金复康口服液对Lewis肺癌荷瘤小鼠PD-1、PD-L1表达及相关免疫因子的影响

Effect of Jinfukang oral liquid on the expression of PD-1,PD-L1 and related immune factors in tumor microenvironment of mice bearing Lewis lung carcinoma

目的:观察金复康口服液对Lewis肺癌小鼠肿瘤微环境中程序性死亡受体1(PD-1)、配体(PD-L1)表达及相关免疫因子的影响。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,而后随机分为模型组、中药低、中、高剂量组每组8只;实验第1天开始中药低、中、高剂量组小鼠分别按15、30、60 g/(kg·d)灌胃给药,模型组按照等量生理盐水给药,1次/d,连续给药14 d;称取各组小鼠的瘤质量,计算抑瘤率;荧光定量PCR检测荷瘤小鼠肿瘤组织中PD-1和PD-L1的mRNA,Western blot法检测肿瘤组织PD-1、PD-L1的蛋白表达水平,HE染色观察肺组织,同时ELISA法检测血清中免疫因子IFN-γ和IL-12表达量。结果:与模型组比较,金复康口服液中、高剂量组能够抑制肺癌移植瘤生长,抑瘤率分别是36.35%和28.59%;金复康口服液中、高剂量组能明显降低肿瘤组织PD-1、PD-L1 mRNA含量,抑制PD-1、PD-L1蛋白表达;HE染色显示中、高剂量组肺组织形态完整,未见明显癌巢;中剂量组明显增加血清中IFN-γ和IL-12的含量。结论:金复康口服液对肺癌移植瘤的生长有一定抑制作用,机制可能与降低肿瘤微环境中PD-1、PD-L1水平并增加血清中免疫因子IFN-γ和IL-12的含量有关。

Objective:To observe the effects of Jinfukang oral liquid(JFK)on the expression of costimulatory molecule programmed receptor 1(PD-1),ligand(PD-L1)and related immune factors in tumor microenvironment of mice bearing Lewis lung carcinoma.Methods:The model of lung cancer subcutaneous allograft was established using Lewis lung cancer cells.After successful tumor implantation,the mice were randomly divided into four groups:A model group and low,medium,and high-dose groups of Chinese medicine,each group containing eight mice.On the first day of the experiment,the low-dose,the medium-dose,and the high-dose of Chinese medicine group were respectively given intragastric administration of 15,30,60 g/(kg·d),and the model group was administered with the same amount of normal saline once daily for 14 days.Tumors of each group were weighed and tumor inhibition rates were calculated.The expressions of PD-1,PD-L1 mRNA and protein in tumor tissues of mice were detected by real-time quantitative PCR and Western blotting,respectively.The lung tissue of mice were observed by HE staining.And IFN-γand IL-12 expression in serum were detected by ELISA.Results:Compared with the model group,the medium and high dose group of JFK could inhibit tumor growth with tumor inhibition rate being 36.35%、28.59%,respectively.The medium and high doses of Jinfukang oral liquid could significantly decrease the expression of PD-1,PD-L1 mRNA and inhibited the expression of the PD-1/PD-L1 protein in tumor tissues;HE staining showed that the lung tissue morphology of the middle and high dose groups was complete,and no obvious cancer nest was observed.The middle dose group significantly increased the levels of IFN-γand IL-12 in serum.Conclusion:Jinfukang oral liquid can inhibit the growth of the transplanted tumor by decreasing PD-1,PD-L1 level in tumor microenvironment and increasing IFN-γand IL-12 levels in serum.