[1]赵明慧,薛 凌.毛蕊异黄酮靶向高迁移率族蛋白1对心肌缺血再灌注损伤大鼠保护作用实验研究[J].陕西医学杂志,2024,(7):873-878,883.[doi:DOI:10.3969/j.issn.1000-7377.2024.07.002]
　ZHAO Minghui,XUE Ling.Protective effect of calycosin targeting high mobility group box 1 on myocardial ischemia-reperfusion injury in rats[J].,2024,(7):873-878,883.[doi:DOI:10.3969/j.issn.1000-7377.2024.07.002]
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毛蕊异黄酮靶向高迁移率族蛋白1对心肌缺血再灌注损伤大鼠保护作用实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年7期
页码:
873-878,883
栏目:
基础研究
出版日期:
2024-07-05

文章信息/Info

Title:
Protective effect of calycosin targeting high mobility group box 1 on myocardial ischemia-reperfusion injury in rats
作者:
赵明慧; 薛 凌
(锦州医科大学附属第三医院心内科,辽宁 锦州 121001)
Author(s):
ZHAO Minghui; XUE Ling
(Department of Cardiology,Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China)
关键词:
心肌缺血再灌注损伤;  毛蕊异黄酮;  高迁移率族蛋白1;  炎症;  细胞凋亡;  Toll样受体4/核因子-κB通路
Keywords:
Myocardial ischemia-reperfusion injury;  Calycosin;  High mobility group box-1 protein;  Inflammation;  Apoptosis;  TLR4/NF-κB pathway
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2024.07.002
文献标志码:
A
摘要:
目的:探讨毛蕊异黄酮对心肌缺血再灌注损伤(MIRI)大鼠的保护作用及机制。方法:SPF级SD大鼠,建立心肌缺血再灌注损伤大鼠模型。随机分为四组:假手术组、心肌缺血再灌注损伤组、毛蕊异黄酮低剂量组、毛蕊异黄酮高剂量组。将H9C2细胞分为四组:对照组、缺氧/复氧组、毛蕊异黄酮组、重组高迁移率族蛋白1(HMGB1)蛋白组。超声心动图评估大鼠心功能,ELISA法测定心肌标志酶,HE染色观察心肌组织病理变化,TUNEL染色测定心肌组织和H9C2细胞凋亡情况,PPI网络分析毛蕊异黄酮的靶点预测和分子对接,胆囊收缩素八肽(CCK)8检测细胞活力,免疫荧光检测心肌HMGB1的表达量,Western blot检测高迁移率族蛋白1(HMGB1)、Toll样受体4(TLR4)、核因子-κB p65(NF-κB p65)和半胱氨酸蛋白酶-3(Cleaved caspase-3)相对表达水平。结果:心肌缺血再灌注损伤组较假手术组左心室舒张阶段末期内径(LVEDD)、左心室收缩阶段末期内径(LVESD)均上调(均P<0.05),而左心室射血分数(LVEF)和左心室短轴缩短率(LVFS)均下调(均P<0.05); 心肌纤维断裂,炎性细胞浸润增多; 血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)和肌钙蛋白(cTnI)水平显著升高; TUNEL阳性细胞数量升高,Cleaved caspase-3水平上调; HMGB1表达量增加(均P<0.05)。与心肌缺血再灌注损伤组比较,毛蕊异黄酮低、高剂量组LVEDD、LVESD 均下调(均P<0.05),而 LVEF 和 LVFS 均上调(均P<0.05); 心肌组织纤维排列尚规则,炎性细胞数量较少; 血清 CK-MB、LDH和cTnI水平降低; TUNEL阳性细胞数量降低,Cleaved caspase-3水平下调; HMGB1表达量降低(均P<0.05)。PPI和分子对接显示毛蕊异黄酮与HMGB1能较好的结合。缺氧/复氧组较对照组H9C2 细胞TUNEL细胞阳性数量显著升高,Cleaved caspase-3、HMGB1、TLR4、NF-κB p65表达上调(均P<0.05)。毛蕊异黄酮组较缺氧/复氧组H9C2 细胞TUNEL细胞阳性数量降低,Cleaved caspase-3、HMGB1、TLR4、NF-κB p65表达下调(均P<0.05)。而毛蕊异黄酮组基础上给予rHMGB1后,毛蕊异黄酮的保护作用被逆转。结论:毛蕊异黄酮能够明显改善大鼠心肌缺血再灌注损伤,这可能与抑制HMGB1/TLR4/NF-κB通路有关。
Abstract:
Objective:To probe the phylactic impact and mechanism of calycosin on myocardial ischemia-reperfusion injury(MIRI)in rats.Methods:A rat model of myocardial ischemia-reperfusion injury was established in SPF SD rats.They were randomly divided into four groups:sham group,MIRI group,low-dose calycosin group and high-dose calycosin group.H9C2 cells were divided into four groups:control group,hypoxia/reoxygenation group,calycosin group and HMGB1 protein group.Cardiac function was evaluated by echocardiography.ELISA was used to measure myocardial marker enzymes.HE staining was used to observe the pathological changes of myocardial tissue.TUNEL staining was used to detect the apoptosis of myocardial tissue and H9C2 cells.PPI network was used to analyze the target prediction and molecular docking of calycosin.Cell viability was detected by CCK8 assay.The expression of HMGB1 in myocardium was detected by immunofluorescence.Western blot was used to detect the relative expression levels of HMGB1,Toll-like receptor 4(TLR4),nuclear factor-κB p65(NF-κB p65)and Cleaved caspase-3.Results:Compared with the sham group,LVEDD and LVESD were up-regulated,while LVEF and LVFS were down-regulated in MIRI group(all P<0.05).Myocardial fibers were disrupted and inflammatory cell infiltration increased.Serum CK-MB,LDH and cTnI levels were significantly increased,the number of TUNEL positive cells and the level of Cleaved caspase-3 were increased,and the expression of HMGB1 was increased(all P<0.05).Compared with the MIRI group,LVEDD and LVESD in the low- and high-dose calycosin groups were down-regulated(all P<0.05),while LVEF and LVFS were up-regulated(all P<0.05),and the arrangement of myocardial fibers was still regular,and the number of inflammatory cells was small,and the serum levels of CK-MB,LDH and cTnI decreased,and the number of TUNEL positive cells and the level of Cleaved caspase-3 were decreased,and the expression of HMGB1 was decreased(all P<0.05).PPI and molecular docking showed that calycosin could bind to HMGB1.Compared with the control group,the number of TUNEL positive H9C2 cells in the hypoxia/reoxygenation group was significantly increased,and the expression of Cleaved caspase-3,HMGB1,TLR4 and NF-κB p65 was up-regulated(all P<0.05).Compared with the hypoxia/reoxygenation group,the number of TUNEL positive cells and the expression of Cleaved caspase-3,HMGB1,TLR4 and NF-κB p65 were decreased in the calycosin group(all P<0.05).However,the protective effect of calycosin was reversed when calycosin was added to rHMGB1 in the calycosin group.Conclusion:Calycosin can significantly improve MIRI in rats possibly by inhibiting the HMGB1/TLR4/NF-κB pathway.

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备注/Memo

备注/Memo:
基金项目:辽宁省自然科学基金资助项目(20170540372); 锦州医科大学附属第三医院青苗院长基金资助项目(FSSYQM2020004)
更新日期/Last Update: 2024-07-04