Abstract：
The aging inflammatory microenvironment surrounding Leydig cells is linked to reduced testosterone levels in males. Tumor necrosis factor alpha-induced protein 3 (TNFAIP3) acts as a critical anti-inflammatory factor in various aging-related diseases. This study aims to investigate the protective effect of TNFAIP3 on testosterone production in Leydig cells under an aging inflammatory microenvironment.
Bioinformatics analysis examined TNFAIP3 expression differences in aging rat testes and validated the findings in aging mouse testes. In vitro models of inflammation were established using two Leydig cell lines, with tumor necrosis factor alpha (TNF-α) as the inflammatory factor. Lentiviral transduction was utilized to manipulate TNFAIP3 expression in these cell lines. Transcriptomic sequencing identified differentially expressed genes in TNFAIP3-overexpressing cells.
Bioinformatics analysis and validation experiments revealed increased inflammatory signaling and elevated TNFAIP3 expression in aging rat and mouse testes. TNFAIP3 knockdown worsened testosterone synthesis inhibition and apoptosis in cells, while TNFAIP3 overexpression reversed these effects. Transcriptome analysis identified alterations in the P38MAPK pathway following TNFAIP3 overexpression. TNFAIP3 knockdown enhanced TNF-induced P38MAPK signaling, whereas its overexpression attenuated this effect. TNFAIP3 was found to regulate testosterone synthesis by upregulating CEBPB expression.
TNFAIP3 exhibits inhibitory effects on apoptosis and promotes testosterone production in Leydig cells. The protective influence of TNFAIP3 on Leydig cells within an inflammatory microenvironment is likely mediated through by inhibiting the P38MAPK pathway and upregulating CEBPB expression.
Bioinformatics analysis examined TNFAIP3 expression differences in aging rat testes and validated the findings in aging mouse testes. In vitro models of inflammation were established using two Leydig cell lines, with tumor necrosis factor alpha (TNF-α) as the inflammatory factor. Lentiviral transduction was utilized to manipulate TNFAIP3 expression in these cell lines. Transcriptomic sequencing identified differentially expressed genes in TNFAIP3-overexpressing cells.
Bioinformatics analysis and validation experiments revealed increased inflammatory signaling and elevated TNFAIP3 expression in aging rat and mouse testes. TNFAIP3 knockdown worsened testosterone synthesis inhibition and apoptosis in cells, while TNFAIP3 overexpression reversed these effects. Transcriptome analysis identified alterations in the P38MAPK pathway following TNFAIP3 overexpression. TNFAIP3 knockdown enhanced TNF-induced P38MAPK signaling, whereas its overexpression attenuated this effect. TNFAIP3 was found to regulate testosterone synthesis by upregulating CEBPB expression.
TNFAIP3 exhibits inhibitory effects on apoptosis and promotes testosterone production in Leydig cells. The protective influence of TNFAIP3 on Leydig cells within an inflammatory microenvironment is likely mediated through by inhibiting the P38MAPK pathway and upregulating CEBPB expression.
摘要：
背景：睾丸间质细胞周围的老化炎症微环境与男性睾丸激素水平降低有关。肿瘤坏死因子α诱导蛋白3(TNFAIP3)是多种衰老相关疾病中的重要抗炎因子。本研究旨在探讨TNFAIP3对衰老炎症微环境下睾丸间质细胞睾酮产生的保护作用。
方法：生物信息学分析检查了衰老大鼠睾丸中TNFAIP3表达的差异，并验证了衰老小鼠睾丸中的发现。使用两种Leydig细胞系建立了炎症的体外模型，肿瘤坏死因子α(TNF-α)作为炎症因子。利用慢病毒转导来操纵这些细胞系中的TNFAIP3表达。转录组测序鉴定了TNFAIP3过表达细胞中差异表达的基因。
结果：生物信息学分析和验证实验显示，衰老大鼠和小鼠睾丸中炎症信号传导增加和TNFAIP3表达升高。TNFAIP3敲低恶化睾酮合成抑制和细胞凋亡，而TNFAIP3过表达逆转了这些效应。转录组分析鉴定了TNFAIP3过表达后P38MAPK途径的改变。TNFAIP3敲低增强TNF诱导的P38MAPK信号,而其过度表达减弱了这种作用。发现TNFAIP3通过上调CEBPB表达来调节睾酮合成。
结论：TNFAIP3对睾丸间质细胞的凋亡具有抑制作用，并促进睾丸激素的产生。TNFAIP3对炎症微环境中的Leydig细胞的保护性影响可能是通过抑制P38MAPK途径和上调CEBPB表达来介导的。
方法：生物信息学分析检查了衰老大鼠睾丸中TNFAIP3表达的差异，并验证了衰老小鼠睾丸中的发现。使用两种Leydig细胞系建立了炎症的体外模型，肿瘤坏死因子α(TNF-α)作为炎症因子。利用慢病毒转导来操纵这些细胞系中的TNFAIP3表达。转录组测序鉴定了TNFAIP3过表达细胞中差异表达的基因。
结果：生物信息学分析和验证实验显示，衰老大鼠和小鼠睾丸中炎症信号传导增加和TNFAIP3表达升高。TNFAIP3敲低恶化睾酮合成抑制和细胞凋亡，而TNFAIP3过表达逆转了这些效应。转录组分析鉴定了TNFAIP3过表达后P38MAPK途径的改变。TNFAIP3敲低增强TNF诱导的P38MAPK信号,而其过度表达减弱了这种作用。发现TNFAIP3通过上调CEBPB表达来调节睾酮合成。
结论：TNFAIP3对睾丸间质细胞的凋亡具有抑制作用，并促进睾丸激素的产生。TNFAIP3对炎症微环境中的Leydig细胞的保护性影响可能是通过抑制P38MAPK途径和上调CEBPB表达来介导的。
