Effects Of Ethanol On Rat Primary Cortical Astrocytes Expression Of GFAP&Plk2

ObjectiveEthanol is the world’s most widely consumed one substance, which one is situatedin the top fifth in20different kinds of common harming drugs; reverse alcohol-relateddamage is an urgent problem to public health.Brain astrocytes are the first cell barrierfollowing vascular endothelial cells that effected by ethanol and its toxicity, humanlong-term ethanol exposure leads to dendrites and astrocytes irreversible damage, butneuronal cell bodies and most axons showed no irreversible injury, that prompteddendrites and astrocytes in the distal protrusions may be toxic injury key target.As aresult of glial fibrillary acid protein (GFAP) content has regularity response thatincreased with short-term ethanol exposure and decreased with long-term ethanolexposure, and the surface of rats astrocyte cells are more smooth with chronicethanol-treated, and the GFAP is indispensable when the structural requirementsimposed on the ASTs, which suggest that GFAP and actin network regulation existnecessarily related, but at present the GFAP entry actin regulatory pathway mechanismis still unknown. Combined with the former studies to postsynaptic neurons and T cellsin Plk2-SPAR-actin pathway, based on the study of the influence of rat primaryastrocytes activity with different concentration of ethanol exposure, discussed theconnection between the changes of GFAP and the expression of Plk2and SPAR,identified Plk2-SPAR-GFAP pathway which contributes to the understanding of ethanolinjury key cellular targets, and the development of targeted neuroglial and dendriticplasticity of effective interventions, that expected to recover cognitive abilities ofalcohol dependent patients, and provides hope and power for alcohol dependent patientsto stop the ethanol abuse. MethodIn vitro culture of neonatal rat primary cortical astrocytes, using GFAPimmunofluorescence staining method for identification of cell purity, detected withMTT of different concentrations of ethanol exposure on astrocytes activity: a finalconcentration of0mM,50mM,100mM,200mM,400mM,600mM,800mM ethanolmedium were2×104/hole astrocytes24h,48h,72h. By immunofluorescence stainingto observe the effects of different concentrations of ethanol exposure on GFAPexpression influence. Application of Real-time PCR from nucleic acid level detection ofintermediate filament GFAP and Plk2and SPAR mRNA table level.Results1. Neonatal rat primary cortical astrocytes in the purity identification of95±1.64%;2. Different concentration of ethanol on astrocyte activity in newborn ratsIn the ethanol treatment in24h cases, neonatal rat cerebral cortex astrocytesactivity compared with the normal group, the50mM and800mM ethanol treatmentgroup differences (P＜0.05).In the ethanol treatment in48h cases, compared with the normal group,400mM,600mM and800mM ethanol treatment group differences（P＜0.05）, and600mM,800mM group showed significant difference(P＜0.01).In the ethanol treatment in72h cases, compared with the normal group,400mM,600mM and800mM ethanol treatment group difference（sP＜0.05）, and400mM,800mM group showed significant difference（P＜0.01）.On the whole, ethanol for reactive astrocytes effects exist to rise (activity) after afall (decreased activity) trend.3. Different concentration of ethanol on astrocyte Plk2, SPAR and GFAP mRNAexpression in rats.In200mM ethanol treatment, Plk2mRNA at10min was significantly increasedexpression (P <0.05),30min volume reached a peak (P <0.01), then in accordancewith the passage of time, the Plk2expression of mRNA in a time-dependent decreasein6, h and control group have no difference;In200mM ethanol, SPAR mRNA expression at the bottom, with the time process,SPAR mRNA amount begins to pick up,48h, SPAR mRNA expression was increasedto control level;In200mM, GFAP mRNA expression gradually increased, but in24h was significantly higher than those in control group (P <0.01),48h expression increasesfurther,72h and48h expression similarity, maintained at high level.Conclution1. With different ethanol concentration for different exposure time for the stimulusintensity, stimulus duration, when stimulus duration was constant, astrocytes to differentconcentrations of ethanol stimulation response, but the response (cell activity andGFAP expression) expression was two stage effect, i.e. astrocytes to ethanol exposurehas a certain resistance, more than a certain limit, resistance to disappear.2. Plk2-SPAR pathway of astrocytes in200mM ethanol exposure shows atime-dependent changing.3. In200mM ethanol exposure, GFAP mRNA expression event occurs after thePlk2-SPAR event in astrocytes.