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Inhibition of CatSper and Hv1 Channels and NOX5 Enzyme Affect Progesterone-Induced Increase of Intracellular Calcium Concentration and ROS Generation in Human Sperm | ||
Iranian Journal of Medical Sciences | ||
مقاله 5، دوره 44، شماره 2، خرداد 2019، صفحه 127-134 اصل مقاله (720.98 K) | ||
نوع مقاله: Original Article(s) | ||
شناسه دیجیتال (DOI): 10.30476/ijms.2019.44513 | ||
نویسندگان | ||
Hamideh Ghanbari1؛ Sara Keshtgar* 1؛ Hamid Reza Zare2؛ Behrouz Gharesi-Fard3 | ||
1Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran | ||
2Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran | ||
3Department of Immunology, Infertility Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran | ||
چکیده | ||
Background: Normal sperm function depends on appropriate intracellular calcium (Cai2+) and reactive oxygen species (ROS) levels. Calcium activates NADPH oxidase-5 (NOX5) that leads to ROS generation. The calcium channel of sperm (CatSper) is activated by progesterone and intracellular alkalization. Herein, the interactive role of CatSper, Hv1 channels, and NOX5 enzyme on Cai2+ and ROS generation in human sperm is investigated. Methods: The present laboratory in vitro study was carried out in the School of Medicine, Shiraz University of Medical Sciences (Shiraz, Iran) during 2016. Normal semen samples (n=15) were washed and diluted to 20×106 sperm/mL. The diluted samples were divided into 16 groups containing Ham’s F-10 (the control group), 2 µM NNC (CatSper inhibitor), 1 mM ZnCl2 (Hv1 inhibitor), 1 µM DPI (NOX5 inhibitor), NNC+Zn, NNC+DPI, and NNC+Zn+DPI. The other 8 groups were the same as the above except that they contained 1 µM progesterone. Cell viability and Cai2+ were analyzed by flou-3 AM probe and PI staining, respectively, using flow cytometric method. ROS generation was assessed by chemiluminescence method. Statistical analysis was performed using the one-way ANOVA followed by Tukey’s test. P values | ||
کلیدواژهها | ||
CATSPERB protein, human؛ Hv1 channel, human؛ NADPH oxidase 5؛ Progesterone؛ Spermatozoa | ||
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