定心方Ⅰ号方通过调控GSK3β/Nrf2/HO-1通路防治大鼠心肌缺血再灌注损伤的机制研究  

Dingxin RecipeⅠPrevents Myocardial Ischemia Reperfusion Injury through GSK3β/Nrf2/HO-1 Pathway

作　　者：刘晓瑜[1] 顾民华 徐煜凌 何培坤 洪睦铿 陈育尧[1] 贾钰华[1] LIU Xiaoyu;GU Minhua;XU Yuling;HE Peikun;HONG Mukeng;CHEN Yuyao;JIA Yuhua(College of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515 Guangdong,China)

机构地区：[1]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2020年第11期1265-1270,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省科技计划项目(2016A020226002);国家自然科学基金项目(81373574);广州市科技计划产学研协同创新重大专项项目(201704020042);广东省名优中成药二次开发项目(20174010)。

摘　　要：目的探究定心方Ⅰ号方(Dingxin RecipeⅠ,DXRⅠ)防治大鼠心肌缺血再灌注损伤(myocardial ischemia reperfusion injury,MIRI)的机制。方法将36只大鼠随机分为假手术组,MIRI模型组,胺碘酮组,定心方Ⅰ号方低、中、高剂量组。假手术组、MIRI模型组及胺碘酮组给予生理盐水灌胃,定心方Ⅰ号方各组大鼠分别给予不同剂量的定心方Ⅰ号方灌胃,连续灌胃7 d。在末次给药2 h后,采取结扎左前降支30 min后再复灌1 h的方法复制大鼠缺血再灌注模型。胺碘酮组大鼠术前10 min尾静脉注射5 mg·kg-1盐酸胺碘酮注射液。复制模型成功后取材,HE染色检测心脏组织病理形态学变化,采用相关试剂盒检测血清肌酸激酶同工酶(Creatine kinase isoenzymes,CK-MB)、乳酸脱氢酶(Lactate dehydrogenase,LDH)、大鼠心肌肌钙蛋白Ⅰ(Rat Cardiac TroponinⅠ,cTn-Ⅰ)、还原型谷胱甘肽(Reduced glutathione,GSH)、丙二醛(Malondialdehyde,MDA)和超氧化物歧化酶(Superoxide dismutase,SOD),Western Blot法检测心脏组织中糖原合成酶激酶-3(synthase kinase-3β,GSK3β)、磷酸化糖原合成酶激酶-3(Phosphorylated synthase kinase-3β,p-GSK3β)、转录因子NF-E2相关因子(Nuclear factor erythroid2-related factor 2,Nrf-2)、血红素氧合酶(Heme oxygenase-1,HO-1)、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2-associated X Protein,Bax)等蛋白的表达水平。结果HE染色结果显示,与假手术组比较,MIRI模型组心肌细胞出现了水肿和坏死,伴有局部出血和中性粒细胞浸润,而不同浓度的定心方Ⅰ号方可改善上述病理变化;相关检测试剂盒测定结果显示,与假手术组比较,MIRI模型组中的CK-MB、LDH、cTn-Ⅰ和MDA浓度明显升高(P<0.01),GSH和SOD活性明显下降(P<0.01),而定心方Ⅰ号方中、高剂量组可不同程度地抑制CK-MB、LDH、cTn-Ⅰ和MDA的表达(P<0.05,P<0.01),升高GSH和SOD活性(P<0.05,P<0.01);Western Blot结果显示,与MIRI模型组比较,不同浓度的�Objective To investigate the mechanism of Dingxin recipe Ⅰ(DXR Ⅰ) in preventing and treating myocardial ischemia-reperfusion injury(MIRI) of rats.Methods Thirty-six rats were randomly divided into Sham operation group,ischemia-reperfusion injury model group,Amiodarone(AM) group,DXR Ⅰ low,medium and high dose groups.Rats in each group of DXRI were given different doses of DXRI for 7 days,Sham operation group and ischemia-reperfusion injury model group were given normal saline,and rats in AM group were injected with 5 mg·kg-1 amiodarone hydrochloride in the tail vein before a surgical operation.Rat model of myocardial ischemiareperfusion injury was established after 30 min of reperfusion for 1 h.After copying the model successfully,the serum and tissue samples were taken.HE staining was used to detect pathological changes of cardiac tissue.Relevant commercial kits were used to detect CK-MB,LDH,cTn-Ⅰ,GSH,MDA,SOD in serum.Western Blot was used to detect the expression levels of GSK3β,p-GSK3β,Nrf-2,HO-1,Bcl-2 and Bax in heart tissue.Results HE staining showed that compared with Sham group,cerebral cells in the MIRI group showed edema and necrosis with local hemorrhage and neutrophil infiltration,while different doses of DXRI can improve the above pathological changes.The test results of related test kits showed that,compared with the Sham group,the concentrations of CKMB,LDH,cTn-Ⅰ and MDA in the MIRI group rats were significantly increased(P <0.01),and the activities of GSH and SOD were significantly decreased(P <0.01),while DXRI medium and high dose groups inhibited the expression of CK-MB,LDH,cTn-Ⅰ and MDA(P <0.05,P <0.01) and increased the activities of GSH and SOD(P <0.05,P <0.01).The results of Western Blot showed that compared with rats in the MIRI model group,different doses of DXRI could inhibit the expression of p-GSK3 and Bax protein(P <0.05,P <0.01),medium and high dose groups promote the expression of Nrf-2,HO-1 and Bcl-2 proteins(P <0.05,P <0.01).Conclusion DXRI can alleviate myocardial isc

关 键 词：定心方Ⅰ号方 心肌缺血再灌注损伤 氧化应激 GSK3β/Nrf2/HO-1信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]