Effects of Nicotine and Tocotrienol-Rich Fraction Supplementation on Cytoskeletal Structures of Murine Pre-Implantation Embryos

Abstract

BackgroundCytoskeletal structures, in particular actin and tubulin, provide a fundamental framework in all cells, including embryos. The objective of this study was to evaluate the effects of nicotine, which is a source of oxidative stress, and subsequent supplementation with Tocotrienol-rich fraction (TRF) on actin and tubulin of 2- and 8-cell murine embryos.Material/MethodsThirty female Balb/C mice were divided into 4 groups: Group 1 received: subcutaneous (sc) injection of 0.9% NaCl; Group 2 received sc injection of 3.0 nicotine mg/kg bw/day; Group 3 received 3.0 sc injection of nicotine mg/kg bw/day +60 mg/kg bw/day TRF; and Group 4 received 60 sc injection of TRF mg/kg bw/day for 7 consecutive days. The animals were superovulated with 5 IU PMSG followed by 5 IU hCG 48 h later. Animals were cohabited with fertile males overnight and euthanized through cervical dislocation at 24 h post coitum. Embryos at the 2- and 8-cell stages were harvested, fixed, and stained to visualize actin and tubulin distributions by using CLSM.ResultsResults showed that at 2-cell stage, actin intensities were significantly reduced in the nicotine group compared to that of the control group (p<0.001). In Group 3, the intensity of actin significantly increased compared to that of the nicotine group (p<0.001). At 8-cell stage, actin intensity of the nicotine group was significantly lower than that of the control group (p<0.001). The intensities of actin in Group 3 were increased compared to that of nicotine treatment alone (p<0.001). The same trend was seen in tubulin at 2- and 8-cell stages. Interestingly, both actin and tubulin structures in the TRF-treated groups were enhanced compared to the control.ConclusionsThis study suggests that TRF prevents the deleterious effects of nicotine on the cytoskeletal structures of 2- and 8-cell stages of pre-implantation mice embryos in vitro.

Tocotrienol-Rich Fraction Modulate the Phosphoinositide 3-Kinases/Akt Signaling Pathway Genes and Prevent Oxidative Stress in Nicotine-Induced Pre-Implantation Embryos

“…Nic as an OS inducer has been well established in the previous studies [14,27,40,41]. This study has further confirmed the effects of Nic as an OS inducer, as shown in our data on the increase of OS markers level in the Nic group.…”

“…This data showed the detrimental effects on Nic whereby it these genes which involve in PI3K/Akt pathway may affect the continuity of the cell and nucleus division and may eventually disrupt the development of the preimplantation embryos. We have also documented previously that the actin and tubulin arrangement was also disrupted by Nic in the pre-implantation embryos, which in turn, affecting the embryonic development [41]. Disruption of the cytoskeletal arrangement could be due to the modulation of the PI3K/Akt genes, as shown in this study.…”

“…This study has further shown a reduction of plasma LP accompanied by restoration of enzymatic antioxidants in TRF supplemented group showing the potency of TRF in combating Nicinduced OS. The ability of TRF to effectively prevent the effects of Nic in 2-and 8-cell embryos was indicated by the high intensities of actin and tubulin as previously reported by our group [41]. Approximately 75% of TRF is responsible for the observed benefits, and the presence of three trans double bonds in TRF makes it more readily transferred and incorporated into the cell membranes, which may aid in the repair of actin and tubulin structures.…”

Tocotrienol-Rich Fraction (TRF) Improves the Viability of Wild-Type Saccharomyces cerevisiae in the Initial Stationary Phase

“…Nevertheless, over the years, TRF was documented to exhibit many benefits. In one of the more recent studies, TRF was able to restore actin in cells of murine pre-implantation embryos, after been exposed to oxidative stress triggered by nicotine (Hamirah et al 2017). It also has healing properties; such as protecting astrocytes against glutamate toxicity (Abedi et al 2017), and reducing UV-induced skin redness (Yap 2018).…”