hsa_circ_0091894/FLNA轴在β-氨基丙腈致小鼠主动脉损伤中的作用及机制研究  

Effect and mechanism of hsa_circ_0091894/FLNA axis in β-aminopropionitrile induced aortic injury in mice

作　　者：薛伍洋[1] 师天元[1] 田丰[1] XUE Wu-yang;SHI Tian-yuan;TIAN Feng(The Second Affiliated Hospital of Xinxiang Medical University,Xinxiang Henan 453002,China)

机构地区：[1]新乡医学院第二附属医院,河南新乡453002

出　　处：《毒理学杂志》2023年第1期37-42,共6页Journal of Toxicology

摘　　要：目的 探究hsa_circ_0091894/细丝蛋白A(filamin A, FLNA)轴在β-氨基丙腈(β-aminopropionitrile, BAPN)致小鼠主动脉损伤中的作用及机制。方法 将小鼠随机分为对照组、BAPN组、pcDNA3.1组和pcDNA3.1-hsa_circ_0091894组,对小鼠饲喂BAPN,尾静脉注射hsa_circ_0091894过表达的慢病毒载体。采用苏木素-伊红(HE)染色检测小鼠主动脉组织形态,采用实时荧光定量PCR(qRT-PCR)检测主动脉组织hsa_circ_0091894、肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)和白细胞介素-6(IL-6)表达水平,采用蛋白免疫印迹(Western blot)检测主动脉组织FLNA、B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、裂解的天冬氨酸特异性半胱氨酸蛋白酶3(cleaved Caspase-3)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)和α-1抗胰蛋白酶(α1-AT)蛋白表达水平,采用免疫荧光染色检测主动脉组织FLNA蛋白表达,采用原位末端标记法(TUNEL)染色检测主动脉组织细胞凋亡水平。结果 对照组、BAPN组、pcDNA3.1组和pcDNA3.1-hsa_circ_0091894组主动脉组织hsa_circ_0091894水平依次为1.00±0.00、0.29±0.05、0.34±0.07和0.83±0.11,FLNA蛋白水平依次为0.71±0.12、0.27±0.05、0.23±0.04和0.52±0.09。与对照组比较,BAPN组小鼠主动脉组织TUNEL阳性率,Bax/Bcl-2比值,cleaved Caspase-3、MMP-2和MMP-9蛋白表达水平,TNF-α、IFN-γ和IL-6表达水平明显增加(F=138.073,F=294.376,F=56.247,F=80.415,F=71.782,F=57.438,F=84.94,F=51.028,P<0.05);hsa_circ_0091894表达水平,FLNA和α1-AT蛋白表达水平明显降低(F=231.2,F=68.605,F=69.386,P<0.05)。与pcDNA3.1组比较,pcDNA3.1-hsa_circ_0091894组小鼠主动脉组织TUNEL阳性率,Bax/Bcl-2比值,cleaved Caspase-3、MMP-2和MMP-9蛋白表达水平,TNF-α、IFN-γ和IL-6表达水平明显降低;hsa_circ_0091894表达水平、FLNA和α1-AT蛋白表达水平明显增加(P<0.05)。结论 hsa_circ_0091894可减轻主动脉损伤,其机制可能与hsa_circ_0091894促进FLNA表达、降低细胞凋亡、炎症Objective To investigate the effect and mechanism of hsa_circ_0091894/filamin A(FLNA) axis in β-aminopropionitrile(BAPN) induced aortic injury in mice. Methods Mice were randomly divided into control group, BAPN group, pcDNA3.1 group and pcDNA3.1-hsa_circ_0091894 group. Mice fed with β-aminopropionitrile were injected lentivirus vector overexpressed of hsa_circ_0091894 into caudal vein. Hematoxylin-eosin(HE) staining was used to detect the aortic tissue morphology of mice. Real-time quantitative reverse transcriptionPCR(qRT-PCR) was used to detect the expression levels of hsa_circ_0091894, tumor necrosis factor-α(TNF-α), interferon-γ(IFN-γ) and interleukin-6(IL-6) of aortic tissue. Western blot was used to detect the protein expression levels of FLNA, B lymphocytoma-2-associated X protein(Bax), B lymphocytoma-2(Bcl-2), lytic aspartic acid specific cysteine protease 3(cleaved Caspase-3), matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9) and α-1 antitrypsin(α1-AT) of aortic tissue. Immunofluorescence staining was used to detect the protein expression of FLNA of aortic tissue. In situ end labeling(TUNEL) staining was used to detect the cell apoptosis of aortic tissue. Results The hsa_circ_0091894 levels in aortic tissue of control group, BAPN group, pcDNA3.1 group, pcDNA3.1-hsa_circ_0091894 group were 1.00±0.00, 0.29±0.05, 0.34±0.07 and 0.83±0.11, respectively. The FLNA protein levels were 0.71±0.12, 0.27±0.05, 0.23±0.04 and 0.52±0.09, respectively. Compared with the control group, the TUNEL positive rate, Bax/Bcl-2 ratio, cleaved Caspase-3, MMP-2, MMP-9 protein expression levels, TNF-α, IFN-γ, IL-6 expression levels in BAPN group of aortic tissue were significantly increased(F=138.073, F=294.376, F=56.247, F=80.415, F=71.782,F=57.438,F=84.94,F=51.028,P<0.05), while hsa_circ_0091894 expression levels, FLNA and α1-AT protein expression levels were significantly decreased(F=231.2, F=68.605, F=69.386, P<0.05). Compared with pcDNA3.1 group, the TUNEL positive rate, Bax/Bcl-2 ratio,

关 键 词：hsa_circ_0091894 细丝蛋白A β-氨基丙腈 主动脉损伤 凋亡 基质金属蛋白酶 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]