The Synergistic Effects of Ultrasound Waves and Ethanol Exposure on Intracellular Calcium and Reactive Oxygen Species Rates of Hippocampal Cells During the Embryonic Course in Rats

Objective: Ultrasound (US) is an imaging procedure with various applications, such as checking the fetus during pregnancy. To the best of our knowledge no study have investigated the simultaneous effect of diagnostic US waves and ethanol exposure on intracellular calcium ([Ca2+]i) processes and reactive oxygen species (ROS). Thus, in the current study, we first aimed to investigate the impacts of diagnostic US during fetal life on [Ca2+]i and ROS in rats’ hippocampal cells; then whether the diagnostic US has intensified ethanol effects on these two variables.
Materials and Methods: After mating, adult female Wistar rats were divided into six groups as follows: Control group, ethanol group (5 g/kg), groups that were exposed to diagnostic US at frequencies of 3 and 5 MHz for 10 minutes on the 11-13 days of pregnancy, and groups that underwent US waves accompanied with ethanol on the same term of pregnancy. Fourteen days after delivery, the offspring were anesthetized, the hippocampus was isolated, and [Ca2+]i was measured using the Flou 3-AM and fluorimetry instrument after cell culture.
Results: The results showed that the diagnostic US with 3 and 5 MHz frequencies significantly increased [Ca2+]i levels up to 17.8% and 21.8 %, as well as ROS levels by 81.18% and 128.51% compared to the control group. In addition, [Ca2+]i levels in the groups with simultaneous exposure to ethanol and US with frequencies 3 and 5 MHz were 19.19% and 23.33% higher than the ethanol group. Besides, the amount of ROS in the groups with simultaneous exposure to ethanol and US with frequencies 3 and 5 MHz were 77.5% and 172.5% higher than the ethanol group, respectively. However, ethanol alone did not alter intracellular [Ca2+]i and ROS levels.
Conclusion: It is concluded that diagnostic US with 3 and 5 MHz frequencies increase the [Ca2+]i and ROS rate, and it also intensifies the effects of ethanol on the [Ca2+]i and ROS levels.