Background

Spermatozoa interactions with fallopian tubes may influence fertilization. The purpose was to investigate cytokines, chemokines and growth factors expression from human fallopian tube epithelial cells (OE-E6/E7) exposed to spermatozoa.

Methods

Fresh semen samples were obtained from 10 healthy normozoospermic men. Sperms were prepared and co-cultured with OE-E6/E7. The cell line without spermatozoa was considered as the control group. Afterwards, Expression of 84 cytokines from OE-E6/E7 cell line in the presence and absence of spermatozoa were measured using PCR-array. Quantitative PCR was performed on seven genes to confirm the results of PCR-array analysis. Differentially expressed genes were subjected to www.geneontology.org and www.pantherdb.org to perform GO enrichment and panther pathway analysis. The concentration of IL-8, IL-10, IL-1B and BMP-4 in culture medium were analyzed by ELISA.

Results

Sperm interaction with the epithelial cells resulted in a significant increase in expression of TGF-β2, BMP-4, IL-10, IL-9, and CD40LG markers. Moreover, expression of IL-16, IL-17F, SPP-1, CXCL-13, MSTN, IL-1A, IL-1B, IL-8, BMP-7, CSF-2, CSF-3, VEGF-A, OSM, LTA, TNF, TNFRSF11B, TNFSF11, CCL-11, CCL-20, CCL-24, CCL-3, CCL-8, CX3CL1 and CXCL-9 were considerably reduced in presence of spermatozoa. Panther pathway analysis discovered 3 pathways for upregulated genes including gonadotropin-releasing hormone receptor, TGF-beta and interleukin signaling pathways. Furthermore, 9 pathways were detected for down-regulated genes. Inflammation signaling pathway which is mediated by chemokine and cytokine contains the most number of genes.

Conclusion

This study indicates that sperm modifies expression of cytokines, chemokines and growth factors from OE-E6/E7. Moreover, altered genes expression are toward higher survival chance of the spermatozoa.

中文翻译：

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输卵管上皮细胞通过调节细胞因子和趋化因子对精子的免疫反应

背景

精子与输卵管的相互作用可能会影响受精。目的是研究暴露于精子的人输卵管上皮细胞 (OE-E6/E7) 的细胞因子、趋化因子和生长因子的表达。

方法

新鲜精液样本取自 10 名健康的精子正常男性。制备精子并与 OE-E6/E7 共培养。没有精子的细胞系被认为是对照组。之后，使用 PCR 阵列测量了 OE-E6/E7 细胞系在存在和不存在精子的情况下 84 种细胞因子的表达。对七个基因进行定量 PCR 以确认 PCR 阵列分析的结果。对差异表达的基因进行 www.geneontology.org 和 www.pantherdb.org 进行 GO 富集和 panther 通路分析。通过ELISA分析培养基中IL -8、IL - 10、IL-1B和BMP-4的浓度。

结果

精子与上皮细胞的相互作用导致TGF-β2、BMP-4、IL-10、IL-9和CD40LG标志物的表达显着增加。此外，IL-16、IL-17F、SPP-1、CXCL-13、MSTN、IL-1A、IL-1B、IL-8、BMP-7、CSF-2、CSF-3、VEGF-A、 OSM、LTA、TNF、TNFRSF11B、TNFSF11、CCL-11、CCL-20、CCL-24、CCL-3、CCL-8、CX3CL1和CXCL-9在精子存在时显着减少。Panther 通路分析发现了 3 条上调基因的通路，包括促性腺激素释放激素受体、TGF-β 和白细胞介素信号通路。此外，检测到下调基因的 9 条途径。由趋化因子和细胞因子介导的炎症信号通路含有最多的基因。

结论

这项研究表明，精子会改变 OE-E6/E7 中细胞因子、趋化因子和生长因子的表达。此外，改变的基因表达有助于提高精子的存活率。