摘要
目的 研究叉头框蛋白O3(FoxO3)在心脏成纤维细胞(CF)衰老中的作用及机制。方法 选取新生Wistar乳鼠第五代CF随机分为小干扰RNA转染对照(siNC)组和siFoxO3组(n=3)。siFoxO3组转染FoxO3小干扰RNA。通过实时定量PCR和免疫印迹法检测衰老和Nod样受体蛋白3(NLRP3)炎性小体相关基因以及FoxO3表达情况。结果 与原代CF比较,第五代CF中细胞周期蛋白依赖激酶抑制剂2A(p16)和细胞周期蛋白依赖激酶抑制剂1A(p21)mRNA表达、p21和平滑肌肌动蛋白α(α-SMA)蛋白表达明显升高(P<0.05)。第五代CF中FoxO3 mRNA和蛋白表达明显低于原代(0.67±0.10 vs 1.03±0.02,0.47±0.12 vs 0.95±0.05,P<0.05)。与siNC组比较,siFoxO3组细胞中FoxO3 mRNA表达明显降低,而p16和p21 mRNA表达、p21蛋白表达明显上调(P<0.05,P<0.01)。与siNC组比较,siFoxO3组细胞中NLRP3、硫氧还蛋白结合蛋白、半胱氨酸天冬氨酸蛋白酶-1、白细胞介素1βmRNA表达明显升高(P<0.05,P<0.01)。结论 FoxO3通过调控NLRP3炎性小体抑制CF衰老,为衰老相关心血管疾病提供靶点。
Objective To investigate the role and mechanism of FoxO3 in the senescence of cardiac fibroblasts(CF).Methods The fifth passage(P5) CF of neonatal Wistar rats were randomly divided into control small interfering RNA transfection(siNC) group and siFoxO3 group(n=3).The cells of the siFoxO3 group were transfected with FoxO3 small interfering RNA.The expression of senescence and NOD-like receptor protein 3(NLRP3) inflammasome related genes and FoxO3 were detected by real-time PCR and Western blotting.Results Compared with the primary group, the mRNA expression of cyclin-dependent kinase inhibitor 2 A(p16) and cyclin-dependent kinase inhibitor 1 A(p21),and the protein expression of p21 and α-SMA were significantly increased in the P5 CF(P<0.05).The expression of FoxO3 at mRNA and protein levels was obviously decreased in P5 CF than in primary group(0.67±0.10 vs 1.03±0.02,0.47±0.12 vs 0.95±0.05,P<0.05).Compared with the siNC group, the mRNA level of FoxO3 was notably reduced, while the mRNA levels of p16,p21 and p21 protein level were remarkably up-regulated in the siFoxO3 group(P<0.05,P<0.01).Compared with the siNC group, the mRNA levels of NLRP3,thioredoxin binding protein, Caspase-1 and IL-1β were significantly increased in the siFoxO3 group(P<0.05,P<0.01).Conclusion FoxO3 inhibits CF senescence by regulating NLRP3 inflammasome, which may provide a target for aging-related cardiovascular diseases.
作者
师莹
张艺扉
赵莉莉
李曦铭
任珉
秦勤
Shi Ying;Zhang Yifei;Zhao Lili;Li Ximing;Ren Min;Qin Qin(Tianjin Institute of Cardiovascular Disease,Tianjin Chest Hospital,Tianjin 300222,China)
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2022年第12期1306-1309,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
天津市科技计划项目(20JCQNJC01980)
天津市医学重点学科(专科)建设项目(TJYXZDXK-055B)。
