徐姗, 杜双双, 阎昭. 蚕丝蛋白肽对S180荷瘤小鼠免疫调节作用的研究[J]. 中国肿瘤临床, 2021, 48(1): 1-7. DOI: 10.3969/j.issn.1000-8179.2021.01.089
引用本文: 徐姗, 杜双双, 阎昭. 蚕丝蛋白肽对S180荷瘤小鼠免疫调节作用的研究[J]. 中国肿瘤临床, 2021, 48(1): 1-7. DOI: 10.3969/j.issn.1000-8179.2021.01.089
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Shan Xu, Shuangshuang Du, Zhao Yan. Immunoregulatory effect of silk fibroin peptide in S180 tumor-bearing mice[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2021, 48(1): 1-7. DOI: 10.3969/j.issn.1000-8179.2021.01.089
Citation: Shan Xu, Shuangshuang Du, Zhao Yan. Immunoregulatory effect of silk fibroin peptide in S180 tumor-bearing mice[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2021, 48(1): 1-7. DOI: 10.3969/j.issn.1000-8179.2021.01.089
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蚕丝蛋白肽对S180荷瘤小鼠免疫调节作用的研究

Immunoregulatory effect of silk fibroin peptide in S180 tumor-bearing mice

    摘要
  • 摘要:
      目的  探索蚕丝蛋白肽（silk fibroin peptide，SFP）在肿瘤环境下的免疫调节作用，为临床应用提供药理学基础。
      方法  构建小鼠S180皮下移植瘤模型，分为空白对照组（生理盐水），阳性对照药胸腺五肽组（TP-5，1 mg/kg），蚕丝蛋白肽低剂量组（SFP-L，300 mg/kg）和蚕丝蛋白肽高剂量组（SFP-H，600 mg/kg），腹腔注射连续给药28天。初步评价SFP的抗肿瘤免疫作用；计算免疫脏器指数，评价SFP对于荷瘤小鼠免疫系统发育的影响；苏木精-伊红染色，观察肿瘤组织中的肿瘤浸润性淋巴细胞的数量及分布情况。荷瘤小鼠特异性免疫应答的评价通过流式细胞术测定IL-2、IL-4、IL-5、IFN-γ和TNF-α的分泌水平；MTT法测定T淋巴细胞的增殖能力并计算刺激指数；流式细胞术检测CD4+和CD8+T淋巴细胞亚群的比例；绵羊红细胞免疫小鼠，观察比较小鼠血清的抗体水平。非特异性免疫应答的评价则通过测定小鼠的碳粒廓清指数，衡量巨噬细胞的吞噬能力以及利用MTT法测定自然杀伤细胞的活力。
      结果  SFP具有良好的抗肿瘤活性，可提高胸腺和脾脏的免疫脏器指数（P < 0.05），高剂量SFP可提高肿瘤组织淋巴细胞的浸润程度。在荷瘤小鼠的特异性免疫应答评价中，SFP可提高荷瘤小鼠血清中IL-2、IL-4和TNF-α的分泌水平以及淋巴细胞刺激指数，高剂量SFP可提高T淋巴细胞亚群的比例（P < 0.05）；在非特异性免疫应答评价中，SFP可提高巨噬细胞的吞噬系数α值（P < 0.05）。
      结论  SFP可保护荷瘤小鼠的免疫器官发育，促进肿瘤的局部免疫，还可通过上调特异性及非特异性免疫应答，完善荷瘤小鼠的免疫保护机制，促进荷瘤小鼠的抗肿瘤免疫应答。

     

    Abstract:
      Objective  To investigate the immunoregulatory effects of silk fibroin peptide (SFP) in S180 tumor-bearing mice and provide a solid basis for clinical research into SFP.
      Methods  An S180 subcutaneous tumor mouse model was established, and S180 tumor-bearing mice were divided into four groups: the blank control group received saline, the positive control group received thymopentin (TP-5, 1 mg/kg), and the low- and high-dose SFP groups (SFP-L and SFP-H) received SFP at 300 mg/kg and 600 mg/kg, respectively, via intraperitoneal injection daily for 28 days. Antitumor effects were evaluated first, after which immune response progression was assessed by calculating immune organ (spleen and thymus) indices. Hematoxylin and Eosin staining was used to observe tumor-infiltrating lymphocytes. To evaluate specific immune responses in S180-bearing mice, ratios of CD4+ and CD8+ T-lymphocyte subsets and levels of secreted IL-2, IL-4, IL-5, IFN-γ, and TNF-α were quantified by flow cytometry. An MTT assay was used to assess the proliferative ability of T lymphocytes and calculate their stimulation index; sheep red blood cells were used to immunize mice, and then the serum antibodies were compared. Non-specific immunity was evaluated by carbon particle clearance test, to assess macrophage phagocytic activity, and MTT assay to assess NK cell-mediated cytotoxicity.
      Results  SFP significantly inhibited S180 cell growth in mice and increased immune organ indices (P < 0.05). SFP-H increased lymphocyte infiltration of tumors. SFP stimulated IL-2, IL-4, and TNF-α secretion and increased the lymphocyte stimulation index. SFP-H also increased the ratio of T lymphocyte subsets (P < 0.05), thereby enhancing specific immunity. SFP also increased the macrophage phagocytic coefficient α in our evaluation of non- specific immunity in S180-bearing mice (P < 0.05).
      Conclusions  SFP may protect immune organ development and enhance immune defense systems in tumor microenvironments. Immunity in S180-bearing mice was promoted by upregulating both specific and non-specific immune responses, potentially consolidating these antitumor immunoregulatory effects in a single therapeutic entity.

     

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