调衡方对Lewis肺癌荷瘤体巨噬细胞吞噬功能的影响

The Effect of Tiaoheng Prescription on Phagocytic Function of Lewis Lung Cancer Tumor-Bearing Macrophages

目的研究调衡方对Lewis肺癌荷瘤小鼠巨噬细胞形态、数量及活性的影响,探讨调衡方对荷瘤体巨噬细胞作用的免疫调节机制。方法将传代培养的Lewis肺癌细胞接种于小鼠右腋皮下建转移肺癌模型;灌胃给药8 d后,从小鼠腹腔分离抽取巨噬细胞并培养;调衡方各组巨噬细胞培养48 h后,在显微镜下观察活细胞的形态,墨汁吞噬试验以及金葡菌吞噬试验观察巨噬细胞的吞噬能力,细胞酶化学染色观察巨噬细胞胞内酸性磷酸酶的变化,用酶联免疫吸附法(ELISA)检测各培养组M?上清中IL-1、IL-6、TNF-α表达水平。结果调衡方小、中、大剂量各组巨噬细胞培养48 h后,与对照组比较,巨噬细胞体积显著增大,对墨汁和金葡菌的吞噬能力均显著提升,酸性磷酸酶的活性也明显增强,且能抑制炎性因子IL-1、IL-6、TNF-α分泌,这均与给药浓度呈正相关。结论调衡方能增强巨噬细胞的吞噬功能,进而提高胞内酸性磷酸酶等的活性,通过抑制巨噬细胞分泌炎性因子(IL-1、IL-6、TNF-α)有控制巨噬细胞向M2型极化的可能,并减轻肿瘤浸润程度,降低肿瘤发展及转移,而发挥抗肿瘤免疫调节的作用。

Objective To study the effects of Tiaoheng prescription on the morphology,quantity and activity of macrophages in Lewis lung tumor-bearing mice,and to explore the immunoregulatory mechanism of the effect of Tiaoheng prescription on tumor-bearing macrophages.Methods The subcultured Lewis lung cancer cells were inoculated into the right subcutaneous tissue of mice to establish a metastatic lung cancer model.After 8 days of intragastric administration,macrophages were isolated and cultured from the peritoneal cavity of mice.The macrophages of each group were cultured for 48 h.After that,the morphology of living cells was observed under a microscope,the phagocytosis test by ink and the phagocytic test of Staphylococcus aureus were used to observe the phagocytic ability of macrophages,and the changes of intracellular acid phosphatase in macrophages were observed by cell chemical staining.The expression levels of IL-1,IL-6 and TNF-αin the M?supernatant of each culture group were detected by enzymelinked immunosorbent assay(ELISA).Results After 48 hours of culture of macrophages in small,medium and large doses,the volume of macrophages was significantly increased compared with the control group,and the phagocytosis of ink and Staphylococcus aureus was significantly increased.The activity of acid phosphatase was increased significantly,and can inhibit the secretion of inflammatory factors IL-1,IL-6 and TNF-α,which were positively correlated with the concentration of administration.Conclusion Tiaoheng Prescription can enhance the phagocytic function of macrophages,thereby increasing the activity of intracellular acid phosphatase.By inhibiting the secretion of inflammatory factors(IL-1,IL-6,TNF-α)by macrophages,it is possible to regulate the polarization of macrophages to M2,and reduce the degree of tumor infiltration,reduce tumor development and metastasis,and play a role of anti-tumor immune regulation.