Guerrero-Muñoz Jesús, Medina Lisvaneth, Castillo Christian, Liempi Ana, Fernández-Moya Alejandro, Araneda Sebastian, Ortega Yessica, Rojas-Pirela Maura, Maya Juan Diego, Kemmerling Ulrike
Programa de Biología Integrativa, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
Programa de Biología Integrativa, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile; Núcleo de Investigación Aplicada en Ciencias Veterinarias y Agronómicas, Facultad de Medicina Veterinaria y Agronomía, Universidad de Las Américas, Chile.
Placenta. 2023 Nov;143:117-123. doi: 10.1016/j.placenta.2023.10.009. Epub 2023 Oct 23.
Upon infection, Trypanosoma cruzi, a protozoan parasite, crosses the placental barrier and causes congenital Chagas disease. Ex vivo infection of human placental explants (HPEs) with the parasite induces apoptotic cell death. This cellular process involves changes in gene expression, which are partially regulated by miRNAs. In this study, we investigated the role of miR-512-3p, a highly expressed miRNA in the placenta, in parasite-induced apoptosis.
HPE cells were transfected with antagomirs or mimics of miR-512-3p and subsequently challenged with the parasite. The expression levels of miR-512-3p, caspase 3, caspase 8, and Livin were measured using RT-qPCR, and apoptotic cell death was analyzed based on caspase activity and DNA fragmentation assays.
Targeted inhibition of miR-512-3p effectively prevented parasite-induced expression and enzymatic activity of caspase 3 and caspase 8. However, it did not completely prevent DNA fragmentation, indicating the involvement of other factors in this process. Furthermore, the findings suggest that Livin may be regulated by miR-512-3p.
Our findings suggest that miR-512-3p modulates parasite-induced apoptosis in the trophoblast. By understanding the mechanisms involved in this process, we can gain insights into the pathogenesis of congenital Chagas disease and develop targeted therapeutic strategies.
感染后，原生动物寄生虫克氏锥虫会穿过胎盘屏障并导致先天性恰加斯病。用该寄生虫对人胎盘外植体（HPEs）进行体外感染会诱导细胞凋亡。这个细胞过程涉及基因表达的变化，这些变化部分受微小RNA（miRNAs）调控。在本研究中，我们调查了胎盘内高表达的miR - 512 - 3p在寄生虫诱导的细胞凋亡中的作用。
用miR - 512 - 3p的拮抗剂或模拟物转染HPE细胞，随后用寄生虫进行攻击。使用逆转录定量聚合酶链反应（RT - qPCR）测量miR - 512 - 3p、半胱天冬酶3、半胱天冬酶8和生存素的表达水平，并基于半胱天冬酶活性和DNA片段化分析来分析细胞凋亡。
对miR - 512 - 3p的靶向抑制有效阻止了寄生虫诱导的半胱天冬酶3和半胱天冬酶8的表达及酶活性。然而，它并未完全阻止DNA片段化，表明在此过程中还有其他因素参与。此外，研究结果表明生存素可能受miR - 512 - 3p调控。
我们的研究结果表明，miR - 512 - 3p调节滋养层中寄生虫诱导的细胞凋亡。通过了解这一过程中涉及的机制，我们可以深入了解先天性恰加斯病的发病机制并制定有针对性的治疗策略。
