Chávez-Güitrón L E, Morales-Montor J, Nava-Castro K E, Ramírez-Álvarez H, Moreno-Mendoza N A, Prado-Ochoa M G, Muñoz-Guzmán M A, Alba-Hurtado F
División Químico-Biológicas, Universidad Tecnológica de Tecámac, Km. 37.5 Carretera Federal México Pachuca, Col. Sierra Hermosa, 55740, Tecámac, MEX, Mexico.
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, CDMX, 04510, Mexico.
Exp Parasitol. 2019 Mar;198:1-6. doi: 10.1016/j.exppara.2019.01.001. Epub 2019 Jan 8.
The in vitro effect of progesterone in T. canis larvae on their enlargement and motility were evaluated, together to the possible presence of progesterone receptors (PRs). T. canis larvae were cultured in RPMI-1640 with different concentrations of progesterone (0, 20, 40, 80, 400 and 800 ng/mL). Enlargement and increases in motility were dependent on the concentration only from 0 to 80 ng/mL (p < 0.05). The mean percentage of PR + cells in newly obtained larvae as measured by flow cytometry was 8.16 ± 0.4. The number of PR + cells increased depending on concentration from 0 to 80 ng/mL (p < 0.001). Cells obtained from larvae stimulated at any of the studied hormone concentrations showed greater mean fluorescence intensity when compared to non-stimulated cells. Additionally, the expression and location of PR + cells were determined in the larvae. The sequence of an amplicon (420-bp) obtained by PCR from T. canis larvae showed 100% homology with a gene fragment that codes for the PR of the dog. PR + cells were immunolocated using confocal microscopy in the intestinal region of the larvae that had been recently obtained. The results of this study show that T. canis larvae can recognize and respond to the presence of progesterone through a molecule possibly able to bind it. Since we previously observed a similar response to prolactin, we suggest that both hormones could participate sequentially in the reactivation of T. canis larvae in pregnant bitches.
评估了孕酮对犬弓首蛔虫幼虫增大和运动能力的体外作用，以及孕酮受体（PRs）可能的存在情况。犬弓首蛔虫幼虫在含有不同浓度孕酮（0、20、40、80、400和800 ng/mL）的RPMI-1640培养基中培养。仅在0至80 ng/mL的浓度范围内，幼虫的增大和运动能力的增加依赖于浓度（p < 0.05）。通过流式细胞术测量，新获得的幼虫中PR + 细胞的平均百分比为8.16 ± 0.4。PR + 细胞的数量在0至80 ng/mL的浓度范围内随浓度增加而增加（p < 0.001）。与未刺激的细胞相比，在任何研究的激素浓度下刺激获得的幼虫细胞显示出更高的平均荧光强度。此外，还确定了幼虫中PR + 细胞的表达和定位。通过PCR从犬弓首蛔虫幼虫获得的一个扩增子（420 bp）的序列与编码犬PR的基因片段显示出100%的同源性。使用共聚焦显微镜在新获得的幼虫的肠道区域对PR + 细胞进行免疫定位。本研究结果表明，犬弓首蛔虫幼虫能够通过一种可能与之结合的分子识别并响应孕酮的存在。由于我们之前观察到对催乳素的类似反应，我们认为这两种激素可能依次参与怀孕母犬体内犬弓首蛔虫幼虫的重新激活。
