摘要
目的研究参附注射液对自发性高血压大鼠(SHR)心肌损伤的影响并探讨E2相关因子2/血红素加氧酶1(Nrf2/HO-1)信号通路在其中的作用。方法SHR模型采用由遗传基因主导而自发形成的高血压大鼠,将40只SHR按随机数字表法分为五组,每组5只:模型组(腹腔注射生理盐水1/d)、卡托普利组(腹腔注射7 mg/kg卡托普利1/d,相当于人类的临床剂量)和参附注射液低、中、高剂量组(分别1/d腹腔注射1 ml/kg、2 ml/kg、4 ml/kg参附注射液,分别相当于人临床剂量的1/2、1、2倍)。另取8只WKY大鼠设为正常对照组(1/d腹腔注射生理盐水)。治疗4周后,通过动物无创血压计检测各组大鼠收缩压、舒张压和平均动脉压,超声检测各组大鼠心功能指标如左心室收缩期末期内径(LVESD)、左心室舒张期末期内径(LVEDD)、射血分数(EF)、短轴缩短率(FS)等,ELISA法检测各组大鼠血清心肌酶如天门冬氨酸氨基转移酶(AST)、肌酸激酶同工酶(CK-MB)、肌钙蛋白I(cTnI)水平,分光光度法检测各组血清氧化应激指标如丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)等,HE、TUNEL染色法检测各组大鼠心肌组织病变和细胞凋亡,透射电子显微镜观察各组心肌细胞超微结构变化,Western blot法检测各组心肌组织Nrf2/HO-1信号通路相关蛋白表达。结果与正常对照组比较,模型组大鼠收缩压、舒张压、平均动脉压、LVESD、LVEDD、AST、CK-MB、cTnI、MDA明显升高,EF、FS、SOD、CAT明显降低(P<0.05);模型组大鼠心肌纤维、心肌细胞呈现明显的病理学改变,凋亡细胞增多,凋亡指数(AI)明显升高(P<0.05);且模型组大鼠心肌组织Nrf2、HO-1、B淋巴细胞瘤-2(Bcl-2)蛋白表达量较正常对照组明显降低,Bcl-2相关X蛋白(Bax)、剪切型半胱氨酸蛋白酶-3(Cleaved Caspase-3)蛋白表达量及Bax/Bcl-2、Cleaved Caspase-3/Caspase-3比值较正常对照组明显升高(P<0.05)。与模型组比较,卡托普利组和参附注射液中剂量、高剂量组大鼠收缩压、舒张压、平均动脉压、LVESD、LVEDD、AST、CK-MB、cTnI、MDA明显降低(P<0.05),EF、FS、SOD、CAT明显升高(P<0.05)。且与模型组比较,卡托普利组和参附注射液低剂量、中剂量、高剂量组大鼠心肌组织病变和细胞凋亡状况不同程度改善,AI明显降低(P<0.05),心肌细胞超微结构改变不同程度改善。与模型组比较,卡托普利组和参附注射液中剂量、高剂量组大鼠心肌组织Nrf2、HO-1、Bcl-2蛋白表达量明显升高,Bax、Cleaved Caspase-3蛋白表达量及Bax/Bcl-2、Cleaved Caspase-3/Caspase-3比值明显降低(P<0.05)。参附注射液上述作用呈现一定的剂量依赖性。结论参附注射液具有减轻SHR心肌损伤、改善SHR心功能的作用,可能与激活Nrf2/HO-1信号通路,抑制氧化应激和细胞凋亡有关。
Objective To study the influence of Shenfu Injection(SFI)on myocardial injury in spontaneous hypertensive rats(SHR),and discuss the effect of signaling pathway of nuclear factor E2 related factor 2/heme oxygenase 1(Nrf2/HO-1).Methods SHR(n=40)were randomly divided into model group(given intraperitoneal injection of normal saline,once/d),captopril group(given intraperitoneal injection of captopril,7 mg/kg,once/d),and low-dose,mid-dose and high-dose SFI groups(given intraperitoneal injection of SFI,respectively in dose of 1 ml/kg,2 ml/kg,4 ml/kg,once/d,each n=5).And other 8 WKY rats were chosen into normal control group(given intraperitoneal injection of normal saline,once/d).After4-week treatment,the levels of systolic blood pressure(SBP),diastolic blood pressure(DBP)and mean arterial pressure(MAP)were detected in all groups.The indexes of heart function[left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),ejection fraction(EF),fraction shortening(FS)]were detected by using ultrasonic inspection.The indexes of serum myocardial enzyme[aspartate aminotransferase(AST),creatine kinase-MB isoenzyme(CK-MB)and cardiac troponin I(cTnI)]were detected by using ELISA.The indexes of serum oxidative stress[malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)]were detected by using spectrophotometry.The myocardial histopathological changes and cardiomyocyte apoptosis were detected by using HE or TUNEL staining.The ultrastructural changes of myocardial cells were observed by using transmission electron microscope(TEM).The expression of Nrf2/HO-1 signaling pathway related proteins were detected by using Western blotting assay.Results Compared with normal control group,SBP,DBP,MAP,LVESD,LVEDD,AST,CK-MB,cTnI,MDA increased significantly,and EF,FS,SOD and CAT decreased significantly in model group(P<0.05).The myocardial histopathological changes were observed in myocardial fibers and cardiomyocytes,cardiomyocyte apoptosis was higher and apoptosis index(AI)increased significantly in model group.The protein expressions of Nrf2,HO-1 and Bcl-2 decreased significantly,expressions of Bax and cleaved caspase-3 and ratio of Bax/Bcl-2 and cleaved caspase-3/caspase-3increased significantly in model group compared with normal control group(P<0.05).Compared with model group,SBP,DBP,MAP,LVESD,LVEDD,AST,CK-MB,cTnI,MDAdecreased significantly(P<0.05),and EF,FS,SOD and CAT increased significantly(P<0.05)in captopril groupand mid-dose and high-dose SFI groups.Compared with model group,myocardial histopathological changes and cardiomyocyte apoptosis were relieved in varying degrees and AI decreased significantly(P<0.05),and cardiomyocyteultrastructural changes were relieved in captopril groupand low-dose,mid-dose and high-dose SFI groups.Compared with model group,the protein expressions of Nrf2,HO-1 and Bcl-2 increased significantly,and expressions of Bax and cleaved caspase-3 and ratio of Bax/Bcl-2,and cleaved caspase-3/caspase-3 decreased significantly in captopril groupand mid-dose and high-dose SFI groups(P<0.05).The above effects of SFI showed a certain dose dependence.Conclusion SFI has the effects of relieving myocardial injury and improving heart function in SHR,which may be related to activation of Nrf2/HO-1 signaling pathway and inhibitions of oxidative stress and apoptosis.
作者
胡芳欣
孔凡英
张强
冯亮
李新峰
Hu Fangxin;Kong Fanying;Zhang Qiang;Feng Liang;Li Xinfeng(Department of Geriatrics,Handan Iron and Steel Hospital,Handan 056001,China;不详)
出处
《中国循证心血管医学杂志》
2025年第1期52-58,共7页
Chinese Journal of Evidence-Based Cardiovascular Medicine
基金
河北省医学科学研究课题计划(20211169)。
关键词
自发性高血压
参附注射液
心肌损伤
氧化应激
凋亡
Nrf2/HO-1通路
大鼠
Spontaneous hypertension
Shenfu Injection
Myocardial injury
Oxidative stress
Apoptosis
Nuclear factor E2 related factor 2/heme oxygenase 1 signaling pathway
Rats
作者简介
通讯作者:胡芳欣,E-mail:fengl0310@163.com。
