基于Fas/FasL通路探讨参附注射液减轻细胞凋亡改善慢性心力衰竭  

Exploration on the effects of Shenfu Injection on apoptosis and improvement of chronic heart failure based on Fas/FasL pathway

作　　者：廉坤 李鑫 王菲 罗鹏 孟骊冲 李琳 胡志希 Lian Kun;Li Xin;Wang Fei;Luo Peng;Meng Lichong;Li Lin;Hu Zhixi(College of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,长沙410208

出　　处：《国际中医中药杂志》2025年第3期327-335,共9页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82274412、82305092);湖南省自然科学基金(2023JJ30453);湖南省中医药科研课题(B2023045);长沙市自然科学基金(kq2208185)。

摘　　要：目的基于Fas/FasL凋亡信号通路探讨参附注射液治疗慢性心力衰竭(CHF)的作用机制。方法将70只SPF级雄性C57BL/6小鼠按随机数字表法分为空白组15只和造模组55只。造模组采用腹腔注射异丙肾上腺素法制备CHF模型。4周后,将造模成功小鼠按随机数字表法分为模型组、参附组和西药组。相应药物干预15 d后,采用心脏彩超检测各组小鼠左室射血分数(LVEF)和左室缩短分数(LVFS);采用ELISA法检测小鼠血清氨基末端脑钠肽前体(NT-proBNP)水平;采用HE染色法观察小鼠心肌组织形态学变化;采用TUNEL pod法观察细胞凋亡情况,计算细胞凋亡率;采用RT-PCR法检测心肌组织TNF-α、Fas、Fas配体(FasL)、Caspase-3、Caspase-8 mRNA水平;采用Western blot法检测心肌组织TNF-α、Fas、FasL、Caspase-3、Caspase-8蛋白表达。将对数生长期的H9c2细胞按随机数字表法分为空白组、模型组和参附组。空白组细胞正常培养48 h;模型组细胞以80μmol/L异丙肾上腺素干预48 h;参附组细胞采用5μl/ml参附注射液预处理3 h后,加入80μmol/L异丙肾上腺素干预48 h。显微镜观察细胞生长情况;Western blot法及RT-PCR法检测相关蛋白表达与mRNA水平,TUNEL pod法和流式细胞术检测细胞凋亡情况。结果与模型组比较,参附组和西药组小鼠心功能改善,血清NT-proBNP水平降低(P<0.01或P<0.05),心肌细胞损伤和细胞凋亡减轻(P<0.01或P<0.05),心肌组织TNF-α、Fas、FasL、Caspase-3、Caspase-8 mRNA水平和蛋白表达降低(P<0.01或P<0.05)。空白组H9c2细胞呈长梭形,结构清楚;模型组心肌细胞皱缩变短,细胞边界模糊,呈现凋亡和坏死形态;参附组H9c2细胞多数呈梭形,细胞死亡减少,密度增大。与模型组比较,参附组H9c2细胞TNF-α、Fas、FasL、Caspase-3、Caspase-8蛋白及mRNA水平降低(P<0.01或P<0.05),细胞凋亡率降低(P<0.01)。结论参附注射液可能通过Fas/FasL信号通路改善CHF小鼠心功能,降低心肌细胞凋亡�Objective To explore the mechanism of Shenfu Injection in treating chronic heart failure(CHF)based on Fas/FasL apoptosis signaling pathway.Methods A total of 70 SPF male C57BL/6 mice were divided into blank group(15 mice)and model group(55 mice)according to random number table.The CHF model was prepared by intraperitoneal injection of isoproterenol.After 4 weeks,the successfully modeled mice were randomly divided intoa model group,Shenfu Injection group,and Western medicine group using a random number table method.After adfministration for 15 d,the left ventricular ejection fraction(LVEF)and left ventricular shortening fraction(LVFS)of each group of mice were measured by heart color ultrasound 1;serum NT-proBNP was detected by enzyme-linked immunosorbent assay(ELISA);Hematoxylin-eosin staining(HE)was used to observe the changes of myocardial tissue morphology;TUNEL pod method was used to detect apoptosis;the mRNA transcription levels of tumor necrosis factor-α(TNF-α),Fas,Fas ligand(FasL),Caspase-3 and Caspase-8 were detected by RT-PCR;protein expression levels of TNF-α,Fas,FasL,Caspase-3 and Caspase-8 in myocardial tissue were detected by Western blot.The logarithmically grown H9c2 cells were divided into blank group,model group and Shenfu Injection group.The cells in blank group were cultured for 48 hours;cells in the model group were exposed to 80μmol/L isoproterenol for 48 hours;cells in Shenfu Injection group were pretreated with 5μl/ml Shenfu injection for 3 hours and exposed to 80μmol/L isoproterenol for 48 h.The cell growth was observed under microscope.Western blot and RT-PCR were used to detect the expression of related proteins and mRNA transcription.TUNEL pod and flow cytometry were used to detect apoptosis.Results Compared with the model group,the cardiac function improved in the model group and Shenfu Injection group,serum NT ProBNP levels decreased(P<0.01 or P<0.05),myocardial cell injury and apoptosis were reduced(P<0.01 or P<0.05),and the mRNA and protein expression of TNF-α,Fas,FasL,Caspas

关 键 词：心力衰竭 参附注射液 Fas/FasL通路 细胞凋亡 小鼠 

分 类 号：R73[医药卫生—肿瘤]