收稿日期: 2024-08-21
网络出版日期: 2024-12-23
基金资助
国家自然科学基金青年科学基金资助项目(81901530)
Inositol⁃trisphosphate 3⁃kinase B causes impaired testosterone synthesis in senescent leydig cells by inhibiting mitophagy
Received date: 2024-08-21
Online published: 2024-12-23
目的 探究肌醇-三磷酸3-激酶B（ITPKB）对衰老睾丸间质细胞睾酮合成障碍的作用和机制。 方法 使用GO、GSEA富集分析和蛋白质相互作用网络对公共数据集进行综合分析，确定与衰老相关疾病中睾丸组织中相关通路和基因的表达。D-半乳糖腹腔注射构建衰老小鼠模型；培养小鼠睾丸间质细胞（TM3），将细胞分为4组，对照组（NC组）、衰老组（D-gal组）、抑制剂组（GNF362组）和D-gal + GNF362组。ELISA法检测小鼠血清睾酮、卵泡刺激素和黄体生成素含量。q-PCR检测IL-1α、IL-6、TNF-α、ITPKB mRNA水平；免疫荧光检测StAR、3β-HSD、ITPKB、LC3蛋白表达；Western blot检测P53、P21、StAR、ITPKB蛋白表达。透射电子显微镜观察细胞内结构。 结果 生物信息学分析提示ITPKB在衰老小鼠睾丸中高表达。衰老小鼠血清睾酮水平较年轻组小鼠降低。衰老组小鼠较年轻组小鼠睾丸中IL-1α、IL-6、TNF-α、ITPKB mRNA水平及P53、P21、ITPKB蛋白水平升高；StAR、3β-HSD、LC3蛋白质水平及线粒体自噬水平下降。与D-gal组的细胞相比，D-gal + GNF362组细胞上清液睾酮水平升高，线粒体自噬水平显著提升。 结论 ITPKB通过抑制线粒体自噬导致衰老睾丸间质细胞睾酮合成障碍。
关键词： 迟发性性腺功能减退症; 睾酮缺乏; 肌醇-三磷酸3-激酶B; 衰老; 睾酮替代疗法; 线粒体自噬
李梓光 , 韦存 , 赵磊 , 周其赵 . 肌醇-三磷酸3-激酶B通过抑制线粒体自噬导致衰老睾丸间质细胞睾酮合成障碍[J]. 实用医学杂志, 2024 , 40(24) : 3427 -3437 . DOI: 10.3969/j.issn.1006-5725.2024.24.001
Objective To explore the effect and mechanism of ITPKB on testosterone synthesis disorder of senescent Leydig cells. Methods A comprehensive analysis of public datasets was performed using GO， GSEA enrichment analysis and protein interaction network to determine the expression of related pathways and genes in testicular tissue in aging-related diseases. D-galactose intraperitoneal injection to construct an aging mouse model； Mouse Leydig cells （TM3） were cultured， and the cells were divided into four groups， control group （NC group）， senescence group （D-gal group）， inhibitor group （GNF362 group） and D-gal+GNF362 group. The serum levels of testosterone， follicle-stimulating hormone and luteinizing hormone in mice were detected by ELISA. q-PCR was used to detect the mRNA levels of IL-1α， IL-6， TNF-α and ITPKB. Immunofluorescence was used to detect the protein expressions of StAR， 3β-HSD， ITPKB and LC3. Western blot was used to detect the protein expressions of P53， P21， StAR and ITPKB. Transmission electron microscopy to observe intracellular structures. Results Bioinformatics analysis showed that ITPKB was highly expressed in the testes of aging mice. The serum testosterone level of aging mice was lower than that of young mice. Compared with the younger group， the mRNA levels of IL-1α， IL-6， TNF-α and ITPKB in the testes of mice in the aging group were increased， and the protein levels of P53， P21 and ITPKB were increased. The levels of StAR， 3β-HSD， LC3 protein and mitophagy decreased. Compared with the cells in the D-gal group， the testosterone level in the supernatant of the D-gal+GNF362 group was increased， and the level of mitophagy was significantly increased. Conclusion ITPKB causes impaired testosterone synthesis in senescent Leydig cells by inhibiting mitophagy.
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