二氢卟吩e6介导的光动力疗法联合免疫检查点抑制剂治疗肺癌的实验研究  

Combination of Photodynamic Therapy and Immune Checkpoint Inhibitors in Treatment of Lung Cancer:An Experimental Study

作　　者：王馨 李扬[2] 郭佳丽 李晓京[2] 冯玫[1,2] WANG Xin;LI Yang;GUO Jiali;LI Xiaojing;FENG Mei(State Key Laboratory of Quantum Optics and Quantum Optics Devices,Institute of Opto-Electronics,Taiyuan,030006,China;The Third Hospital of Shanxi Medical University(Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital))

机构地区：[1]山西大学光电研究所,太原市030006 [2]山西医科大学第三医院·山西白求恩医院·山西医学科学院·同济山西医院

出　　处：《中国激光医学杂志》2024年第1期1-6,58-60,共9页Chinese Journal of Laser Medicine & Surgery

基　　金：国家自然科学基金资助项目(61875113);量子光学与光量子器件国家重点实验室开放课题(KF202002)

摘　　要：目的评估脂质体包裹二氢卟吩e6光动力疗法(Lip@Ce6-PDT)联合免疫检查点抑制剂PD-L1抗体治疗Lewis肺癌模型的疗效。方法(1)采用薄膜蒸发法合成Lip@Ce6,并测定其包封率和载药率。(2)将40只C57BL/6 Lewis肺癌模型小鼠随机分为模型对照组(A组)、PD-L1抗体组(B组)、Lip@Ce6-PDT组(C组)、Lip@Ce6-PDT联合PD-L1抗体组(D组)和正常对照组(E组),共5组,每组8只小鼠。B组小鼠经腹腔内注射PD-L1抗体,剂量为100µg/只;C组小鼠经尾静脉给药1 h后经皮穿刺引入光纤,在肿瘤内部进行激光照射治疗。D组在Lip@Ce6-PDT治疗后腹腔注射与B组等剂量的PD-L1抗体,E组和A组注射等量磷酸盐缓冲液(phosphate buffered solution,PBS),每隔3 d治疗1次,共治疗5次。治疗期间,每日测量肿瘤体积和计算抑瘤率。在第5次治疗结束后3 d,解剖小鼠取肿瘤组织进行HE染色、Bax和Bcl-2免疫组化,通过免疫荧光检测肿瘤组织中CD4^(+)T细胞、CD8+T细胞和Foxp3的浸润以及实时聚合酶链反应(real-time polymerase chain reaction,RT-PCR)检测PD-1和PD-L1 mRNA的表达,并对实验结果进行分析。结果(1)Lip@Ce6的粒径约为103.2 nm,分散性和稳定性良好。Ce6的包封率为68.3%,载药率为8.2%。(2)B组、C组、D组的抑瘤率分别为25.1%、55.4%、68.3%。(3)HE染色结果显示,与A组比较,B组、C组和D组的肿瘤细胞坏死程度依次增加。(4)Bcl-2在肿瘤组织中的阳性表达为A组>B组>C组>D组。Bax在肿瘤组织中的阳性表达为D组>C组>B组>A组。(5)免疫荧光结果显示,相较于A组,B组、C组和D组的CD4+T细胞和CD8+T细胞表达水平依次升高,Foxp3表达水平依次降低。(6)RT-PCR检测结果显示,相较于A组,C组治疗后肿瘤组织中PD-1和PD-L1的mRNA表达量相对增多。结论Lip@Ce6介导的PDT联合免疫检查点抑制剂治疗Lewis肺癌模型具有显著疗效,可有效抑制肿瘤细胞生长。Objective To evaluate the efficacy of liposome-encapsulated dihydroporphyrin e6 photodynamic therapy(Lip@Ce6-PDT)combined with immune checkpoint inhibitor PD-L1 antibody in the treatment of Lewis lung cancer model.Methods Lip@Ce6 was synthesized with membrane evaporation method,and the encapsulation rate and drug loading rate were measured.Forty C57BL/6 Lewis lung cancer model mice were randomly divided into 5 groups:the model control group(group A),PD-L1 antibody group(group B),Lip@Ce6-PDT group(group C),Lip@Ce6-PDT+PD-L1 antibody group(group D)and normal control group(group E),8 mice in each group.PD-L1 antibody was injected intraperitoneally to the mice in group B at a dose of 100μg/mouse.For group C,fiber was introduced into the percutaneous puncture after 1h of tail vein administration,and laser irradiation was then performed inside the tumor.The mice in group D were injected intraperitoneally PD-L1 antibody after Lip@Ce6-PDT treatment.The mice in the blank control group and group A were injected with the same amount of phosphate buffered solution(PBS)once every 3 d for a total of 5 treatments.During the treatment,the tumor volume was measured every day and the tumor inhibitory rate was calculated.Three days after the end of the fifth treatment,the tumor tissue was taken for HE staining,Bax and Bcl-2 immunohistochemistry.Immunofluorescence was used to detect the infiltration of CD4+T cells,CD8+T cells and Foxp3 in tumor tissues,and real-time polymerase chain reaction(RT-PCR)was used to detect the expression of PD-1 and PD-L1 mRNA.The experimental results were analyzed.Results(1)The particle size of Lip@Ce6 was about 103.2 nm and the dispersibility and stability were good.The encapsulation rate of Ce6 was 68.3%,and the drug loading rate was 8.2%.(2)The anti-tumor effect of Lip@Ce6-PDT mediated PDT therapy combined with PD-L1 antibody on mice and the effect on the immune microenvironment.The tumor inhibition rates of group B,C and D were 25.1%,55.4%and 68.3%respectively.(3)The results of HE staining showed th

关 键 词：光动力疗法 免疫检查点抑制剂 联合治疗 肺癌 

分 类 号：R454.2[医药卫生—治疗学] R734.2[医药卫生—临床医学]