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Proliferative and oxidative response of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats exposed to ketogenic diet

Wójcik, M., Wessely-Szponder, J., Kosior-Korzecka, U.
Polish journal of veterinary sciences 2015 v.17 no.4 pp. 703-711
centrifugation, collagenase, free radicals, hepatocytes, ketogenic diet, liver, nitric oxide, rats, superoxide anion
Ketogenic diet (KD) is considered in the context of its anti-epileptic effects, but its influence on liver dysfunction has not been elucidated yet. The study was aimed to investigate the activity of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats fed with KD, in respect of NO and superoxide generation by these cells as well as their proliferative activity in vitro. We also sought to characterize the plasma FFA profiles in control and ketogenic rats. Hep and HSC were isolated by the collagenase perfusion method and separated by the Percoll gradient centrifugation. After the 4ᵗʰ, 8ᵗʰ and 12ᵗʰ day of incubation, the media were collected for further analysis. NO generation increased within the time of incubation both in Hep and HSC isolated from KD-rats. In HSC group NO production raised significantly from 2.65 ± 0.07 μM/106 cells on 4ᵗʰ day of incubation to 5.49 ± 1.2 μM/106 cells on 12ᵗʰ day of incubation. In respect to O₂⁻. generation experimental Hep and HSC provide considerably higher quantities of this free radical. until 12ᵗʰ day of incubation (2.5 ± 0.07 and 3.2 ± 0.3 nM/106 cells, respectively). Although KD exerts anti-proliferative effect on hepatocytes, in respect to HSC it intensifies their proliferative activity. Furthermore, as we estimated on the basis of NO and O₂⁻. generation both Hep and HSC exposed to KD are the source of free radicals