摘要: 目的：探索益肺解毒方重塑Lewis肺癌荷瘤鼠免疫抑制微环境的机制。方法：30只C57BL/6小鼠，随机分为5组，分别为空白组、模型组、顺铂(DDP)组、益肺解毒方(YFJDF)组、联合(顺铂 益肺解毒方)组，每组6只。除空白组外，其余组按2.1实验方法建立Lewis肺癌模型，予以相应的药物干预14天，摘眼球取血，处死小鼠，称瘤重，计数肺转移瘤个数，计算抑瘤率和肺转移率；Elase方法检测各组荷瘤鼠及空白组鼠的外周血IL-10、TGF-
β的水平；PT-PCR法检测Lewis肺癌荷瘤鼠肿瘤组织中(Arg-1)，iNOSmRNA表达；流式细胞术检测各组荷瘤鼠肿瘤组织中瘤组织的CD3 、CD4 、CD8 T细胞以及MDSCs、CD4 CD25 T细胞的表达数量。结果：1) DDP组、YFJDF组、DDP YFJDF组荷瘤鼠的瘤重低于Model组，有统计学差异，抑瘤率分别为49.59%、35.98%、68.87%。2) Model组、DDP组、YFJDF组、DDP YFJDF组的荷瘤鼠肺部转移瘤的个数比较无统计学差异，但DDP YFJDF组的肺转移瘤抑制率高于DDP组、YFJDF组。3) 与NC组比较，Model组荷瘤鼠的IL-10、TGF-
β水平明显升高，P < 0.05，有统计学的差异；与Model组比较，DDP组、YFJDF组荷瘤属IL-10、TGF-
β的水平略有降低，但无统计学差异；但DDP YFJDF组荷瘤鼠IL-10、TGF-
β的水平明显降低，P < 0.01，有统计学差异。4) 与Model组比较，DDP组、YFJDF组、DDP YFJDF组的Arg-1mRNA的相对表达量降低，P < 0.01，有统计学差异；与Model组比较，DDP组、YFJDF组、DDP YFJDF组的iNOSmRNA的相对表达量增多，P < 0.01，有统计学差异。5) 与NC组比较，Model组的荷瘤鼠外周血的CD3 、CD4 、CD8 T细胞数目降低，P < 0.01，有显著统计学差异，与Model组比较，DDP组、YFJDF组、DDP YFJDF组荷瘤鼠外周血的CD4 T细胞数目降低，除DDP组外，YFJDF组、DDP YFJDF组的P < 0.01，有显著统计学差异；与Model组比较，DDP组荷瘤鼠外周血的CD8 T细胞数目降低，DDP组的P < 0.05，有统计学差异；YFJDF组、DDP YFJDF组荷瘤鼠外周血的CD8 T细胞数目降低，P < 0.01，有显著统计学差异。6) 与Model组比较，除DDP组外，YFJDF组、DDP YFJDF组的MDSCs的表达显著低于Model组，P < 0.01值有统计学差异；DDP组、YFJDF组、DDP YFJDF组的CD4 CD25 T的表达显著低于Model组，P < 0.01，有统计学差异。结论：Lewis肺癌荷瘤鼠存在免疫抑制，益肺解毒方可作用于MDSCs，降低其分泌的免疫炎性因子，降低了免疫抑制性T细胞的水平，提高了杀伤性T细胞的数目，重塑了免疫抑制的微环境，抑制肺癌生长。
Abstract:
Objective: To explore the mechanism of YFJDF in remodeling the immunosuppressive microenvironment of Lewis lung cancer mice. Methods: Thirty C57BL/6 mice were randomly divided into 5 groups: NC group, Model group, Cisplatin (DDP) group, YFJDF group and (Cisplatin YFJDF) group, with 6 mice in each group. In addition to the blank group, Lewis lung cancer model was established according to the 2.1 experimental method, and corresponding drug intervention was given for 14 days. Blood was taken from eyeballs, mice were killed, a sarcoma was weighed, the number of lung metastases was counted, and anti-tumor rate and lung metastasis rate were calculated. The levels of IL-10 and TGF-β in peripheral blood of each group were detected by Elase. The mRNA expression of Arg-1 and iNOS in Lewis lung cancer tumor tissues was detected by PT-PCR. The expression of CD3 , CD4 , CD8 T cells, MDSCs, CD4 CD25 T cells was detected by flow cytometry. Results: 1) The weight of sarcoma in DDP group, YFJDF group and DDP YFJDF group was lower than that in Model group, and anti-tumor rates were 49.59%, 35.98% and 68.87%, respectively. 2) There was no statistical difference in the number of lung metastases in Model group, DDP group, YFJDF group and DDP YFJDF group, but the inhibition rate of lung metastases in DDP YFJDF group was higher than that in DDP and YFJDF group. 3) Compared with NC group, the levels of IL-10 and TGF-β in Model group were significantly increased (P < 0.05), with statistical difference; Compared with Model group, the levels of IL-10 and TGF-β in DDP group and YFJDF group were slightly decreased, but there was no statistical difference. However, the levels of IL-10 and TGF-β in DDP YFJDF group were significantly decreased (P < 0.01). 4) Compared with Model group, the relative expression of ARG-1 mRNA in DDP group, YFJDF group and DDP YFJDF group decreased (P < 0.01), and the difference was statistically significant. Compared with Model group, the relative expression level of iNOSmRNA in DDP group, YFJDF group and DDP YFJDF group increased (P < 0.01), and the difference was statistically significant. 5) Compared with the NC group, the number of CD3 , CD4 , and CD8 T cells in peripheral blood of Model group decreased (P < 0.01), and the number of CD4 T cells in peripheral blood of Model group, DDP group, YFJDF group, and DDP YFJDF group decreased, except for the DDP group. There was significant difference between YFJDF group and DDP YFJDF group (P < 0.01). Compared with Model group, the number of CD8 T cells in peripheral blood of DDP group decreased, P < 0.05, it was statistically significant. The number of CD8 T cells in peripheral blood of YFJDF group and DDP YFJDF group was significantly different (P < 0.01). 6) Compared with Model group, the expression of MDSCs in YFJDF group and DDP YFJDF group was significantly lower than that in Model group except for DDP group, P < 0.01 it was statistically significant. The expression of CD4 CD25 T in DDP group, YFJDF group and DDP YFJDF group was significantly lower than that in Model group (P < 0.01). Conclusion: Lewis lung cancer tumor bearing mice have immunosuppression, and YFJDF can act on MDSCs, reduce the immune inflammatory factors secreted by MDSCs, reduce the level of immunosuppressive T cells, increase the number of killer T cells, reshape the immunosuppressive microenvironment, and inhibit the growth of lung cancer.