山东医药2023，Vol.63Issue(29)：45-50,6.DOI:10.3969/j.issn.1002-266X.2023.29.010
当归多糖改善大鼠心肌缺血再灌注损伤的体内外观察及其机制探讨
Angelica sinensis polysaccharide ameliorates MIRI in rats in vitro and its mechanism
摘要
Abstract
Objective To establish in vivo and in vitro models of myocardial ischemia-reperfusion injury(MIRI)in rats,to observe the improvement effect of Angelica polysaccharide(ASP)on MIRI rats and to explore its mechanism.Methods In vivo study,adult SD rats were randomly divided into the sham group,MIRI group,low-dose ASP group and high-dose ASP group.Except the sham group,the MIRI models were established by ligation and reperfusion of the left an-terior descending branch of coronary artery in the other groups.After modeling,the rats in the low-dose and high-dose ASP groups were given 50 and 100 mg/kg ASP by intragastric administration,once a day,for 4 weeks,respectively.Rats in the sham group and MIRI group were given the equal volume of normal saline.Serum markers of myocardial injury such as creatine kinase-MB(CK-MB),lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin6(IL-6),and IL-1β were detected by ELISA.The expression levels of apoptotic proteins Bax,Bcl-2,cleaved Caspase-3,and TLR4/NF-κB pathway-related proteins TLR4,p-p65,and p-IκBα were detected by Western blotting.In vitro study,H/R cardiomycetes(derived from neonatal cell culture)were divided into the control group,hypoxia reoxygenation(H/R)group,low-dose H/R+ASP group and high-dose H/R+ASP group.The control group was cultured normally without treat-ment,and in vitro cardiomyocyte H/R models were established in the other three groups.After modeling,the low-dose and high-dose H/R+ASP groups were added with 5 and 10 µg/mL ASP,respectively,and the H/R group was not treated.MTT colorimetric assay was used to detect cardiomyocyte activity,and ELISA was used to detect inflammatory cytokines TNF-α,IL-6 and IL-1β.Western blotting was used to detect apoptotic proteins Bax,Bcl-2,cleaved Caspase-3,and TLR4/NFκB pathway proteins TLR4,p-p65,and p-IκBα in each group.Results In vivo,the levels of serum myocardi-al injury markers(LDH and CK-MB)and serum inflammatory factors(TNF-α,IL-6 and IL-1β)ranked from high to low:MIRI group,low-dose ASP group,high-dose ASP group,and sham group(all P<0.05).Myocardial Bax and cleaved Cas-pase-3 protein expression ranked from high to low:MIRI group,low-dose ASP group,high-dose ASP group,sham group,and myocardial Bcl-2 protein expression ranked from high to low:sham group,high-dose ASP group,low-dose ASP group,MIRI group(all P<0.05).The expression levels of TLR4/NF-κB pathway-related proteins TLR4,P-P65 and P-iκBα in the myocardial tissues were higher in the MIRI group than in the low-dose ASP group,which were higher than those of the high-dose ASP group,followed by the sham operation group(all P<0.05).In vitro study,cardiomyocyte activity ranked from high to low:control group,high-dose H/R+ASP group,low-dose H/R+ASP group,and H/R group(all P<0.05);the levels of cytokines TNF-α,IL-6 and IL-1β in H/R group were higher than those in H/R+ASP low-dose group,which were higher than those in the high-dose H/R+ASP group,followed by the control group(all P<0.05).Bax and cleaved caspase-3 protein expression ranked from high to low:H/R group,low-dose H/R+ASP group,high-dose H/R+ASP group,and control group.Cardiac Bcl-2 protein expression ranked from high to low:control group,high-dose H/R+ASP group,low-dose H/R+ASP group,and H/R group(all P<0.05);the expression levels of TLR4/NF-κB pathway-related proteins(TLR4,P-P65 and P-iκbα in cardiomyoma cells)in the H/R group were higher than those in the low-dose H/R+ ASP,which were higher than those of the high-dose H/R+ASP group,and those were higher in the high-dose H/R+ASP group than the control group(all P<0.05).Conclusion ASP can improve MIRI in both in vivo and in vitro ischemia-re-perfusion models,and its mechanism may be related to inhibiting the activation of TLR4/NFκB pathway,alleviating in-flammation,and reducing apoptosis.关键词
当归多糖/TLR4/NF-κB信号通路/炎症反应/细胞凋亡/心肌缺血再灌注损伤Key words
Angelica sinensis polysaccharide/Toll-like receptor 4/nuclear factor κB/inflammation/apoptosis/myocardial ischemia-reperfusion injury分类
医药卫生引用本文复制引用
叶建华,沈盛晖,张田杰,冯频频..当归多糖改善大鼠心肌缺血再灌注损伤的体内外观察及其机制探讨[J].山东医药,2023,63(29):45-50,6.基金项目
浙江省中医药科技计划科研基金项目(2021ZA032). （2021ZA032）
