益气解毒方对12 Gy^(60)Coγ射线诱导的支持细胞铁死亡的防护作用及机制研究  

Study on the Protective Effect and Mechanism of Yiqi Jiedu Decoction on 12 Gy^(60)Coγ-Ray Induced Ferroptosis in Sertoli Cells

作　　者：庄鑫丽 姜荟茜 徐旻灏 叶一帆 徐文慧[1] 王安[1] 王磊[1] 王舒冉 赵章弟 张文彦 张淑静[1] 胡素敏[1] ZHUANG Xinli;JIANG Huiqian;XU Minhao;YE Yifan;XU Wenhui;WANG An;WANG Lei;WANG Shuran;ZHAO Zhangdi;ZHANG Wenyan;ZHANG Shujing;HU Sumin(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《中国中医基础医学杂志》2024年第4期627-632,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(12075035,11675027)。

摘　　要：目的研究12 Gy^(60)Coγ射线对睾丸支持细胞(sertoli cells,SCs)铁死亡的影响,并探究益气解毒方干预对支持细胞铁死亡的防护机制。方法支持细胞分为空白(NC)组,模型(IR)组、铁死亡激动剂(Erastin)组、铁死亡抑制剂(Lip-1)组、益气解毒方高剂量(YQJD-H)组、益气解毒方低剂量(YQJD-L)组,分别按照相应的条件培养,除NC组和Erastin组外,其余各组细胞均使用12 Gy^(60)Coγ射线进行一次性照射,建立支持细胞辐射损伤模型。照射后24 h,用细胞计数试剂(cell counting kit-8,CCK-8)检测细胞活性;DCFH-DA荧光探针检测细胞内活性氧(reactive oxygen species,ROS)水平;铁离子比色法检测细胞内Fe^(2+)水平;ELISA法检测细胞上清液高迁移率族蛋白B1(high mobility group protein box-1,HMGB1)水平;JC-1荧光探针检测细胞线粒体膜电位改变;实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction,RT-qPCR)检测细胞GPX4、ACSL4、LPCAT3 mRNA的表达。结果照射后24 h,与NC组比较,Erastin组和IR组支持细胞活性及线粒体膜电位均显著下降(P<0.01),ROS、Fe^(2+)、HMGB1水平均明显升高(P<0.01),GPX4 mRNA表达水平下降(P<0.05);IR组ACSL4、LPCAT3 mRNA表达水平升高(P<0.05)。与IR组比较,YQJD-H组、YQJD-L组、Lip-1组的细胞活性均显著升高(P<0.01),ROS、Fe^(2+)、HMGB1水平均明显降低(P<0.01);YQJD-H组GPX4 mRNA表达水平升高(P<0.05),ACSL4、LPCAT3 mRNA表达水平下降(P<0.05)。结论12 Gy^(60)Coγ射线诱导支持细胞发生铁死亡,而益气解毒方能减轻支持细胞铁死亡,起到辐射防护作用,其作用机制可能与GPX4/ACSL4/LPCAT3信号通路有关。Objective To study whether 12 Gy^(60)Coγ-ray induces ferroptosis of sertoli cells(SCs),and to investigate the mechanism of Yiqi Jiedu decoction against irradiation-induced ferroptosis of Sertoli cells.Methods Sertoli cells were divided into the negative control(NC)group,ionizing radiation(IR)group,ferroptosis activator(Erastin)group,ferroptosis inhibitor(Lip-1)group,Yiqi Jiedu decoction high dose(YQJD-H)group,Yiqi Jiedu decoction low dose(YQJD-L)group.Sertoli cells in each group were cultured according to the corresponding conditions.Except for NC group and Erastin group,all other groups received a one-time,single dose of 12 Gy^(60)Coγirradiation.24 h after irradiation,cell viability was detected by CCK8 method.The level of reactive oxygen species(ROS)was detected by DCFH-DA fluorescent probe.The level of Fe^(2+)was detected by intracellular iron colorimetric method.ELISA was used to detect the level of high mobility group protein box-1(HMGB1)in cell supernatant.JC-1 fluorescent probe was used to detect cellular mitochondrial membrane potential changes.The GPX4,ACSL4 and LPCAT3 gene expression were detected by real time quantitative polymerase chain reaction(RT-qPCR).Results 24 h after irradiation,compared with NC group,both Erastin and IR groups showed a significant decrease in sertoli cell viability and mitochondrial membrane potential(P<0.01),a significant increase in the levels of ROS,Fe^(2+),and HMGB1(P<0.01),and a decrease in the expression of GPX4 mRNA(P<0.05);yet the expression of ACSL4 and LPCAT3 mRNA in the IR group went up(P<0.05).Compared with the IR group,cell viability was significantly higher in the YQJD-H,YQJD-L,and Lip-1 groups(P<0.01),and ROS,Fe^(2+),and HMGB1 levels were significantly lower(P<0.01).GPX4 mRNA expression increased in the YQJD-H group(P<0.05),and ACSL4 and LPCAT3 mRNA expression decreased(P<0.05).Conclusion 12 Gy^(60)Coγ-ray induced ferroptosis in sertoli cells.Yiqi Jiedu decoction could alleviate ferroptosis in sertoli cells to reduce the radiation injury on sertoli cells.Its

关 键 词：益气解毒方 电离辐射 支持细胞 铁死亡 

分 类 号：R285.5[医药卫生—中药学]