FOXO1调控PGC-1β/ERRα减轻LPS诱导的心肌细胞损伤  

FOXO1 regulates PGC-1β/ERRαto alleviate LPS-induced cardiomyocyte damage

作　　者：柯乐斌 金焕治 王蕾 KE Lebin;JIN Huanzhi;WANG Lei(Department of Occupational Health Examination,Wenzhou People's Hospital,Wenzhou,Zhejiang 325000,China;Department of General Practice,Wenzhou People's Hospital,Wenzhou,Zhejiang 325000,China)

机构地区：[1]浙江省温州市人民医院职业健康体检部,温州325000 [2]浙江省温州市人民医院全科医学,温州325000

出　　处：《浙江中西医结合杂志》2024年第12期1099-1105,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：温州市基础性科研项目(No.Y20220468)。

摘　　要：目的探讨叉头盒蛋白O1A(FOXO1)以及过氧化物酶体增殖物激活受体γ共激活因子-1β/雌激素相关受体α(PGC-1β/ERRα)通路在脓毒症诱导的心肌损伤中的作用机制。方法通过脂多糖(LPS)诱导H9C2细胞以模拟体外脓毒性心肌病模型。通过CCK-8检测细胞活性,流式细胞术检测细胞凋亡,酶联免疫吸附试验(ELISA)检测细胞上清液的炎症因子评估细胞损伤情况。利用JC-1探针检查测线粒体膜电位以评估线粒体损伤情况。蛋白免疫印迹实验和荧光定量PCR检测FOXO1、PGC-1β和ERRα的表达情况。结果与control组比较,LPS诱导的H9C2细胞活力[(39.26±4.32)%比(100.00±6.10)%,P<0.05]和FOXO1[(0.26±0.11)比(1.00±0.10),P<0.05]、PGC-1β[(0.18±0.03)比(1.00±0.08),P<0.05]、ERRα[(0.28±0.07)比(1.00±0.09),P<0.05]蛋白水平显著降低,凋亡水平[(17.32±0.49)%比(4.60±0.51)%,P<0.05]和炎症因子肿瘤坏死因子-α(TNF-α)[(268.38±11.14)pg/mL比(82.34±6.60)pg/mL,P<0.05]、白细胞介素-6(IL-6)[(143.65±8.41)pg/mL比(41.15±4.05)pg/mL,P<0.05]的释放显著升高。与LPS+oe-NC组比较,过表达FOXO1能促进PGC-1β[(3.98±0.16)比(0.99±0.19),P<0.05]、ERRα[(3.94±0.17)比(1.02±0.13),P<0.05]的表达和提高细胞活力[(294.64±13.71)%比(98.20±10.54)%,P<0.05],并减弱LPS诱导的线粒体损伤,抑制凋亡水平[(10.41±0.90)%比(16.68±1.24)%,P<0.05]和TNF-α[(129.90±10.13)pg/mL比(269.43±14.52)pg/mL,P<0.05]、IL-6[(69.55±7.35)pg/mL比(142.81±8.20)pg/mL,P<0.05]的释放水平。与LPS+oe-FOXO1组比较,应用ERRαantagonist-1后,PGC-1β[(1.03±0.20)比(3.98±0.16),P<0.05]和ERRα[(0.98±0.16)比(3.94±0.17),P<0.05]的蛋白表达以及细胞活力[(185.98±12.03)%比(294.64±13.71)%,P<0.05]下降,凋亡水平[(14.35±0.50)%比(10.41±0.90)%,P<0.05]和TNF-α[(197.74±8.00)pg/mL比(129.90±10.13)pg/mL,P<0.05]、IL-6[(105.75±7.27)pg/mL比(69.55±7.35)pg/mL,P<0.05]的释放水平上升。联合应用鱼藤酮/抗霉素A发现,与LPS+oe-FOXO1组比较,细胞活力[(100.64±12.0Objective To investigate the mechanism of forkhead box O1(FOXO1)and proliferator-activated receptor gamma co-activator 1β/estrogen-related receptorα(PGC-1β/ERRα)pathway in sepsis-induced myocardial injury.Methods H9C2 cells were induced by lipopolysaccharide(LPS)to simulate an in vitro model of sepsis-induced myocarditis.The cell viability was assessed using CCK-8 assay,the apoptosis was detected using flow cytometry, and the levels of inflammatory factors in the cell supernatant were measured using enzyme-linked immunosorbent assay to evaluate cell damage. Mitochondrial membrane potential was measured using JC-1 probe to evaluate mitochondrial damage. The protein expression levels of FOXO1, PGC-1β, and ERRα were detected using western blotting and fluorescence quantitative PCR. Results Compared with the control group, the LPS-induced H9C2 cells showed significantly reduced cell viability [(39.26±4.32)% vs. (100.00±6.10)%, P<0.05], along with decreased protein levels of FOXO1 [(0.26±0.11) vs. (1.00±0.10), P<0.05], PGC-1β [(0.18±0.03) vs. (1.00±0.08), P<0.05] and ERRα [(0.28±0.07) vs. (1.00±0.09), P<0.05]. Additionally, there was a significant increase in apoptosis levels [(17.32±0.49)% vs. (4.60±0.51)%, P<0.05] and the release of inflammatory factor tumor necrosis factor-α (TNF-α) [(268.38±11.14) vs. (82.34±6.60) pg/mL, P<0.05] and interleukin-6 (IL-6) [(143.65±8.41) vs. (41.15±4.05) pg/mL, P<0.05]. Compared with LPS+oe-NC group, FOXO1 overexpression increased the expression of PGC-1β [(3.98±0.16) vs. (0.99±0.19), P<0.05] and ERRα [(3.94±0.17) vs. (1.02±0.13), P<0.05], enhanced cell viability [(294.64±13.71)% vs. (98.20±10.54)%, P<0.05], attenuated LPS-induced mitochondrial damage, and inhibited apoptosis levels [(10.41±0.90)% vs. (16.68±1.24)%, P<0.05], and the release levels of TNF-α [(129.90±10.13) vs. (269.43±14.52) pg/mL, P<0.05] and IL-6 [(69.55±7.35) vs. (142.81±8.20) pg/mL, P<0.05]. Compared with LPS+oe-FOXO1 group, the application of ERRα antagonist-1 significantly

关 键 词：脓毒症 心肌损伤 FOXO1 PGC-1β/ERRα 线粒体 

分 类 号：R459.7[医药卫生—急诊医学] R542.2[医药卫生—治疗学]