白及多糖通过调节免疫作用抑制结肠癌CT26荷瘤小鼠肿瘤生长
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Bletilla Striata polysaccharide inhibits tumor growth in colon cancer CT26 tumor-bearing mice by regulating immunity
LIU Wen-Li，MO Hai-Yun. Oncology Department，Nanhua Hospital Affiliated to Nanhua University，Hengyang 421002，China
刘文立等 白及多糖通过节免疫作用抑制结肠癌 CT26 荷瘤小鼠肿瘤生长 第 8 期
doi：10. 3969/ j. issn. 1000-484X. 2021. 08. 010
白及多糖通过调节免疫作用抑制结肠癌 CT26 荷瘤小鼠肿瘤生长
·941·
刘文立 莫海云① （南华大学附属南华医院肿瘤科，衡阳 421002）
中图分类号 R735. 3+5 文献标志码 A 文章编号 1000-484X（2021）08-0941-05 ［摘 要］ 目的：研究白及多糖（BSP）对结肠癌 CT26 荷瘤小鼠的抗肿瘤及免疫调节作用。方法：从 30 只 BALB/ c 小鼠 中随机选取 6 只作为对照组（Control），剩余 24 只小鼠右前肢皮下接种 CT26 细胞构建 CT26 结肠癌小鼠模型，建模成功后将模 型裸鼠分为荷瘤模型组（Model）、白及多糖低剂量组［BSP-L，100 mg/ ( kg·d）］、白及多糖高剂量组［BSP-H，400 mg/ ( kg·d）］和 阳性药物香菇多糖组［LNT，150 mg/（kg·d）］，每组 6 只。所有小鼠连续灌胃给药 3 周，其中对照组和模型组每天给予与药物组 等量生理盐水灌胃。比较各组肿瘤体积、肿瘤抑制率及免疫器官指数。MTT 法检测脾脏淋巴细胞增殖活性，ELISA 法检测血 清免疫因子 IL-2 和 IFN-γ 水平；流式细胞术检测外周血 T 淋巴细胞亚群；Western blot 检测各组小鼠脾脏组织中 TLR4、My D88、 NF-κB p65 蛋白表达水平。结果：与 Model 组相比，BSP-H 组和 LNT 组移植瘤体积显著缩小，肿瘤重量显著降低，抑瘤率显著增 加（P<0.01）。与 Control 组相比，Model 组小鼠脾脏指数、胸腺指数、脾脏淋巴细胞活性、血清中免疫因子 IL-2 和 IFN-γ 水平、外 周血 CD4+/ CD8+及脾脏组织中 TLR4、MyD88、NF-κB p65 蛋白表达水平显著降低（P<0.05）；与 Model 组相比，BSP-H 组小鼠脾脏 指数、胸腺指数、脾脏淋巴细胞活性、血清中免疫因子 IL-2 和 IFN-γ 水平、外周血 CD4+/ CD8+及脾脏组织中 TLR4、MyD88、NFκB p65 蛋白表达水平显著上升（P<0.05）；而 BSP-L 组与 Model 组相比，以上各指标变化均无显著性差异（P>0.05）。结论：白及 多糖对结肠癌 CT26 荷瘤小鼠肿瘤生长有抑制作用，可能与增强荷瘤小鼠抗肿瘤免疫、激活 TLR4/ NF-κB 信号通路有关。 ［关键词］ CT26 结肠癌；白及多糖；免疫调节；TLR4/ NF-κB 信号通路
［Abstract］ Objective：To study the anti-tumor and immunomodulatory effects of Bletilla Striata polysaccharide（BSP）on CT26 tumor-bearing mice. Methods：6 BALB/ c mice were randomly selected from 30 BALB/ c mice as Control group，and the remaining 24 mice were subcutaneously inoculated with CT26 cells in the right forelimb as CT26 colon cancer mouse model. The 24 tumor bearing nude mice were equally divided into tumor-bearing model group（Model），low-dose BSP group［BSP-L，100 mg/（kg·d）］，high-dose BSP group［BSP-H，400 mg/（kg·d）］and positive group［Lentinan，LNT，150 mg/（kg·d）］. All mice were given intragastrically for 3 weeks，and the control group and model group were given the same amount of saline every day. The tumor volume，tumor inhibition rate and immune organ index were calculated. Proliferation activity of spleen lymphocytes was detected by MTT assay；serum levels of the immune factors like IL-2 and IFN- γ were detected by ELISA；T lymphocyte subsets in peripheral blood were detected by flow cytometry；expression levels of TLR-4，MyD88，and NF- κB p65 were detected by Western blot. Results：Compared with Model group，volumes of transplanted tumor of BSP-H group and LNT group were significantly reduced，weights of tumors were significantly reduced，and the tumor inhibition rate increased significantly（P<0.01）. Compared with Control group，the spleen index，thymus index，spleen lymphocyte activity，serum levels of immune factors IL-2 and IFN- γ，peripheral blood CD4+/ CD8+ ，and TLR4， MyD88，NF-κB p65 protein expression levels in spleen tissues of Model group were significantly decreased（P<0.05）. Compared with Model group，the spleen index，thymus index，spleen lymphocyte activity，serum levels of immune factors IL-2 and IFN-γ，peripher‑ al blood CD4+/ CD8+，and TLR4，MyD88，NF-κB p65 protein expression levels in spleen tissues of BSP-H group were significantly increased（P<0.05）. However，there was no significant difference in changes of the above indicators between BSP-L group and Model group（P>0.05）. Conclusion：Bletilla Striata polysaccharide can inhibit tumor growth in CT26 tumor-bearing mice，which may be relat‑ ed to enhancing tumor-bearing mice anti-tumor immunity and activating TLR4/ NF-κB signaling pathway.
LIU Wen-Li，MO Hai-Yun. Oncology Department，Nanhua Hospital Affiliated to Nanhua University，Hengyang 421002，China
刘文立等 白及多糖通过节免疫作用抑制结肠癌 CT26 荷瘤小鼠肿瘤生长 第 8 期
doi：10. 3969/ j. issn. 1000-484X. 2021. 08. 010
白及多糖通过调节免疫作用抑制结肠癌 CT26 荷瘤小鼠肿瘤生长
·941·
刘文立 莫海云① （南华大学附属南华医院肿瘤科，衡阳 421002）
中图分类号 R735. 3+5 文献标志码 A 文章编号 1000-484X（2021）08-0941-05 ［摘 要］ 目的：研究白及多糖（BSP）对结肠癌 CT26 荷瘤小鼠的抗肿瘤及免疫调节作用。方法：从 30 只 BALB/ c 小鼠 中随机选取 6 只作为对照组（Control），剩余 24 只小鼠右前肢皮下接种 CT26 细胞构建 CT26 结肠癌小鼠模型，建模成功后将模 型裸鼠分为荷瘤模型组（Model）、白及多糖低剂量组［BSP-L，100 mg/ ( kg·d）］、白及多糖高剂量组［BSP-H，400 mg/ ( kg·d）］和 阳性药物香菇多糖组［LNT，150 mg/（kg·d）］，每组 6 只。所有小鼠连续灌胃给药 3 周，其中对照组和模型组每天给予与药物组 等量生理盐水灌胃。比较各组肿瘤体积、肿瘤抑制率及免疫器官指数。MTT 法检测脾脏淋巴细胞增殖活性，ELISA 法检测血 清免疫因子 IL-2 和 IFN-γ 水平；流式细胞术检测外周血 T 淋巴细胞亚群；Western blot 检测各组小鼠脾脏组织中 TLR4、My D88、 NF-κB p65 蛋白表达水平。结果：与 Model 组相比，BSP-H 组和 LNT 组移植瘤体积显著缩小，肿瘤重量显著降低，抑瘤率显著增 加（P<0.01）。与 Control 组相比，Model 组小鼠脾脏指数、胸腺指数、脾脏淋巴细胞活性、血清中免疫因子 IL-2 和 IFN-γ 水平、外 周血 CD4+/ CD8+及脾脏组织中 TLR4、MyD88、NF-κB p65 蛋白表达水平显著降低（P<0.05）；与 Model 组相比，BSP-H 组小鼠脾脏 指数、胸腺指数、脾脏淋巴细胞活性、血清中免疫因子 IL-2 和 IFN-γ 水平、外周血 CD4+/ CD8+及脾脏组织中 TLR4、MyD88、NFκB p65 蛋白表达水平显著上升（P<0.05）；而 BSP-L 组与 Model 组相比，以上各指标变化均无显著性差异（P>0.05）。结论：白及 多糖对结肠癌 CT26 荷瘤小鼠肿瘤生长有抑制作用，可能与增强荷瘤小鼠抗肿瘤免疫、激活 TLR4/ NF-κB 信号通路有关。 ［关键词］ CT26 结肠癌；白及多糖；免疫调节；TLR4/ NF-κB 信号通路
［Abstract］ Objective：To study the anti-tumor and immunomodulatory effects of Bletilla Striata polysaccharide（BSP）on CT26 tumor-bearing mice. Methods：6 BALB/ c mice were randomly selected from 30 BALB/ c mice as Control group，and the remaining 24 mice were subcutaneously inoculated with CT26 cells in the right forelimb as CT26 colon cancer mouse model. The 24 tumor bearing nude mice were equally divided into tumor-bearing model group（Model），low-dose BSP group［BSP-L，100 mg/（kg·d）］，high-dose BSP group［BSP-H，400 mg/（kg·d）］and positive group［Lentinan，LNT，150 mg/（kg·d）］. All mice were given intragastrically for 3 weeks，and the control group and model group were given the same amount of saline every day. The tumor volume，tumor inhibition rate and immune organ index were calculated. Proliferation activity of spleen lymphocytes was detected by MTT assay；serum levels of the immune factors like IL-2 and IFN- γ were detected by ELISA；T lymphocyte subsets in peripheral blood were detected by flow cytometry；expression levels of TLR-4，MyD88，and NF- κB p65 were detected by Western blot. Results：Compared with Model group，volumes of transplanted tumor of BSP-H group and LNT group were significantly reduced，weights of tumors were significantly reduced，and the tumor inhibition rate increased significantly（P<0.01）. Compared with Control group，the spleen index，thymus index，spleen lymphocyte activity，serum levels of immune factors IL-2 and IFN- γ，peripheral blood CD4+/ CD8+ ，and TLR4， MyD88，NF-κB p65 protein expression levels in spleen tissues of Model group were significantly decreased（P<0.05）. Compared with Model group，the spleen index，thymus index，spleen lymphocyte activity，serum levels of immune factors IL-2 and IFN-γ，peripher‑ al blood CD4+/ CD8+，and TLR4，MyD88，NF-κB p65 protein expression levels in spleen tissues of BSP-H group were significantly increased（P<0.05）. However，there was no significant difference in changes of the above indicators between BSP-L group and Model group（P>0.05）. Conclusion：Bletilla Striata polysaccharide can inhibit tumor growth in CT26 tumor-bearing mice，which may be relat‑ ed to enhancing tumor-bearing mice anti-tumor immunity and activating TLR4/ NF-κB signaling pathway.