抑制YAP1提高肠道AKK菌丰度改善PD-1单抗相关肝损伤的机制  

Inhibition of YAP1 increases abundance of intestinal AKK bacteria and improves mechanism of PD-1 monoclonal antibody-related liver damage

作　　者：路峻岚 龚艺 张钰曼 高玉亭 杨彦广 梁华 岳圆 杨佳丽 石新丽 LU Junlan;GONG Yi;ZHANG Yuman;GAO Yuting;YANG Yanguang;LIANG Hua;YUE Yuan;YANG Jiali;SHI Xinli(School of Basic Medical Sciences,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;School of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China)

机构地区：[1]河北中医学院基础医学院,石家庄050200 [2]山西中医药大学基础医学院,晋中030619

出　　处：《中国免疫学杂志》2025年第3期540-544,550,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(82274315);河北省重点研发计划项目卫生健康创新专项(21377735D)。

摘　　要：目的:探讨PD-1单抗治疗时,肝细胞特异性Yap1敲除小鼠提高Akkermansia muciniphila(AKK菌)的丰度并改善肝损伤的机制。方法:选用肝细胞特异性Yap1基因敲除小鼠(基因型为Yap1Flox/Flox;albumin-Cre,标记为Yap1LKO)和对照小鼠(基因型为Yap1Flox/Flox,标记为Yap1Flox),采用马兜铃酸Ⅰ(AAⅠ)与四氯化碳(CCl4)诱导肝损伤模型,将Yap1LKO小鼠和Yap1Flox小鼠随机分为对照组和PD-1单抗组进行干预。干预结束后real-time PCR检测小鼠粪便微生物DNA中AKK菌的丰度变化,16S rRNA基因测序进一步分析肠道菌群变化,HE染色观察小鼠肝脏组织病理学改变。结果:Yap1Flox小鼠出现肝细胞气球样水肿变性、汇管区炎症因子浸润等现象;与Yap1Flox小鼠相比,Yap1LKO小鼠肠内AKK菌丰度降低,肝脏出现纤维化现象,损伤程度更重;单独PD-1单抗治疗不能提高小鼠肠内AKK菌的丰度且肝脏出现纤维增生现象;而PD-1单抗干预的Yap1LKO小鼠肠内AKK菌丰度增加、门水平上厚壁菌门(Firmicutes)与拟杆菌门(Bacteroidetes)的比值(F/B值)降低且肝细胞形态恢复正常、肝损伤程度减轻。结论:AAⅠ+CCl4导致的肝损伤小鼠模型中,肝细胞特异性Yap1敲除降低了肠道内AKK菌丰度,肝损伤程度比Yap1Flox小鼠严重;PD-1单抗治疗时,肝细胞特异性敲除Yap1能提高肠道内AKK菌的丰度、调节肠道菌群组成比例并减轻肝损伤的程度。Objective:To investigate the mechanism of improving liver injury by enhancing the abundance of Akkermansia muciniphila(AKK bacteria)with hepatocyte-specific Yap1 knockout during PD-1 monoclonal antibody treatment.Methods:Hepatocytespecific Yap1 knockout mice(genotype Yap1Flox/Flox;albumin-Cre,labeled as Yap1LKO)and control mice(genotype Yap1Flox/Flox,labeled Yap1Flox),aristolocholic acidⅠ(AAⅠ)and CCl4 were used to induce liver injury,Yap1LKO mice and Yap1Flox mice were randomly divided into control group and PD-1 monoclonal antibody group.After the intervention,real-time PCR detected changes in the abundance of AKK bacteria in the fecal microbial DNA of mice,16S rRNA gene sequencing further analyzes intestinal flora changes,the livers of mice were made into wax blocks for HE staining to observe the histopathological changes of the liver.Results:Yap1Flox mice exhibited balloon-like edema degeneration of hepatocytes and infiltration of inflammatory factors in the manifold area,compared with Yap1Flox mice,Yap1LKO mice had reduced the abundance of AKK bacteria in the intestine,fibrosis of the liver,and the degree of damage was more serious;PD-1 monoclonal antibody treatment alone did not increase the abundance of AKK bacteria in the intestines of mice,and fibrosis appeared in the liver;however,the abundance of intestinal AKK bacteria in Yap1LKO mice treated with PD-1 monoclonal antibody increased,the ratio(F/B value)of Firmicutes to Bacteroides at the phylum level decreased,and the morphology of hepatocytes returned to normal and the degree of liver damage decreased.Conclusion:In AAⅠand CCl4 induced liver injury mice,hepatocyte-specific Yap1 knockout reduced the abundance of AKK bacteria in the intestine,and the degree of liver injury was more severe than that in Yap1Flox mice.When treated with PD-1 monoclonal antibody,hepatocyte-specific Yap1 knockout could increase the abundance of AKK bacteria in the intestine,change the composition of intestinal flora,maintain intestinal homeostasis and ameliorate liver inj

关 键 词：PD-1单抗 YAP1 Akkermansia muciniphila 肝损伤 

分 类 号：R575[医药卫生—消化系统]