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1.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 302-8, 2015.
Artículo en Inglés | WPRIM | ID: wpr-637113

RESUMEN

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.

2.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 302-308, 2015.
Artículo en Inglés | WPRIM | ID: wpr-331069

RESUMEN

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.


Asunto(s)
Animales , Ratas , Autofagia , Secuencia de Bases , Cartilla de ADN , Hipoxia , Técnicas In Vitro , Pulmón , Lisosomas , Fisiología , Terapia por Inhalación de Oxígeno , Reacción en Cadena en Tiempo Real de la Polimerasa
3.
National Journal of Andrology ; (12): 949-952, 2013.
Artículo en Chino | WPRIM | ID: wpr-268024

RESUMEN

Mesenchymal stem cells (MSCs) are somatic stem cells capable of self-renewing and multipotent differentiation. They are also referred to as " seed cells" in tissue engineering. Recent years have witnessed great advances in the studies of the differentiative potential and paracrine function of MSCs, as well as in their clinical applications in several fields. As for their application in male infertility, researches are still in the stage of animal experiments. However, with deeper insights into their huge potential, novel and effective MSC-based therapies for male infertility will come into being in the near future.


Asunto(s)
Humanos , Masculino , Infertilidad Masculina , Terapéutica , Células Madre Mesenquimatosas , Ingeniería de Tejidos
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