TCONS_00016478 通过PGC1-α/ PPARγ信号通路影响实验性房颤兔心房肌能量代谢重构的机制

doi:10.6040/j.issn.1671-7554.0.2018.706

摘要/Abstract

摘要： 目的探讨长链非编码RNA(lncRNA)TCONS_00016478通过调控过氧化物酶增殖物激活受体γ 辅助激活因子1α(PGC-1α)/过氧化物酶增殖物激活受体γ(PPARγ)信号通路影响实验性房颤兔心房肌能量代谢重构的机制。方法采用高通量二代测序技术检测房颤兔/非房颤兔右心房组织差异性表达lncRNAs,成年新西兰白兔18只,体质量2.0~2.5 kg,雌雄不拘,随机分为假手术组(行开胸术但不注射病毒)、阴性对照慢病毒组(右心房注射阴性对照慢病毒)和TCONS_00016478沉默慢病毒组(右心房注射TCONS_00016478沉默慢病毒),每组6只。感染病毒前及感染1周后分别使用心脏电生理仪行程序电刺激,检测心房有效不应期(AERP)与房颤诱发性。感染病毒1周后处死动物,取心房肌组织,采用qRT-PCR法检测RNA的表达,采用Western blotting法检测蛋白质的表达,采用PAS染色法和油红O染色法分别检测糖原和脂滴沉积。结果与感染病毒前相比,感染病毒1周后,TCONS_00016478沉默慢病毒组AERP缩短(80.667±1.453 vs 71.750±2.411,t=3.168,P=0.034);假手术组(80.083±1.044 vs 79.333±0.333,t=0.684,P=0.531)与阴性对照慢病毒组(81.083±2.599 vs 80.000±2.646,t=0.022,P=0.983)手术前后AERP差异无统计学意义。TCONS_00016478沉默慢病毒组病毒感染1周后,3只诱发房颤,假手术组和阴性对照慢病毒组均未诱发房颤。假手术组、阴性对照慢病毒组及TCONS_00016478沉默慢病毒组心房肌TCONS_00016478(F=126.042,P<0.001)、PGC-1α(F=43.998,P<0.001)、PPARγ(F=417.863,P<0.001)、葡萄糖转运蛋白-4(GLUT4)(F=98.043,P<0.001)及碱棕榈酰转移酶-1(CPT1)(F=105.096,P< 0.001)基因表达量均差异有统计学意义。与假手术组相比,TCONS_00016478沉默慢病毒组心房肌TCONS_00016478在基因水平表达量降低(P<0.001),与能量代谢相关的蛋白质PGC-1α、PPARγ、GLUT4、CPT1在基因水平表达量降低(P<0.001),蛋白质水平表达量亦下降,差异有统计学意义(P<0.001);生物信息学分析表明,lncRNA TCONS_00016478及其靶基因PGC-1α 与心肌能量代谢密切相关。心房肌细胞糖原和脂滴异常沉积。结论 TCONS_00016478通过调控PGC-1α/PPARγ 信号通路影响心房肌能量代谢重构,进而调控房颤发生。

关键词: 心房颤动, 心房肌能量代谢重构, 长链非编码RNA, TCONS_00016478, 过氧化物酶增殖物激活受体γ 辅助激活因子1α, 过氧化物酶增殖物激活受体γ

Abstract: Objective To investigate the roles and mechanisms of TCONS_00016478 in atrial energy metabolic remodeling by regulating peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC-1α)/ peroxisome proliferator-activated receptor gamma(PPARγ)pathway in rabbit models of atrial fibrillation(AF). Methods The long non-coding RNA(lncRNA)expression profiles of right atria were investigated in AF and non-AF rabbit models using RNA sequencing technique. A total of 18 male and female adult New Zealand white rabbits weighting 2.0~2.5 kg were randomly divided into 3 groups: sham group(n=6, received thoracotomy only), negative control group(n=6, right atria infected with negative control lentiviruses), and lenti-RNAi-TCONS_00016478 group(n=6, right atria infected with TCONS_00016478 silencing lentiviruses). The atrial effective refractory period(AERP)and AF inducibility were measured before infection and 7 d after infection. After 7 d infection, the rabbits were sacrificed and atrial samples were collected. The gene and protein expressions were detected with qRT-PCR and Western blotting, respectively. The accumulation of glycogens and lipid droplets in atrial myocytes were assessed with periodic acid-Schiff(PAS)and Oil Red O staining, respectively. Results After 7 d infection, the AERP was significantly shortened in lenti-RNAi-TCONS_00016478 group(80.667±1.453 vs 71.750±2.411, t=3.168, P=0.034); however, no significant difference was found in the sham group(80.083±1.044 vs 79.333±0.333, t=0.684, P=0.531)and negative control group(81.083±2.599 vs 80.000±2.646, t=0.022, P=0.983). AF was induced in 3 rabbits in lenti-RNAi-TCONS_00016478 group, while no AF was induced in the sham and negative control groups. There were significant differences in the gene and protein expressions of TCONS_00016478(F=126.042, P<0.001), PGC1-α(F=43.998, P<0.001), PPARγ(F=417.863, P<0.001), glucose transporter type 4(GLUT4)(F=98.043, P<0.001) and carnitine palmitoyltransferase 1(CPT1)(F=105.096, P<0.001) among the 3 groups. Compared with the sham group, the lenti-RNAi-TCONS_00016478 group had lower expressions of TCONS_00016478, PGC1-α, PPARγ, GLUT4, and CPT1(all P<0.001), Bioinformatic analysis showed lncRNA TCONS_00016478 and PGC-1α were closely related to myocardial energy metabolism. But significantly increased accumulation of glycogens and lipid droplets in atrial myocytes. Conclusion TCONS_00016478 modulates atrial energy metabolic remodeling during AF through PGC-1α/PPARγ pathway.

Key words: Atrial fibrillation, Atrial energy metabolic remodeling, Long non-coding RNA, TCONS_00016478, Peroxisome proliferator-activated receptor gamma coactivator 1-alpha, Peroxisome proliferator-activated receptor gamma

中图分类号:

R541.7

李建华,李展,贾晓萌,杜娟娟,马神洲,刘东路,张勇,张玉娇,侯应龙. TCONS_00016478 通过PGC1-α/ PPARγ信号通路影响实验性房颤兔心房肌能量代谢重构的机制[J]. 山东大学学报 (医学版), 2019, 57(4): 1-8.

LI Jianhua, LI Zhan, JIA Xiaomeng, DU Juanjuan, MA Shenzhou, LIU Donglu, ZHANG Yong, ZHANG Yujiao, HOU Yinglong. Roles and mechanisms of TCONS_00016478 in atrial energy metabolic remodeling by regulating PGC-1α/ PPARγ pathway in rabbit models of atrial fibrillation[J]. Journal of Shandong University (Health Sciences), 2019, 57(4): 1-8.

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