PD-L1抑制促肝源性CD8^(+)IFN-γ^(+)T细胞损伤肝脏功能参与动脉粥样硬化  

PD-L1 inhibits and regulates liver CD8^(+)IFN-γ^(+)T cells to damage liver function and participate in atherosclerosis

作　　者：刘逍 武欣 甄子怡 张嘉滢 李琦[2] 陈畅 LIU Xiao;WU Xin;ZHEN Zi-yi;ZHANG Jia-ying;LI Qi;CHEN Chang(College of Pharmacy,Harbin Medical University,Harbin 150086,China;The Biotherapy Center,Harbin Medical University Cancer Hospital,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学药学院药理学教研室,黑龙江哈尔滨150086 [2]哈尔滨医科大学附属肿瘤医院生物治疗中心,黑龙江哈尔滨150086

出　　处：《中国药理学通报》2025年第4期638-645,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No.81970382);黑龙江省重点研发计划(No.2023ZX06C13)。

摘　　要：目的研究细胞程序性死亡配体-1(programmed cell death protein ligand-1,PD-L1)功能抑制调控免疫活化影响ApoE^(-/-)小鼠动脉粥样硬化发生发展的机制。方法将24只ApoE^(-/-)小鼠随机分为正常组,高脂组和高脂^(+)抗PD-L1单抗组,通过高胆固醇饲料喂养建立动脉粥样硬化(atherosclerosis)模型。实验动物饲养70 d后,分离各组实验动物血管(主动脉根部至腹主动脉)及肝脏组织,进行油红O染色;HE染色检测肝组织病理改变;ELISA检测血清中总胆固醇(CHO)、甘油三酯(TG)、高密度脂蛋白(HDL-c)、低密度脂蛋白(LDL-c)和炎症因子(IFN-γ、TNF-α、IL-1β)含量。流式细胞计数检测肝脏淋巴细胞(CD4^(+)、CD8^(+)、CD4^(+)IFN-γ^(+)和CD8^(+)IFN-γ^(+)T细胞)。RT-PCR检测肝脏组织IFN-γ、TNF-α、IL-1β、CD4和CD8表达。结果与高脂组比较,给予抗PD-L1单抗后促血管壁及肝脏脂质累积并上调血清及肝组织CHO、TG、LDL-c和HDL-c含量。高脂饲养条件下给予抗PD-L1单抗促血清和肝组织谷丙转氨酶(GPT)和谷草转氨酶(GOT)含量升高,但是对碱性磷酸酶(AKP)含量没有影响。高脂饲养条件下给予抗PD-L1单抗促血清和肝脏组织IFN-γ、TNF-α和IL-1β含量升高。高脂饲养条件下给予抗PD-L1单抗抑制CD4表达及促CD8表达。高脂饲养条件下给予抗PD-L1单抗促肝脏CD8^(+)T和CD8^(+)IFN-γ^(+)T细胞活化,但是对CD4^(+)IFN-γ^(+)T细胞活化没有影响。结论高脂饲养条件下给予抗PD-L1单抗通过活化肝脏CD8^(+)IFN-γ^(+)T细胞损伤肝脏功能加重动脉粥样硬化。Aim To study the effect of anti-PD-L1 monoclonal antibody on high-fat diet-induced atherosclerosis in ApoE^(-/-)mice.Methods Twenty-four ApoE^(-/-)mice were randomly divided into the normal group,high-fat group,and high-fat^(+)anti-PD-L1 mAb group.After 70 days,the blood samples were harvested.Blood vessels(aortic root to abdominal aorta)and liver from each groups were stained with Oil Red O.Hematoxylin-eosin staining(HE)was employed to visualize structural changes in liver.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the serum levels of total cholesterol(CHO),triglyceride(TG),high-density lipoprotein(HDL-c),low-density lipoprotein(LDL-c)and inflammatory factors(IFN-γ,TNF-α,IL-1β).Flow cytometry was used to detect the proportion of lymphocytes(CD4 and CD8).RT-PCR was utilized to assess the expressions of IFN-γ,TNF-α,IL-1β,CD4 and CD8 in liver.Results Compared with the high-fat group,the treatment with anti-PD-L1 monoclonal antibody promoted vascular wall and liver lipid accumulation,and also up-regulated serum and liver content of cholesterol(CHO),triglyceride(TG)and high-density lipoprotein(HDL-c).Treatment with anti-PD-L1 monoclonal antibody up-regulated the content of alanine aminotransferase(GPT)and aspartate aminotransferase(GOT)in serum and liver,but not alkaline phosphatase(AKP).ELISA test indicated that treatment with anti-PD-L1 monoclonal antibody stimulated the serum level of IFN-γ,TNF-αand IL-1β.Furthermore,the mRNA level of IFN-γ,TNF-αand IL-1βin liver was also up-regulated after treatment with anti-PD-L1 monoclonal antibody.With flow cytometry,we observed that treatment with anti-PD-L1 monoclonal antibody promoted hepatic CD8^(+)T and CD8^(+)IFN-γ^(+)T cell activation,but had no effect on CD4^(+)IFN-γ^(+)T cell activation under high-fat feeding conditions.Conclusions Anti-PD-L1 monoclonal antibody administered under high-fat feeding conditions can damage liver function and aggravate atherosclerosis by activating liver CD8^(+)IFN-γ^(+)T cells.

关 键 词：动脉粥样硬化 PD-L1 CD8^(+)IFN-γ^(+)T细胞 免疫活化 炎症因子 炎症反应 

分 类 号：R-332[医药卫生] R331.125R392.11R392.12R543.5