| Background Drinking has become a worldwide public health problem, and cause many adverse effects to the society, While women drinking also has become more and more common phenomenon. Maternal drinking can seriously affect the growth and development of the fetus during pregnancy, such as cause intrauterine hypoxia, deformity, miscarriage, premature birth, low birth weight and fetal alcohol syndrome (FAS), which is characterized by the central nervous system abnormalities, congenital abnormal heart development, special face, growth retardation, etc. Stable and effective breathing is the precondition of the body’s life. The medial area of nucleus retrofacialis (mNRF) in medulla oblongata respiratory center is the key part of basic rhythmic respiration origin. Central respiratory system diseases, especially the pathological change of medullary respiratory center, become common diseases and frequently encountered disease in the clinical work. In addition, there are more and more abnormal central respiratory induced by alcoholism. Therefore, it has a broad application prospect and significance to study the effect and mechanism of pranatal alcohol exposure on medulla oblongata respiratory center of offspring for the prevention and treatment of related diseases.Objective To studies the effect of 5-HT2A receptors in prenatal alcohol exposure inhibit medullary slice rhythmic respiratory discharge activities (RRDA) in neonatal rat.Methods1. Clean level Sprague-Dawley adult rats, male and female ratio were 2:1, female and male mice were randomly divided into six groups:control group and 1% prenatal alcohol exposure group,2% prenatal alcohol exposure group,4% prenatal alcohol exposure group,8% prenatal alcohol exposure group,10% prenatal alcohol exposure group (v/v). The male and female rats were 2:1 cage mate. Control group rats were given water to drink freely, and different concentration alcohol exposure group rats were given corresponding concentration alcohol aqueous solution as the only drinking water (freely) from mating before lw to postpartum 3d.2d neonatal male and female rats were used in this experiment. In vitro medulla slices in neonatal rats were made and using neurophysiological techniques to compare medulla slices RRDA of neonatal rats in each group, and the change of inspiratory time (TI), and integral amplitude (IA), respiratory frequency (RF) to clear the optimal alcohol concentration of prenatal alcohol exposure model rats.2. To study whether 5-HT2A receptors in prenatal alcohol exposure inhibition medullary slice rhythmic respiratory discharge activities (RRDA) in neonatal rat, the experiment is further divided into 6 groups:control group, prenatal alcohol exposure group, control DOI group, prenatal alcohol exposure DOI group, control ketanserine group, prenatal alcohol exposure ketanserine group.3. Using Western Blot technology to research 5-HT2A receptor protein level in the respiratory center mNRF respiratory neuron of prenatal alcohol exposure neonatal rat.4. Using qRT-PCR technology to research 5-HT2A receptor mRNA level in the respiratory center mNRF respiratory neuron of prenatal alcohol exposure neonatal rat.Results1. Control group and 1%～8% prenatal alcohol exposure groups slices RRDA were stability without attenuation in 60min, and the experiment model were stable and reliable. Compared with control group, RRDA in 1% and 2% alcohol group did not change significantly (P>0.05); RRDA gradually slowed down with the increase of alcohol concentration in 4%～10% alcohol group, shortened TI, reduced IA, decreased RF (P<0.05); RRDA of 10% alcohol exposure group was weaker than that of 8% alcohol exposure group, and the RRDA became irregular, so we chose 8% alcohol as the optimum concentration of prenatal alcohol exposure.2.5-HT2A receptor specificity agonist DOI could excite slices discharge of the two groups, and the prenatal alcohol exposure group excited effect was weaker than the control group.5-HT2A receptor specificity antagonist ketanserine had inhibitory effect on slices discharge of the two groups, while the prenatal alcohol exposure group inhibitory effect was weaker than the experimental group.3.5-HT2A receptor protein level gradually slowed down with the increase of alcohol concentration in the respiratory center mNRF respiratory neuron of 4% and 8% prenatal alcohol exposure group (P＜0.05).4. qRT-PCR results show that,8% prenatal alcohol expoure down regulated 5-HT2A receptor mRNA level in the respiratory center mNRF respiratory neuron in neonatal rat.Conclusions1. Prenatal alcohol exposure depresses RRDA in the medullaly slice of neonatal rat, and inhibits the adjustment effect of 5-HT2A receptor to RRDA.2. Prenatal alcohol exposure down-regulate the 5-HT2A receptor protein and mRNA level, which is one of the reason prenatal alcohol exposure inhibit RRDA. |
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