当归红芪超滤物含药血清对放射性H9C2心肌细胞铜死亡的干预效应  

Intervention effect of Radix Angelicae Sinensis and Radix Hedysari serum on cuproptosis of radioactive H9C2cardiomyocytes

作　　者：汪鸣 赵信科[2] 刘生发 钱燕 张滋龙 安彩莲 张永茂 WANG Ming;ZHAO Xinke;LIU Shengfa(Zhangye People's Hospital Affiliated to Hexi University,Zhangye 734000,China;Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]河西学院附属张掖人民医院,甘肃张掖734000 [2]甘肃中医药大学,甘肃兰州730000

出　　处：《四川中医》2025年第3期138-143,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：甘肃省高校教师创新基金项目(编号:2023B-165);甘肃省青年科技基金项目(编号:23JRRG0042);河西学院校长基金青年科研项目(编号:QN2022018)。

摘　　要：目的:观察当归红芪超滤物(RAS-RH)含药血清对放射性H9C2心肌细胞铜死亡的干预效应。方法:SPF级Wistar雄性大鼠予RAS-RH(100mg·kg^(-1)·d^(-1))灌胃给药制备含药血清。将H9C2细胞随机分为对照组(正常血清干预)、模型组(6Gy X线照射+正常血清干预)及观察组(6Gy X线照射+含药血清干预)。采用CCK-8比色法检测各组H9C2心肌细胞增殖情况;Seahorse细胞能量代谢分析技术检测各组细胞线粒体糖酵解及氧化磷酸化水平;ELISA法检测各组细胞线粒体三磷酸腺苷(ATP)、谷胱甘肽(GSH)及丙酮酸脱氢酶(PDH)表达情况;ICP-MS法测定各组细胞铜元素含量;RT-PCR技术检测各组细胞铜死亡关键因子铁硫蛋白1(FDX1)、二氢硫辛酸转乙酰基酶(DLAT)及硫辛酸合成酶(LIAS)mRNA的表达情况。结果:与对照组比较,模型组培养48h及72h后细胞增殖情况、基础耗氧率、ATP合成量、最大耗氧率、氧储备能力、ATP、GSH及PDH表达均明显降低(P<0.05),细胞铜元素含量及铜死亡关键因子FDX1、DLAT、LIAS mRNA表达均显著升高(P<0.05)。与模型组比较,观察组培养48h及72h后细胞增殖情况、基础耗氧率、ATP合成量、最大耗氧率、氧储备能力、ATP、GSH及PDH表达均明显增加(P<0.05),铜元素含量及铜死亡关键因子FDX1、DLAT及LIAS mRNA表达均明显降低(P<0.05)。结论:铜死亡可能是辐射诱导心肌细胞损伤的重要机制,当归红芪超滤物通过改善放射性心肌细胞线粒体功能,抑制细胞铜死亡,起到保护放射性心肌损伤的作用。Objective: To observe the effect of Radix Angelicae Sinensis and Radix Hedysari (RAS-RH) serum on cuproptosis of H9C2cardiomyocytes. Methods: SPF-grade Wistar male rats were administrated intragastric with RAS-RH (100mg?kg -1 ?d -1 ) to prepare drug-containing serum. H9C2cells were randomly divided into control group (normal serum intervention),model group (6Gy X-ray irradiation + normal serum intervention) and observation group (6Gy X-ray irradiation + drug-containing serum intervention). The proliferation of H9C2cardiomyocytes in each group was detected by CCK-8colorimetry. The levels of mitochondrial glycolysis and oxidative phosphorylation were measured by Seahorse cell energy metabolism analysis. The expression of Adenosine triphosphate (ATP),Glutataione (GSH) and Pyruvate Dehydrogenase (PDH) in mitochondria of each group was detected by ELISA. The content of copper element in each cell was determined by ICP-MS. The mRNA expressions of Ferredoxin 1(FDX1),Dihydrolipoyl Transacetylase (DLAT) and Lipoic Acid Synthetase (LIAS),a key factor of cuproptosis,were detected by RT-PCR. Results: Compared with the control group,cell proliferation,basal oxygen consumption rate,ATP synthesis amount,maximum oxygen consumption rate,oxygen reserve capacity,ATP,GSH and PDH expression were significantly decreased in the model group after 48h and 72h culture ( P <0.05). The content of copper and the mRNA expressions of the key factors of cuproptosis FDX1,DLAT and LIAS were significantly increased ( P <0.05). Compared with model group,cell proliferation,basal oxygen consumption rate,ATP synthesis amount,maximum oxygen consumption rate,oxygen reserve capacity,ATP,GSH and PDH expression were significantly increased in observation group after 48h and 72h culture ( P <0.05). The copper content and the mRNA expressions of the key factors of cuproptosis FDX1,DLAT and LIAS were significantly decreased ( P <0.05). Conclusion: Cuproptosis may be an important mechanism of radiation-induced cardiomyocyte injury. Radix Angelicae Sinensis and

关 键 词：当归红芪超滤物 放射性心脏病 铜死亡 线粒体 

分 类 号：R285[医药卫生—中药学]