摘要
目的:有关鹅掌楸苷对于缺血再灌注(ischemia/reperfusion,I/R)后心肌细胞是否具有保护作用及其作用机制的研究较少。本研究拟探讨鹅掌楸苷对I/R大鼠心功能的保护作用及其可能的机制。方法:将大鼠随机分为假手术组、I/R组、鹅掌楸苷+I/R组和丹参滴丸+I/R组。鹅掌楸苷+I/R组及丹参滴丸+I/R组于术前5 d至术后3 d,每日予以鹅掌楸苷溶液灌胃1次,I/R组及假手术组采用每日10 mL/kg生理盐水灌胃。术后分别对假手术组、I/R组和鹅掌楸苷+I/R组动物进行超敏肌钙蛋白定量检测。术后3 d应用超声心动图对大鼠心功能进行检测。处死大鼠后行苏木精-伊红(hematoxylin-eosin,HE)染色和免疫组织化学(immunohistochemistry,IHC)染色。进一步应用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)对组织行白介素(interleukin,IL)-1β、肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)、IL-6、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)、活性氧(reactive oxygen species,ROS)和乳酸脱氢酶A(lactate dehydrogenase A,LDHA)进行检测。应用蛋白质印迹法检测组织的炎症通路及凋亡相关蛋白的表达。4组大鼠均采用real-time PCR检测相关基因分子的相对mRNA表达。结果:与I/R组比较,术后2 h鹅掌楸苷+I/R组肌钙蛋白显著降低(P<0.05)。心脏超声显示:与I/R组比较,鹅掌楸苷+I/R组的心功能显著好转,病理学染色显示I/R组的心肌组织破坏严重,大量炎症细胞浸润。ELISA结果显示:与I/R组比较,鹅掌楸苷+I/R组的IL-1β、TNF-α、IL-6、MCP-1、ROS和LDHA含量均降低(均P<0.05)。与I/R组比较,鹅掌楸苷+I/R组炎症及凋亡相关蛋白表达均降低(均P<0.05)。与假手术组比较,I/R组、鹅掌楸苷+I/R组和丹参滴丸+I/R组B-淋巴细胞瘤-2基因(B-cell lymphoma-2,BCL-2)的mRNA相对表达量显著降低(均P<0.05);与I/R组比较,鹅掌楸苷+I/R组BCL-2 mRNA相对表达量显著升高(P<0.05)。与假手术组比较,I/R组BCL-2-相关X蛋白质(BCL-2 associated X protein,Bax)、半胱氨酸天冬氨酸蛋白酶(caspase,Cas)3、Cas9、κB抑制蛋白激酶(inhibitor ofκB kinase,IKK)α、IKKβ、核因子-κB(nuclear factor kappa B,NF-κB)、P65、P50、胞内磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)的mRNA相对表达量均显著升高(均P<0.05),鹅掌楸苷+I/R组和丹参滴丸+I/R组上述基因的mRNA相对表达量显著降低(均P<0.05);且鹅掌楸苷+I/R组显著低于I/R组(均P<0.05)。结论:鹅掌楸苷可能通过抑制局部炎症反应和细胞凋亡,进而对大鼠缺血再灌注后心功能起保护作用。
Objective:Research on whether liriodendrin exerts a protective effect on myocardial cells following ischemia/reperfusion(I/R)injury and its underlying mechanisms is limited.This study aims to explore the protective effects and potential mechanisms of liriodendrin on cardiac function in rats subjected to I/R injury.Methods:Rats were randomly divided into 4 groups:A sham surgery group,a I/R group,a liriodendrin+I/R group,and a Danshen dripping pill+I/R group.The liriodendrin+I/R and Danshen dripping pill+I/R groups received liriodendrin solution by gavage once daily from 5 days before surgery to 3 days after surgery,whereas the I/R group and the sham group received 10 mL/kg saline daily by gavage.Postoperatively,high-sensitivity cardiac troponin levels were quantified in the sham surgery group,the I/R group,and the liriodendrin+I/R group,and echocardiography was performed on day 3 to assess cardiac function.After euthanasia,hematoxylin-eosin(HE)staining and immunohistochemistry(IHC)staining were performed.Enzyme-linked immunosorbent assay(ELISA)was used to measure interleukin(IL)-1 beta(IL-1β),tumor necrosis factor-alpha(TNF-α),IL-6,monocyte chemoattractant protein-1(MCP-1),reactive oxygen species(ROS),and lactate dehydrogenase A(LDHA)levels in cardiac tissues.Western blotting was used to evaluate the expression of inflammation and apoptosis-related proteins,while real-time PCR was employed to assess relative mRNA expression levels of target genes across all four groups.Results:Compared with the I/R group,cardiac troponin levels in the liriodendrin+I/R group were significantly reduced 2 hours postoperatively(P<0.05).Echocardiography showed significant cardiac function improvement in the liriodendrin+I/R group.Pathological staining revealed severe myocardial damage and extensive inflammatory cell infiltration in the I/R group.ELISA results showed that IL-1β,TNF-α,IL-6,MCP-1,ROS,and LDHA levels were significantly lower in the liriodendrin+I/R group compared to the I/R group(all P<0.05).Additionally,inflammation and apoptosis-related protein expression levels were also significantly reduced in the liriodendrin+I/R group(all P<0.05).Compared with the sham group,mRNA levels of B-cell lymphoma-2(BCL-2)were significantly lower in the I/R,liriodendrin+I/R group,and Danshen dripping pill+I/R groups(all P<0.05);however,BCL-2 mRNA expression was significantly higher in the liriodendrin+I/R group compared to the I/R group(P<0.05).Furthermore,compared with the sham group,the I/R group exhibited significantly elevated mRNA levels of BCL-2-associated X protein(Bax),caspase-3(Cas3),Cas9,IκB kinase(IKK)α,IKKβ,nuclear factor kappa B cells(NF-κB),P65,P50,phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt)(all P<0.05);whereas these levels were significantly reduced in the liriodendrin+I/R and Danshen dripping pill+I/R groups(all P<0.05),with the liriodendrin+I/R group showing significantly lower levels than that the I/R group(all P<0.05).Conclusion:Liriodendrin may exert cardioprotective effects against I/R injury in rats by inhibiting local inflammatory responses and apoptosis.
作者
姚博宸
秦名震
马晓中
陈庆良
李博
郭志刚
YAO Bochen;QIN Mingzhen;MA Xiaozhong;CHEN Qingliang;LI Bo;GUO Zhigang(Department of Cardiac Surgery,Tianjin Chest Hospital,Tianjin 300001,China)
出处
《临床与病理杂志》
2024年第11期1506-1515,共10页
Journal of Clinical and Pathological Research
关键词
冠心病
缺血再灌注
鹅掌楸苷
细胞凋亡
炎症因子
炎症通路
coronary heart disease
ischemia/reperfusion
liriodendrin
apoptosis
inflammatory cytokines
inflammatory pathways
