Influence Of Electro-acupuncture On The Expression Of NeuroD In Rats’ Brain With Intrauterine Distress

Objective A rat model was established with delayed cesarean section, mimicking neonatal hypoxic to study the influence of electro-acupuncture on the expression of Neuro D in brain tissue. Exploring its mechanism of action, to support clinical application of electroacupuncture with basic experimental basis in hypoxic-ischemic encephalopat-hy.Methods1.With delay cesarean section, intrauterine fetal distress of the rat model was established. Neural behavior experiment(elevated plus maze, open field test, morris water maze) and frozen section HE staining were used to observe the effects of intrauterine asphyxia time of the newborn rats.2.When these newborn rats were three days old, they were performed electro-acupuncture treatment, fifteen minutes every day for continuous seven days;Neuronal markers(Neu N and Neuro D) were used in immunofluo-rescence staining, to observe the Neuro D expression after EA treatment.3. Using real-time PCR analyzed the relative expression of Neuro D when newborn rats were Day16, Day22, Day35; Using protein immunoblot(Western blot) detected the relative expression of Neuro D when newborn rats were Day16, Day22, Day35;Using immunofluorescence double labelled staining(MAP-2 and DAPI, Neu N and DAPI),and three labelled staining(Neuro D、MAP- 2 and DAPI, Neuro D、Neu N and DAPI),laser confocal microscopy observe Neuro D expression intensity when newborn rats were Day22.Results1.With the increase intrauterine distress time of fetal rats, newborn rats’ neural abnormal behaviors were more and more obvious, and fatal convulsions even happened when hypoxic time was up to thirty minutes. Various behavioral test(elevated plus maze, open field test, Morris water maze) results showed that the longer hypoxic time,the lower activity level, the higher anxiety level, the lesser performs at spontaneous activity and exploratory behavior, and learning and memory ability changed. In hypoxic ten minutes and twenty minutes groups, learning ability gradually improved but memory ability kept unchangable, and in hypoxic thirty minutes group, learning and memory abilities are lower than the blank control group.2. Immunofluorescence labelled staining Neu N and Neuro D showed that EA intervention triggered the Neuro D overexpression, and Neu N also had the same results,the max expression was in the hypoxic twenty minutes.3. The real-time PCR and Western-blot results showed that the hypoxic twenty minutes group had the highest relative expression level of Neuro D,immunofluorescence double labelled staining and the three labelled staining results also have the same trend.Conclusions The hypoxic twenty minutes group achieved the best effect after EA treatment was performed. EA intervention could stimulate Neuro D transcription and translation to overexpress, to induce NSCs differentiation to participate in repairing the damaged nerve tissue for making never regeneration and functions reconstruction happened. And after a period of time at the end of the EA intervention, the stimulus effects still exist in groups.