SENP-1在慢性间歇性低氧诱导大鼠心肌损伤中的作用  

The role of SENP-1 in chronic intermittent hypoxia induced myocardial injury in rats

作　　者：贾远航 谭小武 陈林 涂容芳 周芳 Jia Yuanhang;Tan Xiaowu;Chen Lin;Tu Rongfang;Zhou Fang(Dept of Respiratory and Critical Care Medicine,The Second Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001)

机构地区：[1]南华大学衡阳医学院附属第二医院呼吸与危重症医学科,衡阳421001

出　　处：《安徽医科大学学报》2023年第10期1666-1672,共7页Acta Universitatis Medicinalis Anhui

基　　金：湖南省卫生健康委科研课题基金(编号:202203024555)。

摘　　要：目的探讨小泛素样修饰(SUMO)特异性蛋白酶1(SENP-1)对慢性间歇性低氧(CIH)诱导大鼠心肌损伤的影响及机制。方法将32只雄性SD大鼠分为对照组、CIH组、阴性对照腺相关病毒干预(AAV-shNC)组和SENP-1 shRNA腺相关病毒干预(AAV-shSENP-1)组,每组8只。CIH诱导6周后,进行超声心动图检查;ELISA检测血清中肌钙蛋白I(cTNI)、肌酸激酶MB同工酶(CKMB)、肌红蛋白(Mb)、乳酸脱氢酶(LDH)和心肌组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、白细胞介素(IL)-1β、IL-6和肿瘤坏死因子α(TNF-α)水平;HE染色观察心肌组织病理变化;DCFH-DA荧光探针标记法检测心肌组织活性氧(ROS)水平;试剂盒法检测心肌组织中低氧诱导因子-1α(HIF-1α)蛋白SUMO化水平;qRT-PCR和Western blot法检测心肌组织SENP-1和HIF-1αmRNA及蛋白表达水平。结果与对照组比较,CIH组大鼠心肌组织病理损伤严重,左心室舒张阶段末期内径(LVEDD)、左心室收缩阶段末期内径(LVESD)及血清cTNI、CKMB、Mb和LDH水平均升高(P<0.05),心肌组织中ROS、MDA、IL-1β、IL-6、TNF-α水平及SENP-1和HIF-1αmRNA及蛋白表达水平升高(P<0.05),而左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)及血清GSH和SOD水平降低(P<0.05),心肌组织HIF-1α蛋白SUMO化水平降低(P<0.05)。与CIH组比较,AAV-shSENP-1组大鼠心肌组织病理损伤减轻,LVEDD、LVESD及血清cTNI、CKMB、Mb和LDH水平降低(P<0.05),心肌组织中ROS、MDA、IL-1β、IL-6、TNF-α水平及SENP-1和HIF-1αmRNA和蛋白表达水平降低(P<0.05),LVEF、LVFS和血清GSH、SOD水平升高(P<0.05),心肌组织HIF-1α蛋白SUMO化水平升高(P<0.05)。结论抑制SENP-1表达可减轻CIH诱导的大鼠心肌炎症和氧化应激水平,改善心肌损伤和心功能障碍,其作用机制可能与提高HIF-1αSUMO化水平,进而抑制HIF-1α表达有关。Objective To investigate the effects and mechanism of small ubiquitin-like modifier(SUMO)specific proteinase-1(SENP-1)on chronic intermittent hypoxia(CIH)induced myocardial injury in rats.Methods 32 male SD rats were randomly divided into:control group,CIH group,negative control adeno-associated virus intervention group(AAV-shNC)and SENP-1 shRNA adeno-associated virus intervention group(AAV-shSENP-1),with 8 rats in each group.After 6 weeks of CIH induction,echocardiography was performed.The levels of cardiac troponin I(cTNI),creatine kinase MB isoenzyme(CKMB),myoglobin(Mb),lactate dehydrogenase(LDH)in serum and malondialdehyde(MDA),uperoxide dismutases(SOD),glutathione(GSH),interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in myocardial tissue were detected by ELISA.The pathological changes of myocardial tissue was observed by HE staining.The reactive oxygen species(ROS)level in myocardial tissue was detected by DCFH-DA fluorescence probe labeling.The small ubiquitin-like modifier(SUMO)level of hypoxia inducible factor-1α(HIF-1α)protein in myocardial tissue was detected by kit.The mRNA and protein levels of SENP-1 and HIF-1αin myocardial tissue were detected by qRT-PCR and Western blot.Results Compared with the control group,the pathological damage of myocardial tissue in CIH group was serious,the levels of left ventricular end diastolic diameter(LVEDD),left ventricular end systolic dimension(LVESD)and serum cTNI,CKMB,Mb and LDH significantly increased(P<0.05),and the levels of ROS,MDA,IL-1β,IL-6,TNF-αand the mRNA and protein levels of SENP-1 and HIF-1αin myocardial tissue also significantly increased(P<0.05),while the levels of LVEF,LVFS,serum GSH and SOD significantly decreased(P<0.05),and the SUMOylates level of HIF-1αprotein in myocardial tissue also significantly decreased(P<0.05).Compared with CIH group,AAV-shSENP-1 group had less myocardial pathological damage,the levels of LVEDD,LVESD and serum cTNI,CKMB,Mb and LDH significantly decreased(P<0.05),and the levels of ROS,MDA,IL-1β,IL-6,TNF

关 键 词：慢性间歇性低氧 小泛素样修饰特异性蛋白酶1 心肌损伤 低氧诱导因子-1Α 

分 类 号：R766[医药卫生—耳鼻咽喉科]