上睑下垂
刺
免疫印迹
细胞生物学
流式细胞术
基因沉默
化学
分子生物学
程序性细胞死亡
细胞凋亡
生物
生物化学
基因
工程类
航空航天工程
作者
Q-F Deng,Ye Liu,Hong Wei Chu,Bou‐Yue Peng,Xiaofang Liu,Y.X. Cao
出处
期刊:Physiological Research
[Institute of Physiology of the Czech Academy of Sciences]
日期:2024-03-06
卷期号: (1/2024): 117-125
标识
DOI:10.33549/physiolres.935163
摘要
To explore the mechanism whereby cGAS-STING pathway regulates the pyroptosis of cryptorchidism cells, with a view to finding a new strategy for clinically treating cryptorchidism-induced infertility. Spermatogonial GC-1 cells were heat stimulated to simulate the heat hurt microenvironment of cryptorchidism. The cell viability was assayed by CCK-8, and cellular DNA damage was detected by γ-H2AX immunofluo-rescence assay. Flow cytometry was employed to assess pyroptosis index, while western blot, ELISA and PCR were used to examine the expressions of pyroptosis-related proteins (Caspase-1, IL-1β, NLRP3) and cGAS-STING pathway proteins (cGAS, STING). After STING silencing by siRNA, the expressions of pyroptosis-related proteins were determined. Pyroptosis occurred after heat stimulation of cells. Morphological detection found cell swelling and karyopyknosis. According to the γ-H2AX immunofluorescence (IFA) assay, the endonuclear green fluorescence was significantly enhanced, the γ-H2AX content markedly increased, and the endonuclear DNA was damaged. Flow cytometry revealed a significant increase in pyroptosis index. Western blot and PCR assays showed that the expressions of intracellular pyrogenic proteins like Caspase-1, NLRP3 and GSDMD were elevated. The increased STING protein and gene expressions in cGAS-STING pathway suggested that the pathway was intracellularly activated. Silencing STING protein in cGAS-STING pathway led to significantly inhibited pyroptosis. These results indicate that cGAS-STING pathway plays an important role in heat stress-induced pyroptosis of spermatogonial cells. After heat stimulation of spermatogonial GC-1 cells, pyroptosis was induced and cGAS-STING pathway was activated. This study can further enrich and improve the molecular mechanism of cryptorchidism.
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