Cytoskeletal protein SPTA1 mediating the decrease in erectile function induced by high-fat diet via Hippo signaling pathway
Cytoskeletal protein SPTA1 mediating the decrease in erectile function induced by high-fat diet via Hippo signaling pathway
复制标题
细胞骨架蛋白 SPTA1 通过 Hippo 信号通路介导高脂饮食引起的勃起功能下降
DOI：
10.1111/andr.13338
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发表时间：
2022-11-25
期刊：
影响因子：
4.5
通讯作者：
Chen，Jun
中科院分区：
文献类型：
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作者：
Chen，Ying;Wang，Lei;Chen，Jun
高脂饮食（HFD）诱导勃起功能下降的机制尚未被阐明，在先前的研究中，光谱蛋白α，红细胞1 （SPTA1）是一种调节细胞功能的细胞骨架蛋白，它属于一个通过调节Hippo信号通路上的效应物YAP来影响细胞和组织生长发育的蛋白家族，但其特殊作用尚未被阐明。目的探讨spta1在HFD致勃起功能异常中的作用。方法采用转录组学方法对正常饮食和高脂饮食小鼠阴茎组织进行转录组学分析，筛选差异表达基因，进一步鉴定大鼠阴茎组织中密切相关的靶基因，并在体外构建高糖（HG）处理的海绵体内皮细胞（CCECs）和海绵体平滑肌细胞（CCSMCs）模型中验证靶基因的表达。目的基因在阴茎组织不同细胞群中的分布通过单细胞测序男性健康图谱数据库检索。此外，通过干扰靶基因进一步探索勃起功能下降的机制。结果转录组学分析筛选出下调的差异基因espta1；Western blot和免疫组化结果显示，HFD组SD大鼠阴茎组织中SPTA1的表达明显降低。免疫荧光染色显示CD31和VWF在ccscs中阳性表达，α‐SMA在ccscs中阳性表达。SPTA1蛋白在CCECs和CCSMCs中HG组的表达水平显著降低。spta1mrna的表达在CCSMCs中显著降低，在cceccs中显著升高。spta1在不同的细胞群体中可能具有不同的表达模式和生物学功能。实时定量PCR结果显示，与对照siNC相比，转染CCSMCs的siSPTA1具有显著的干扰作用。将siSPTA1转染CCSMCs后，eNOS mRNA和蛋白表达显著下调，YAP、Caspase‐1、GSDMD、GSDMD‐N IL‐18和IL‐1β蛋白表达水平显著上调。CCSMCs收缩型蛋白α - SMA的表达水平显著下调。结论HFD喂养SD大鼠SPTA1下调可能通过激活Hippo通路诱导细胞热亡，导致勃起功能下降；这些发现可能为改善勃起功能提供新的治疗靶点。
BackgroundThe mechanism of high‐fat diet (HFD)‐induced decrease in erectile function has not been elucidated, and in previous studies, spectrin alpha, erythrocytic 1 (SPTA1) is a cytoskeletal protein that regulates cellular function, which belongs to a family of proteins that can affect cell and tissue growth and development by regulating YAP, an effector on the Hippo signaling pathway, but its particular role has not been elucidated.ObjectiveTo explore the role ofSPTA1in the abnormality of erectile function induced by HFD.MethodsWe analyzed the penile tissues of mice on normal diet and HFD by transcriptomics and screened for differentially expressed genes, further identified closely related target genes in rat penile tissues, and verified target gene expression in in vitro construction of high‐glucose (HG)‐treated corpus cavernosum endothelial cells (CCECs) and corpus cavernosum smooth muscle cells (CCSMCs) models. The distribution of target genes in various cell populations in penile tissues was retrieved by single‐cell sequencing Male Health Atlas database. Moreover, interfering with target genes was further applied to explore the mechanisms involved in erectile function decline.ResultsTranscriptomic analysis screened out down‐regulated differential geneSPTA1; Western blot and immunohistochemistry results showed that SPTA1 expression significantly decreased in the penile tissues of Sprague–Dawley (SD) rats in the HFD group. Immunofluorescence staining showed a positive expression of CD31 and VWF in CCECs and a positive expression of α‐SMA in CCSMCs. The expression level of SPTA1 protein significantly decreased in the HG group of CCECs and CCSMCs. The expression ofSPTA1mRNA significantly decreased in CCSMCs while significantly increased in CCECs.SPTA1may have various expression patterns and biological functions in different cell populations. Real‐time quantitative PCR results showed that the siSPTA1 transfected in CCSMCs had a significant interference effect compared with the control siNC. Transfection of siSPTA1 into CCSMCs resulted in the significant down‐regulation of mRNA and protein expression of eNOS, and significant up‐regulation of YAP, Caspase‐1, GSDMD, GSDMD‐N IL‐18, and IL‐1β protein expression levels. The expression level of CCSMCs contractile‐type protein α‐SMA was significantly down‐regulated.ConclusionsThe down‐regulation of SPTA1 in SD rats fed with HFD may induce cell pyroptosis and lead to the decrease of erectile function by activating the Hippo pathway; these findings may provide new therapeutic targets for improving erectile function.