金香丹抑制NLRP3/IL-1β/Caspase-1信号通路缓解大鼠心肌缺血再灌注损伤  

Jinxiangdan Inhibits NLRP3/IL-1β/Caspase-1 Pathway to Alleviate Myocardial Ischemia-reperfusion Injury in Rats

作　　者：萧闵[1] 向晶晶 王威[1] 王朝阳[1] XIAO Min;XIANG Jing-jing;WANG Wei;WANG Chao-yang(Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学,武汉430065

出　　处：《中国实验方剂学杂志》2021年第20期87-94,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖北省教育厅科学技术研究计划项目(B2017102)。

摘　　要：目的:观察金香丹预处理对心肌缺血再灌注损伤(MIRI)大鼠模型心肌组织NOD样受体热蛋白结构域3(NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)/白细胞介素-1β(IL-1β)信号通路的影响,探讨金香丹对MIRI的保护作用及其机制。方法:50只雄性SD大鼠随机分成假手术组,模型组,金香丹高、低剂量组及地奥心血康组,每组10只。造模前7 d开始,金香丹组给予金香丹片高、低剂量(0.72,0.18 g·kg^(-1))灌胃;假手术组和模型组每天给予等体积生理盐水灌胃;地奥心血康组给予地奥心血康(1.29 g·kg^(-1))灌胃。末次灌胃12 h后,结扎左冠状动脉前降支30 min,再灌注60 min的方法建立大鼠心肌缺血再灌注损伤模型。心电图检测ST段升高评价模型;比色法检测心脏组织肌酸激酶(CK)和乳酸脱氢酶(LDH)水平,苏木素-伊红(HE)染色观察心肌组织损伤情况,DNA断裂的原位末端标记(TUNEL)法检测心肌细胞凋亡情况,蛋白免疫印迹法(Western blot)检测心肌组织NLRP3,Caspase-1和IL-1β蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测心肌组织中NLRP3,Caspase-1和IL-1βmRNA的表达。结果:与假手术组比较,模型组心电图ST段显著升高(P<0.01),心肌组织CK和LDH活性显著增加(P<0.01),心肌组织凋亡细胞显著增加(P<0.01),NLRP3,IL-1β,Caspase-1蛋白和mRNA表达显著升高(P<0.01);与模型组比较,金香丹高、低剂量组及地奥心血康组心电图ST明显降低(P<0.05,P<0.01),心肌组织CK和LDH活性明显降低(P<0.05,P<0.01),心肌细胞凋亡明显改善(P<0.05,P<0.01),降低NLRP3,IL-1β,Caspase-1蛋白和mRNA表达水平(P<0.05,P<0.01);与地奥心血康组比较,金香丹低剂量组心电图ST段明显升高(P<0.05),金香丹高剂量组明显降低(P<0.05),金香丹高、低剂量组和地奥心血康组心肌组织绿色荧光强度明显降低(P<0.05,P<0.01),金香丹高剂量NLRP3,IL-1β,Caspase-1蛋白和mRNA表达均明显降低(P<0.05)。结论:金香丹可以降Objective:To observe the effect of Jinxiangdan(JXD)on NOD-like receptor pyrin domaincontaining-3(NLRP3)/cysteine-dependent aspartate-directed protease-1(Caspase-1)/interleukin-1β (IL-1β)signaling pathway in myocardium of rats with myocardial ischemia-reperfusion injury(MIRI)and explore the protective effect and mechanism of JXD against MIRI. Method:Fifty male SD rats were randomly divided into the sham operation group, model group, high-and low-dose JXD groups, and positive drug(Di’ao Xinxuekang)group,with 10 rats in each group. Seven days before modeling,rats in the JXD groups were separately treated with intragastric administration of 0.72 and 0.18 g·kg^(-1) JXD tablets,the ones in the sham operation group and model group with the same volume of normal saline,and those in the positive drug group with 1.29 g·kg^(-1) Di’ao Xinxuekang. Twelve hours after the last intragastric administration, the anterior descending branch of the left coronary artery was ligated for 30 min and then re-perfused for 60 min for inducing MIRI. ST segment elevation was detected by electrocardiogram(ECG)for model evaluation. The contents of creatine kinase(CK) and lactate dehydrogenase(LDH) in cardiac tissue were measured by colorimetry.Hematoxylin-eosin(HE)staining was conducted for observing myocardial histopathological changes,followed by the detection of cardiomyocyte apoptosis by DNA in situ nick end-labeling(TUNEL)assay. The protein and mRNA expression levels of NLRP3, Caspase-1, and IL-1β were detected by Western blot and real-time polymerase chain reaction(Real-time PCR),respectively. Result:Compared with sham operation group,the model group exhibited obviously elevated ST segment(P<0.01),enhanced CK and LDH activities in the myocardium(P<0.01),increased apoptotic cardiomyocytes(P<0.01),and up-regulated NLRP3,Caspase-1,and IL-1β protein and mRNA expression(P<0.01). Compared with model group,JXD at both the high and low doses and Di’ao Xinxuekang significantly lowered the ST segment(P<0.05,P<0.01),diminished the CK and

关 键 词：心肌缺血再灌注损伤 金香丹 NOD样受体热蛋白结构域3(NLRP3) 半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1) 细胞凋亡 

分 类 号：R2-0[医药卫生—中医学] R33