8+ T lymphocytes, expression levels of PD-1 and PD-L1 proteins in spleen tissue of the mice in high dose of Danggui Liuhuang Decoction group and cisplatin group were decreased (P<0.05), the inhibitory rate of tumor and percentages of CD4+ T lymphocytes were increased (P<0.05).Compared with high dose of Danggui Liuhuang Decoction group, the spleen coefficient, thymus coefficient, tumor volume, percentage of CD8+ T lymphocytes, expression levels of PD-1 and PD-L1 proteins in spleen tissue of the mice in cisplatin group were decreased (P<0.05), and the inhibitory rate of tumor and percentage of CD4+T lymphocytes were increased (P<0.05). The HE staining results showed that the tumor cells in model group were evenly and tightly distributed, with occasional vacuoles,and the tumor cells in low dose of Danggui Liuhuang Decoction group were evenly distributed and the number of vacuoles was increased, the tumor cells in high dose of Danggui Liuhuang Decoction group were loosely arranged and the number of vacuoles was increased significantly, and the tumor cells in cisplatin group were significantly reduced, and they were separated from the vacuole.

Conclusion

Danggui Liuhuang Decoction can improve the immune function of the PCa-bearing mice, and its mechanism maybe related to inhibiting the PD-1/PD-L1 signaling pathway.

Key words: prostate neoplasm, Danggui Liuhuang Decoction, immunomodulation, programmed cell death-1, programmed cell death-ligand1

中图分类号: 

  • R737.25