加味涤痰汤对慢性间歇低氧大鼠心肌铁死亡的影响  

Effect of Jiawei Ditan Decoction on cardiomyocyte ferroptosis in rats with chronic intermittent hypoxia

作　　者：王奕锦 陈沁 吴润华 林凡[1] 陈丽丽 伍娟 王春桔 杨眉峰 WANG Yijin;CHEN Qin;WU Runhua;LIN Fan;CHEN Lili;WU Juan;WANG Chunju;YANG Meifeng(College of Integrated Traditional Chinese and Western Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Clinical Skills Teaching Center of Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中西医结合学院,福州350122 [2]福建中医药大学临床技能教学中心

出　　处：《北京中医药大学学报》2024年第10期1427-1437,共11页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(No.82074192)。

摘　　要：目的基于磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路探讨加味涤痰汤对慢性间歇低氧大鼠心肌铁死亡的影响及其机制。方法将30只8周龄雄性SD大鼠以随机数字表法分为正常对照组、低氧干预组、加味涤痰汤干预组,每组10只。低氧干预组和加味涤痰汤干预组采用间歇低氧箱造模,每天干预8 h。每日干预前后予以灌胃,加味涤痰汤干预组药物剂量为16.38 g/kg,其余组予生理盐水,持续12周。HE染色观察心肌组织形态学;透射电镜观察心肌细胞线粒体超微结构;生化法检测心肌组织Fe^(2+)、丙二醛(MDA)含量;蛋白质印迹法检测心肌组织磷酸化PI3K(p-PI3K)、PI3K、磷酸化Akt(p-Akt)、Akt、核因子E2相关因子2(Nrf2)、谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)、酰基辅酶A合成酶长链家族成员4(ACSL4)蛋白表达。DMEM高糖培养基培养大鼠心肌细胞(H9c2);超高效液相色谱检测加味涤痰汤药物成分;CCK-8检测不同质量浓度加味涤痰汤干预下的细胞存活率。将细胞分为常氧组、铁死亡诱导剂组、铁死亡诱导剂+加味涤痰汤干预组,采用蛋白质印迹法检测p-Akt、GPX4蛋白表达;常氧组、低氧干预12 h组、低氧干预12 h+740Y-P组检测p-PI3K、GPX4蛋白表达;常氧组、低氧干预6 h组、低氧干预12 h组、低氧干预12 h+加味涤痰汤低、中、高剂量组(药物质量浓度分别为5、10和20 g/L),检测pPI3K、PI3K、p-Akt、Akt、GPX4蛋白表达。结果与正常对照组比较,低氧干预组大鼠心肌细胞排列紊乱、肿胀,线粒体排列混乱、膜密度增高、嵴断裂,同时伴有空泡变性;与低氧干预组比较,加味涤痰汤干预组心肌细胞排列较为整齐,未见明显肿胀,线粒体形态较为规则。与正常对照组比较,低氧干预组MDA、Fe^(2+)含量升高,p-PI3K/PI3K、p-Akt/Akt值降低,SLC7A11、GPX4、Nrf2表达减少,ACSL4表达升高(P<0.05);与低氧干预组比较,加味涤痰汤干预组MObjective To investigate the effect and mechanism of Jiawei Ditan Decoction on cardiomyocyte ferroptosis in rats with chronic intermittent hypoxia based on the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway.Methods Thirty 8-week-old male Sprague-Dawley rats were randomly divided into normal control,low-oxygen intervention,and Jiawei Ditan Decoction intervention groups using the random number table method,with 10 rats per group.The low-oxygen and Jiawei Ditan Decoction intervention groups were treated with intermittent hypoxia chamber for modeling,with 8 h of intervention daily.Gavage administration was performed before and after daily intervention.The intervention group received a dosage of 16.38 g/kg Jiawei Ditan Decoction,whereas the other two groups received normal saline.The experimental intervention period was 12 weeks.Hematoxylin and eosin staining was used to observe the morphology of myocardial tissue.Transmission electron microscope was used to observe the mitochondrial ultrastructure in cardiomyocytes.Biochemical detection was used to measure the Fe^(2+)and malondialdehyde(MDA)content in myocardial tissue.Phosphorylatedphosphoinositide 3-kinase(p-PI3K),PI3K,phosphorylated-protein kinase B(p-Akt),Akt,solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nrf2),and acyl-coenzyme A synthetase long-chain family member 4(ACSL4)protein expression in myocardial tissue were measured using western blotting.The rat cardiomyocyte cell line,H9c2,was cultured in high glucose Dulbecco′s Modified Eagle Medium,and the ultra-high-performance liquid chromatography was used to detect drug components in Jiawei Ditan Decoction.The CCK-8 assay was used to detect the cell survival rate of cells treated with different concentrations of Jiawei Ditan Decoction.The cells were divided into the normoxic,erastin,and erastin+Jiawei Ditan Decoction intervention groups,and p-Akt and GPX4 protein expression was measured using western blotting.pPI3K

关 键 词：加味涤痰汤 磷脂酰肌醇3激酶/蛋白激酶B 慢性间歇低氧 心肌 铁死亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]