目的 探讨复方阿胶浆（FFEJ）改善肺癌肿瘤免疫微环境（TIME）增强程序性死亡蛋白-1（PD-1）抑制剂抗小鼠肺癌的作用。方法 42只C57BL/6J小鼠随机分为对照组，模型组，PD-1抗体（每只200μg）组，FFEJ低、高剂量(10、20 mL·kg^-1)组，PD-1抗体+FFEJ低、高剂量(10、20 mL·kg^-1)组，每组各6只。收集处于对数生长期的Lewis细胞，调节至5×10⁶个·mL^-1，除对照组外，其余组小鼠右腋sc 100μL细胞悬液构建荷瘤小鼠模型。ig给药，体积为20 mL·kg^-1，造模第1天开始，每天1次，持续23 d；对照组和模型组每天ig等体积纯水。PD-1抗体组ip抗PD-1单抗，共5次，每3天1次（第9、12、15、18、21天）。记录小鼠肿瘤体积变化情况；称量小鼠肿瘤质量；苏木精-伊红（HE）染色观察肿瘤组织病理情况；免疫组织化学检测肿瘤组织中增殖细胞相关抗原（Ki67）表达；血常规检测小鼠外周血淋巴细胞（Lym）、粒细胞（Neu）含量及粒细胞/淋巴细胞比值（NLR）；免疫荧光检测肿瘤组织中CD8⁺T细胞和γ干扰素（IFN-γ）表达情况。结果 与模型组比较，单药组和联合给药组小鼠肿瘤体积均显著下降（P<0.001）;FFEJ、PD-1抗体单药组和联合给药高剂量组肿瘤质量均有下降趋势（P>0.05），联合给药低剂量组平均肿瘤质量显著下降（P<0.05）;HE染色显示，FFEJ或PD-1抗体单用对肿瘤细胞的抑制作用不明显，与PD-1抗体联用时肿瘤组织可见大面积坏死区域，显著抑制肿瘤细胞生长，坏死率显著增加（P<0.01、0.001）;FFEJ和PD-1抗体单药组Ki67表达均有下降趋势（P>0.05），联合给药组Ki67表达显著下降（P<0.05）;FFEJ高剂量组和联合给药组Neu、NLR数量显著降低（P<0.05、0.001）,Lym数量显著升高（P<0.001）;FFEJ单药组CD8⁺T、IFN-γ荧光强度并无明显变化，PD-1抗体组IFN-γ荧光强度有升高趋势（P>0.05），联合给药低剂量组CD8⁺T荧光强度有升高趋势（P>0.05）,IFN-γ荧光强度显著升高（P<0.05），联合给药高剂量组CD8⁺T、IFN-γ荧光强度显著升高（P<0.01）。结论 FFEJ可有效改善肺癌TIME，减轻炎症反应，激活机体对肿瘤的免疫应答，增强PD-1抑制剂抗小鼠肺癌的作用。

Objective To explore the effect of Fufang Ejiao Jiang（FFEJ） in improving the tumor immune microenvironment（TIME） of lung cancer and enhancing the anti-lung cancer effect of programmed death protein-1（PD-1） inhibitors in mice. Methods Fortytwo C57 BL/6J mice were randomly divided into control group, model group, PD-1 antibody（200 μg per mouse） group, low and highdose FFEJ(10 and 20 mL·kg^-1) groups, and PD-1 antibody + low and high-dose FFEJ(10 and 20 mL·kg^-1) groups, with 6 mice in each group. Lewis cells in the logarithmic growth phase were collected and adjusted to 5×10⁶ cells·mL^-1. Except for the control group, the remaining groups were subcutaneously injected with 100 μL of cell suspension in the right axilla to establish tumor-bearing mouse models. The mice were administered ig with a volume of 20 mL·kg^-1, starting from the first day of modeling, once a day for 23 days; The control group and the model group were given the same volume of pure water ig every day. The PD-1 antibody group was ip injected with anti-PD-1 monoclonal antibody for a total of 5 times, once every 3 d（on the 9 th, 12 th, 15 th, 18 th, and 21 st d）. The changes in tumor volume of the mice were recorded; The tumor mass in mice was weighed; the pathological conditions of the tumor tissue were observed by hematoxylin-eosin（HE） staining; The expression of proliferation cell-associated antigen（Ki67） in the tumor tissue was detected by immunohistochemistry; The peripheral blood lymphocytes（Lym）, granulocytes（Neu） content and granulocyte/lymphocyte ratio（NLR） of the mice were detected by blood routine; the expression of CD8⁺ T cells and interferon γ（IFN-γ） in the tumor tissue was detected by immunofluorescence. Results Compared with the model group, the tumor volume of the single drug group and the combination drug group decreased significantly（P ＜ 0.001）; The tumor mass of the FFEJ, PD-1 antibody single drug group and the high-dose combination drug group showed a decreasing trend（P＞ 0.05）, and the average tumor mass of the lowdose combination drug group decreased significantly（P ＜ 0.05）; HE staining showed that the inhibitory effect of FFEJ or PD-1 antibody alone on tumor cells was not obvious, but when combined with PD-1 antibody, large areas of necrotic regions were observed in the tumor tissue, significantly inhibiting the growth of tumor cells, and the necrosis rate increased significantly（P ＜ 0.01, 0.001）; The expression of Ki67 in the FFEJ and PD-1 antibody single drug groups showed a decreasing trend（P＞ 0.05）, and the expression of Ki67 in the combination drug group decreased significantly（P ＜ 0.05）; the number of Neu and NLR in the high-dose FFEJ group and the combination drug group decreased significantly（P ＜ 0.05, 0.001）, and the number of Lym increased significantly（P ＜ 0.001）; The fluorescence intensity of CD8⁺ T and IFN-γ in the FFEJ single drug group did not change significantly, the fluorescence intensity of IFN-γ in the PD-1 antibody group showed an increasing trend（P＞ 0.05）, the fluorescence intensity of CD8⁺ T in the low-dose combination drug group showed an increasing trend（P＞ 0.05）, and the fluorescence intensity of IFN-γ increased significantly（P ＜ 0.05）, and the fluorescence intensity of CD8⁺ T and IFN-γ in the high-dose combination drug group increased significantly（P ＜ 0.01）. Conclusion FFEJ can effectively improve TIME of lung cancer, reduce inflammatory responses, activate the immune response of the body to tumors, and enhance the anti-lung cancer effect of PD-1 inhibitors in mice.

R285.5

广东省药品监督管理局科技创新项目（2023ZDZ03）