西红花苷上调miR-346对心肌缺血再灌注大鼠心肌损伤的保护作用  

Protective effect of miR-346 up-regulated by crocin on myocardial ischemia reperfusion injury in rats

作　　者：李红霞[1] 康滢蓥 李瑶[2] 喻文立[1] Li Hongxia;Kang Yingying;Li Yao;Yu Wenli(Department of Anesthesiology,Shuixi Hospital,Tianjin First Central Hospital,Tianjin 300190,China;School of Medicine,Nankai University,Tianjin 300041,China)

机构地区：[1]天津市第一中心医院水西院区麻醉科,天津300190 [2]南开大学医学院,天津300041

出　　处：《国际生物医学工程杂志》2024年第4期335-341,共7页International Journal of Biomedical Engineering

基　　金：中国初级卫生保健基金会临床专项科研项目(YLGX-MZ-2022008);天津市第一中心医院青年创新人才培养项目。

摘　　要：目的探究西红花苷上调微小RNA-346(miR-346)对心肌缺血再灌注大鼠心肌损伤的保护作用。方法通过开胸结扎左前降支冠状动脉后再灌注构建心肌缺血再灌注大鼠模型,50只雄性SD大鼠随机分为5组,每组10只,分别为假手术组、模型组、西红花苷组、miR-346阴性对照(miR-NC)组、miR-346抑制剂组。假手术组大鼠开胸,但不结扎冠状动脉。miR-NC组、miR-346抑制剂组大鼠在结扎左前降支冠状动脉前48 h通过尾静脉注射miR-NC或miR-346抑制剂。8 mg西红花苷用100 ml生理盐水溶解稀释。西红花苷组、miR-NC组、miR-346抑制剂组大鼠按照80 mg/kg灌胃,假手术组、模型组大鼠灌胃5 ml/kg生理盐水。每天1次,连续给药15 d。酶联免疫吸附试验检测血清中肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)水平,实时荧光定量PCR(qRT-PCR)检测miR-346表达水平,HE染色法检测心肌组织的病理变化,TUNEL染色法检测心肌细胞凋亡率,Western Blot检测凋亡相关蛋白表达。结果与假手术组相比,模型组大鼠血清中CK-MB、LDH水平,病理评分,心肌细胞凋亡率及裂解的含半胱氨酸的天冬氨酸蛋白水解酶-3(cleaved Caspase-3)和B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)表达水平均升高(均P<0.05),心肌组织中miR-346表达水平和Bcl-2水平均降低(均P<0.05);与模型组相比,西红花苷组大鼠血清中CK-MB、LDH水平,病理评分,心肌细胞凋亡率及cleaved Caspase-3和Bax表达水平均降低(均P<0.05),心肌组织中miR-346表达水平和Bcl-2水平均升高(均P<0.05);与miR-NC组相比,miR-346抑制剂组大鼠血清中CK-MB、LDH水平,病理评分,心肌细胞凋亡率及cleaved Caspase-3和Bax表达水平均降低(均P<0.05),心肌组织中miR-346表达水平和Bcl-2水平均升高(均P<0.05)。结论西红花苷可通过上调miR-346减轻心肌缺血再灌注大鼠心肌组织损伤和心肌细胞凋亡。Objective To explore the protective effect of miR-346 up-regulated by crocin on myocardial ischemia reperfusion injury in rats.Methods The rat model of myocardial ischemia reperfusion was constructed by open-chest ligation of the left anterior descending coronary artery followed by reperfusion.Fifty male SD rats were randomly divided into the sham operation group,model group,crocin group,miR-346 negative control(miR-NC)group,and miR-346 inhibitor group with 10 rats in each group.Rats in the sham operation group were only received thoracotomy without ligation.Rats in the miR-NC group and miR-346 inhibitor group were injected with miR-NC or miR-346 inhibitor through the tail vein 48 h prior to ligation of the left anterior descending coronary artery ligation.The crocetin reagent for gastric lavage was prepared by dissolving 8 mg of crocetin in 100 ml of physiological saline.Rats in the crocetin group,miR-NC group,and miR-346 inhibitor group were gavaged with crocetin reagent at 80 mg/kg.Rats in the sham operation group and model group were gavaged with saline at 5 ml/kg.The crocetin reagent and saline were gavaged once a day for 15 days.Serum levels of creatine kinase-MB(CK-MB)and lactate dehydrogenase(LDH)were detected by enzyme-linked immunosorbent assay(ELISA).The miR-346 expression level was detected by real-time fluorescence quantitative PCR(qRT-PCR).The pathological changes in cardiac muscle tissues were detected by HE staining.Cardiomyocyte apoptosis rate was detected by TUNEL staining.The expression levels of apoptosis-related proteins were detected by Western Blot.Results Compared with the sham operation group,the serum levels of CK-MB and LDH,pathological scores,cardiomyocyte apoptosis rate and expression of cleaved cysteinyl aspartate specific proteinase-3(cleaved Caspase-3)and B-cell lymphoma-2(Bcl-2)-associated X protein(Bax)were increased in the model group(all P<0.05),and the miR-346 expression level and Bcl-2 levels in myocardial tissues were decreased(all P<0.05).Compared with the model group,serum

关 键 词：心肌缺血再灌注 西红花苷 微小RNA-346 肌酸激酶同工酶 乳酸脱氢酶 

分 类 号：R542.2[医药卫生—心血管疾病]