摘要
目的探讨三七总皂苷通过调控NETs干预动脉粥样硬化伴急性心肌损伤的作用。方法以ApoE基因敲除(ApoE^(-/-))小鼠为研究对象,将21只8周龄ApoE^(-/-)小鼠随机分为ApoE^(-/-)组、ISO组、三七总皂苷(Panax Notoginseng Saponins,PNS组)给药组,每组7只。3组均饲喂高脂饲料模拟动脉粥样硬化(Atherosclerosis,AS)发病过程,给药组给予PNS组200 mg·kg^(-1)·d^(-1),每只100μL腹腔注射,ApoE^(-/-)组、ISO组每只腹腔注射0.9%生理盐水100μL,每天定点注射1次。8周后,ISO组和PNS组叠加腹腔注射异丙肾上腺素ISO(溶于0.9%生理盐水)10 mg·kg^(-1)·d^(-1),每只100μL,模拟急性心肌损伤。ISO连续造模并继续治疗5天后,处死取材。通过HE染色、油红O染色分别观察主动脉根部斑块病理形态学改变、心肌纤维化情况及脂质沉积情况;天狼星红染色评估心肌间隙胶原沉积情况;采用免疫组织化学法观察主动脉根部斑块部位中性粒细胞(Lymphocyte antigen 6 complex locus G6D,Ly6G)和髓过氧化酶(Myeloperoxidase,MPO)以及心肌中Ly6G、瓜氨酸化组蛋白(Citrullinated histone H3,Cit-H3)的表达情况。结果主动脉HE染色、油红O染色和天狼星红染色结果显示,与ApoE^(-/-)组相比,ISO组斑块面积与脂质沉积情况无显著差异(P>0.05),心肌纤维化面积和心肌间胶原纤维沉积面积显著增加(P<0.05);与ISO组相比,PNS组显著减小斑块面积、减少斑块上的脂质沉积、减少心肌纤维化面积和心肌间胶原纤维沉积(P<0.05)。免疫组织化学法结果提示,与ApoE^(-/-)组相比,ISO组斑块中MPO、Ly6G的表达无显著差异(P>0.05),心肌组织中Ly6G、Cit-H3表达明显升高(P<0.05);与ISO组相比,PNS组可以减少斑块中MPO、Ly6G的表达,并且降低心肌组织中Ly6G、Cit-H3表达(P<0.05)。结论PNS组对动脉粥样硬化伴急性心肌损伤具有显著抑制斑块和减轻心肌损伤的综合作用,其机制可能与抑制斑块与心肌损伤部位NETs的激活有关。
Objective To investigate the effect of total saponins of Panax ginseng in intervening atherosclerosis with acute myocardial injury through regulating NETs.Methods Twenty-one 8-week-old ApoE^(-/-)mice were randomly divided into ApoE^(-/-)group,ISO group and Panax Notoginseng Saponins(PNS)administration group,7 mice in each group.After 8 weeks,the ISO and PNS groups were superimposed with intraperitoneal injections of isoprenaline in ISO(dissolved in 0.9%saline).(dissolved in 0.9%saline)10 mg·kg^(-1)·d^(-1),100μL each,to simulate acute myocardial injury to ISO.After continuous moulding and continued treatment for 5 days,the samples were removed by execution.Pathomorphological changes,myocardial fibrosis and lipid deposition in the aortic root plaque were observed by HE staining and oil red O staining,respectively;collagen deposition in the myocardial space was assessed by Sirius red staining;immunohistochemistry was used to observe neutrophils in the aortic root plaque site(Ly6G)and myeloperoxidase(MPO),as well as the expression of Ly6G and citrullinated histone H3(Cit-H3)in the myocardium.Results The results of aortic H&E staining,oil red O staining and Sirius red staining showed no significant difference in plaque area and lipid deposition in the ISO group compared with the ApoE^(-/-)group(P>0.05),and a significant increase in myocardial fibrosis area and intermyocardial collagen fiber deposition area(P<0.05);compared with the ISO group,the PNS group significantly reduced plaque area,decreased lipid deposition on plaque Compared with the ISO group,the PNS group significantly reduced plaque area,reduced lipid deposition on plaque,reduced myocardial fibrosis area and interstitial collagen fiber deposition(P<0.05).The results of immunohistochemistry suggested that compared with the ApoE^(-/-)group,there was no significant difference in the expression of MPO and Ly6G in plaques in the ISO group(P>0.05),and the expression of Ly6G and Cit-H3 in myocardial tissue was significantly higher(P<0.05);compared with the ISO group,the PNS group could reduce the expression of MPO and Ly6G in plaques and decrease the expression of Ly6G and Cit-H3 in myocardial tissue.The PNS group reduced the expression of MPO and Ly6G in plaques and decreased the expression of Ly6G and Cit-H3 in myocardial tissue compared with the ISO group(P<0.05).Conclusion The PNS group had a combined effect of significantly inhibiting plaque and reducing myocardial injury in atherosclerosis with acute myocardial injury,and the mechanism may be related to the inhibition of the activation of NETs at the sites of plaque and myocardial injury.
作者
杨祯妮
李深广
陈瑜
熊敏琪
张腾
Yang Zhenni;Li Shenguang;Chen Yu;Xiong Minqi;Zhang Teng(Yueyang Hospital of Integrative Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Institute of Integrative Clinical Research,Shanghai Institute of Traditional Chinese Medicine,Shanghai 200437,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第5期1729-1735,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(82074049):人参皂苷Rb1抗心肌肥大的免疫相关-外泌体源性-非编码RNA调控机制研究,负责人:张腾
国家自然科学基金委员会青年科学基金项目(81903835):三七总皂苷对动脉粥样硬化伴急性心肌缺血的综合干预效应与miRNA调控机制研究,负责人:熊敏琪
