抑制谷氨酰胺代谢减轻血管紧张素Ⅱ诱导的心肌纤维化  

Inhibition of glutaminolysis alleviates myocardial fibrosis induced by angiotensinⅡ

作　　者：王盼盼 白浩淼 何思宇 夏子淇 刘美杰 安冏 周嘉恒 李宸涵 张伟 张星[2] 王馨佩[2] 李嘉 WANG Pan-Pan;BAI Hao-Miao;HE Si-Yu;XIA Zi-Qi;LIU Mei-Jie;AN Jiong;ZHOU Jia-Heng;LI Chen-Han;ZHANG Wei;ZHANG Xing;WANG Xin-Pei;LI Jia(College of Life Sciences,Northwest University,Xi’an 710069,China;School of Aerospace Medicine,Air Force Medical University;Cadet Regiment,School of Basic Medical Sciences,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]西北大学生命科学学院,西安710069 [2]中国人民解放军空军军医大学航空航天医学系 [3]中国人民解放军空军军医大学基础医学院学员队,西安710032

出　　处：《生理学报》2023年第2期179-187,共9页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China(No.32271150,32071108);the Foundation for Distinguished Young Scholars of Shaanxi Province,China(No.2022JC-15);the Air Force Medical University Talent Support Project(No.2019 RCFCLJ,2023-YBSF-535)。

摘　　要：本文旨在探讨心脏成纤维细胞(cardiac fibroblasts,CFs)谷氨酰胺(glutamine,Gln)代谢在高血压所致心肌纤维化中的作用及机制。用微渗透泵给予C57BL/6J小鼠血管紧张素Ⅱ(angiotensinⅡ,AngⅡ,1.6 mg/kg per d)诱导心肌纤维化,用免疫组织化学法和Western blot检测心肌组织谷氨酰胺酶1(glutaminase 1,GLS1)的表达。小鼠腹腔注射GLS1抑制剂BPTES(12.5 mg/kg)以抑制Gln代谢,用Masson染色法观察心肌纤维化程度,用RT-qPCR和Western blot检测心肌组织Ⅰ型和Ⅲ型胶原蛋白表达变化。Sprague-Dawley(SD)大鼠乳鼠CFs在有/无AngⅡ(0.4μmol/L)刺激下接受Gln 4 mmol/L或BPTES(5μmol/L)处理。用划痕实验和CCK-8法分别检测CFs迁移和增殖,用RT-q PCR和Western blot检测CFs中GLS1、Ⅰ型和Ⅲ型胶原蛋白表达变化。在AngⅡ和BPTES处理的条件下,给予CFs 2 mmol/Lα-酮戊二酸(α-ketoglutarate,α-KG)处理,检测CFs迁移、增殖、Ⅰ型和Ⅲ型胶原蛋白表达变化。结果显示,AngⅡ诱导的小鼠血压、心脏质量和心肌纤维化均增加,同时心肌组织GLS1表达显著上调。在体外实验中,无论有/无AngⅡ刺激,Gln均可显著促进CFs增殖、迁移及GLS1、Ⅰ型和Ⅲ型胶原蛋白的表达,而BPTES显著降低CFs上述指标;补充α-KG可逆转BPTES对AngⅡ刺激下CFs的抑制作用。给小鼠腹腔注射BPTES可改善AngⅡ诱导的小鼠心肌纤维化。以上结果提示,Gln的分解代谢在AngⅡ所致心肌纤维化进程中发挥重要作用,靶向抑制Gln分解代谢可能是治疗心肌纤维化的新策略。The present study was aimed to investigate the role and mechanism of glutaminolysis of cardiac fibroblasts(CFs)in hypertension-induced myocardial fibrosis.C57BL/6J mice were administered with a chronic infusion of angiotensin II(Ang II,1.6 mg/kg per d)with a micro-osmotic pump to induce myocardial fibrosis.Masson staining was used to evaluate myocardial fibrosis.The mice were intraperitoneally injected with BPTES(12.5 mg/kg),a glutaminase 1(GLS1)-specific inhibitor,to inhibit glutaminolysis simultaneously.Immunohistochemistry and Western blot were used to detect protein expression levels of GLS1,Collagen I and Collagen III in cardiac tissue.Neonatal Sprague-Dawley(SD)rat CFs were treated with 4 mmol/L glutamine(Gln)or BPTES(5μmol/L)with or without Ang II(0.4μmol/L)stimulation.The CFs were also treated with 2 mmol/Lα-ketoglutarate(α-KG)under the stimulation of Ang II and BPTES.Wound healing test and CCK-8 were used to detect CFs migration and proliferation respectively.RT-qPCR and Western blot were used to detect mRNA and protein expression levels of GLS1,Collagen I and Collagen III.The results showed that blood pressure,heart weight and myocardial fibrosis were increased in Ang II-treated mice,and GLS1 expression in cardiac tissue was also significantly up-regulated.Gln significantly promoted the proliferation,migration,mRNA and protein expression of GLS1,Collagen I and Collagen III in the CFs with or without Ang II stimulation,whereas BPTES significantly decreased the above indices in the CFs.α-KG supplementation reversed the inhibitory effect of BPTES on the CFs under Ang II stimulation.Furthermore,in vivo intraperitoneal injection of BPTES alleviated cardiac fibrosis of Ang II-treated mice.In conclusion,glutaminolysis plays an important role in the process of cardiac fibrosis induced by Ang II.Targeted inhibition of glutaminolysis may be a new strategy for the treatment of myocardial fibrosis.

关 键 词：谷氨酰胺代谢 谷氨酰胺酶1 心肌纤维化 

分 类 号：R542.23[医药卫生—心血管疾病]