The Study Of Resveratrol Emulsomes

When pregnant women drink excessive alcohol during pregnancy, it may cause abortion and stillbirthas alcohol can travel through the placental barriers along with blood, as well as resulting in some irreversibleinfluences on the fetus after birth, such as growth disorders, facial deformity, and especially nerve systemdevelopmental damages (Fetal Alcohol Syndrome, FAS). FAS symptoms may optic ones such as strabism,microphthalmia, congenital myopia, and amblyopia, which are often related to retinal abnormalities..In thisstudy, our aim is to use C5B/6J mice to construct prenatal alcohol exposure (PAE) model for theinvestigation of its effects on retinal teratology, cell development and apoptosis. Our study would providetheoretical basis for the etiology of ophthalmic diseases caused by PAE.Objective: Our objective is to investigate the normal development and cell differentiation process ofretina, as well as to elucidate the teratological effects of PAE on retinal cell development and apoptosis.Methods: PAE model was constructed using C57BL/6J mice, and P0, P7, P14, and P30filial micewere selected to observe the normal development and cell differentiation process of retina using HEstaining and immunofluorescent labeling techniques, as well as to study the teratological effects of alcoholon retinal cell development and apoptosis.Results:1. The development of the retinal general structure: the formation of retinal lamination and thedevelopment of horizontal and bipolar cells:(1) Formation of Retinal Lamination:There were only four layers in P0retina, including the ganglion cell layer, inner plexiform layer,neuroblastoma cell layer, and purpurogenous membrane. At P7the outer plexiform layer appeared anddivided the neuroblastoma cell layer into inner and outer granular layers, after which the outer plexiformlayer widened and the layer of rods and cones appeared. At P14(opening of the pup eye lids) the retinalten-layered structure was completely formed. The results showed that the development of retinalmorphology was consistent with the consummation of its functions.(2) The development of horizontal cells and bipolar cells:In this study, we selected Doublecortin and PKC-α to mark the horizontal cells and bipolar cells.Horizontal cell development: the Doublecortin positive cells appeared at P0and were located in the neuroblastoma cell layer near the purpurogenous membrane, after which they started to concentratedownwards. At P7the Doublecortin positive cells reached the target layer, with their dendrites stretchinginto the outer plexiform layer, forming a “T” structure with the cell bodies. Afterwards, the number ofdendrites stretching into the outer plexiform layer gradually increased, until a fan-shaped structure wasformed by the dendrites and cell bodies, indicating that the development of horizontal cells was basicallycomplete at this time. Bipolar cell development: At P0there was no PKC-α-positive cell, which did notcome out until P7. At this time, the cell bodies showed multi-layer arrangement in the inner nuclear layerclose to the outer plexiform layer and established connections with the axons of ganglion cells. However,the dendritic plexuses stretching into the outer plexiform layer were not obvious. Afterwards, the number ofPKC-α positive cells increased and were arranged into a single tightly-packed layer at P14, with increasedaxonal densities. A large number of plexuses could be seen in the outer plexiform layer, showing the typicalbipolar cells morphology at this time.2. The effects of prenatal alcohol exposure (PAE) on retinal lamination and the development of horizontaland bipolar cells(1) The effects PAE on retinal lamination:In this study, we found that the lamination of PAE mice retina was thickened and delayed afterbirth in a dose-dependent and long-term manner.(2) The effects of PAE on the development of horizontal and bipolar cells:At each age point, the number of retina horizontal cells and bipolar cells in PAE group wasreduced in a dose-dependent and long-term manner. In addition, the polarity of horizontal cells in PAEgroup was disordered in a dose-dependent manner.3. The teratological effects of PAE on retina and cell apoptosis:By performing HE staining on each dosage group at P0, we found that deformed retina did notappear in the control and low-dose group, while the teratology rate of high-dose group was16%. Themanifestation of deformity was mainly the proliferation of retinal cells, resulting in thickening and foldingof the retina. In addition, activated caspase-3, caspase-8and caspase-9TUNEL were used to mark theapoptotic cells. The statistical results showed that under physiological conditions, the retinal ganglion cellsand inner granular layer cells underwent apoptosis via the death receptor pathway and mitochondrial pathway, with the former pathway playing the major role. During prenatal alcohol exposure, alcoholinduced the apoptosis of retinal cells via the death receptor and mitochondrial pathways in addition to thephysiological apoptotic pathway in a dose-dependent and long-term effect.In summary, PAE could cause the developmental retardation, decrease in functional neurons,morphological deformity, increased rate in retinal teratology, and the increase in cellular apoptosis in filialpup retina. These could be the pathological basis causing FAS related ophthalmic diseases.