ROS mediates regulation of intracellular Ca2+ induced by angiotensin II in primarily cultured medullary neurons |
| |
Authors: | LIU Xiao-ni CAO Dong-qing ZHANG Na-na TAO Ran DING Ying-jiong JIN Hui-ming LU Ning |
| |
Affiliation: | 1. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China;2. Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, China |
| |
Abstract: | AIM: To investigate the role of reactive oxygen species (ROS) in the regulation of intracellular Ca2+ induced by angiotensin II (Ang II) in the primarily cultured medullary neurons. METHODS: Primarily cultured medullary neurons were prepared from 14-day-old embryos of Sprague-Dawley rats in the study. The identification of medullary neurons was assessed by double-labeling immunofluorescence. To explore the role of ROS, mainly the superoxide (O2·), the O2·generation was measured using the fluorogenic probe dihydroethidium (DHE). To determine intracellular free calcium concentration ([Ca2+]i), the neurons were loaded with the Ca2+-specific dye Fura-2/AM. The cell viability after adding Ang II was also examined using CCK-8 assay. RESULTS: Most of the cultured cells were medullary neurons, more than 80% of which were glutamate positive neurons. Ang II (5 μmol/L) increased the level of ROS within 10 min in the medullary neurons. Ang II at 5 μmol/L induced a significant[Ca2+]i increase in the medullary neurons, and the effect of Ang II occurred rapidly and reached a peak within 20 min after administration. The level of[Ca2+]i started to decline after washout. The Ca2+ elevation induced by Ang II was significantly decreased by apocynin or TEMPOL. No significant difference in the cell viability between control group and 5 μmol/L Ang II treatment group was observed. CONCLUSION: ROS is involved in the regulation of[Ca2+]i induced by Ang II in the primarily cultured medullary neurons, suggesting a potential intracellular signaling mechanism involved in the Ang II-mediated oxidant regulation of central neural control of blood pressure. |
| |
Keywords: | Medullary neurons Reactive oxygen species Intracellular Ca2+ Angiotensin II |
|
| 点击此处可从《园艺学报》浏览原始摘要信息 |
|
点击此处可从《园艺学报》下载全文 |
|