Study on the Protective Effect and Mechanism of Yiqi Jiedu Decoction on 12 Gy60 Coγ-Ray Induced Ferroptosis in Sertoli Cells
Objective To study whether 12 Gy60 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 Gy60 Coγ irradiation.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 Fe2+ 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,Fe2+,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,Fe2+,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 Gy60 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 mechanism of action may be related to the GPX4/ACSL4/LPCAT3 signaling pathway.
NETL
NSTL
万方数据
