摘要
目的观察环状RNA SORBS2(circSORBS2)对脓毒症小鼠心肌损伤的改善作用,并探讨其机制。方法48只SPF级C57BL/6J雄性小鼠按照随机数字表法分为Sham组、模型组、sh-NC组和sh-circSORBS2组,每组12只。模型组、sh-NC组和sh-circSORBS2组小鼠均采用盲肠结扎(CLP)法构建脓毒症心肌损伤小鼠模型。sh-NC组和sh-circSORBS2组小鼠在CLP前1周尾静脉注射sh-NC(阴性对照)或sh-circSORBS2(沉默circSORBS2表达)腺相关病毒载体颗粒;Sham组仅进行开腹、关腹手术,未结扎和穿刺盲肠,未进行尾静脉注射。小鼠建模完成后,观察并比较各组小鼠生存率、心肌组织病理、心功能指标[左心室收缩压(LVSP)、左心室压力最大上升速率(+dp/dt_(max))、左心室压力最大下降速率(-dp/dt_(max))、左心室舒张末期压(LVEDP)、肌酸激酶同工酶MB(CK-MB)、心肌肌钙蛋白I(cTnI)]、心肌组织炎症因子[肿瘤坏死因子(TNF)-α、白细胞介素6(IL-6)]、心肌组织焦亡相关因子[NOD样受体热蛋白结构域相关蛋白3(NLRP3)、半胱天冬酶1(Caspase-1)、IL-1β、成孔蛋白D(GSDMD)]以及心肌组织中circSORBS2、miR-21-3p、Sma/MAD相关蛋白7(SMAD7)mRNA相对表达量。通过starBase靶基因预测网站预测circSORBS2、miR-21-3p和SMAD7的靶向关系,采用荧光素酶报告基因实验进行验证。结果与Sham组相比,模型组小鼠生存率、LVSP、+dp/dt_(max)、miR-21-3p均降低,LVEDP、-dp/dt_(max)、cTnI、CK-MB、TNF-α、IL-6、NLRP3、Caspase-1、IL-1β、GSDMD、circSORBS2、SMAD7 mRNA均升高(P均<0.05),心肌组织出现坏死。与sh-NC组相比,sh-circSORBS2组小鼠生存率、LVSP、+dp/dt_(max)、miR-21-3p均升高,LVEDP、-dp/dt_(max)、cTnI、CK-MB、TNF-α、IL-6、NLRP3、Caspase-1、IL-1β、GSDMD、circSORBS2、SMAD7 mRNA均降低(P均<0.05),心肌损伤程度减轻。双荧光素酶报告基因实验结果显示,circSORBS2靶向调控miR-21-3p,而miR-21-3p靶向调控SMAD7。结论抑制circSORBS2表达可改善脓毒症小鼠的心肌损伤,其机制可能与circSORBS2通过吸附miR-21-3p解除对SMAD7的抑制有关。
Objective To observe the effect of circular RNA SORBS2(circSORBS2)on myocardial injury in septic mice and to explore its mechanism.Methods Forty-eight SPF C57BL/6J male mice were randomly divided into Sham group,model group,sh-NC group and sh-circSORBS2 group,with 12 mice in each group.The mice in the model group,sh-NC group and sh-circSORBS2 group were treated with cecal ligation(CLP)to construct the mouse models of sepsis-in⁃duced myocardial injury.Mice in the sh-NC group and sh-circSORBS2 group were intravenously injected with sh-NC(neg⁃ative control)or sh-circSORBS2(silencing circSORBS2 expression)adeno-associated virus vector particles one week be⁃fore CLP.In the Sham group,only laparotomy and abdominal closure were performed without ligation and puncture of the cecum,and no tail vein injection was performed.After the mouse models were completed,the survival rate,myocardial histopathology,cardiac function indexes[left ventricular systolic pressure(LVSP),_(max)imum rate of left ventricular pres⁃sure increase(+dp/dt_(max)),_(max)imum rate of left ventricular pressure decrease(-dp/dt_(max)),left ventricular end-diastolic pressure(LVEDP),creatine kinase isoenzyme MB(CK-MB),and cardiac troponin I(cTnI)],myocardial tissue inflam⁃matory factors[tumor necrosis factor(TNF)-αand interleukin-6(IL-6)],myocardial tissue pyroptosis-related factors[NOD-like receptor hot protein domain-related protein 3(NLRP3),caspase-1(Caspase-1),IL-1β,and Gasdermin D(GSDMD)],and the relative expression levels of circRBSSO2,miR-21-3 p,and Sma/MAD-related protein 7(SMAD7)mRNA in the myocardial tissues of mice in each group were observed and compared.The targeting relationship of circ⁃SORBS2,miR-21-3 p and SMAD7 was predicted by starBase target gene prediction website,and verified by luciferase re⁃porter gene experiment.Results Compared with the Sham group,the survival rate,LVSP,+dp/dt_(max),and miR-21-3p decreased,while LVEDP,-dp/dt_(max),cTnI,CK-MB,TNF-α,IL-6,NLRP3,Caspase-1,IL-1β,GSDMD,circSORBS2,and SMAD7 mRNA increased(all P<0.05),and myocardial necrosis occurred in the model group.Compared with the sh-NC group,the survival rate,LVSP,+dp/dt_(max),and miR-21-3p increased,while LVEDP,-dp/dt_(max),cTnI,CK-MB,TNF-α,IL-6,NLRP3,Caspase-1,IL-1β,GSDMD,circSORBS2,and SMAD7 mRNA decreased(all P<0.05),and the degree of myocardial injury was alleviated in the sh-circSORBS2 group.The results of the dual-luciferase reporter gene assay showed that circSORBS2 targetedly regulated miR-21-3p,and miR-21-3p regulated SMAD7.Conclusion Inhibition of circSORBS2 expression can alleviate myocardial injury in septic mice,and the mechanism may be related to circSORBS2 relieving the inhibition of SMAD7 by adsorbing miR-21-3p.
作者
袁媛
刘晓娴
龙苏
杨国辉
汤娜娜
宁睿
方昊
欧晶
吴曦
李子巍
YUAN Yuan;LIU Xiaoxian;LONG Su;YANG Guohui;TANG Nana;NING Rui;FANG Hao;OU Jing;WU Xi;LI Ziwei(Emergency Department,The Affiliated Hospital of Guizhou Medical University,Guiyang 550001,China;不详)
出处
《山东医药》
2026年第1期60-65,共6页
Shandong Medical Journal
基金
贵州省卫生健康委科学技术基金项目(gzwkj2025-217)
贵州医科大学国家自然科学基金培育项目(20NSP024)。
