Objective To observe the effect of acupuncture serum on the expression of microtubule associated protein-2(MAP-2) and nerve growth associated protein-43(GAP-43) in cultured hippocampal neurons of convulsive rats. Methods The acute convulsion model was induced by intraperitoneal injection of pentylenetetrazol in SD rats who were then randomized into model group and acupuncture group. Rats of the acupuncture group received manual acupuncture stimulation of “Baihui”(GV20) and “Dazhui”(GV14) for 30 min

once daily for 7 days. Then

the blood samples taken from the abdominal aorta of rats in the convulsion model and acupuncture groups were processed into serum samples

i.e. non-acupuncture serum and acupuncture se-rum. The primary-cultured hippocampal neurons of fetal rats were cultured for 10 days and then divided into normal extracellular fluid(normal) group

magnesium(Mg

(2+)

) free extracellular fluid group

acupuncture serum group and non-acupuncture serum group. At the 10 th day

the neurons in the normal group were cultured continuously in extracellular fluid for 3 h

and then cultured in DMEM/F12(1∶1) medium(planting fluid); neurons in the Mg(2+)) free extracellular fluid group

acupuncture serum group and non-acupuncture serum group. At the 10 th day

the neurons in the normal group were cultured continuously in extracellular fluid for 3 h

and then cultured in DMEM/F12(1∶1) medium(planting fluid); neurons in the Mg

(2+)

free group were cultured in magnesium-free fluid medium to induce epileptic-like discharge; neurons in the acupuncture serum group were cultured in the mixed medium of planting fluid and 10% acupuncture serum; and neurons in the non-acupuncture serum were cultured in the mixed culture medium of planting fluid and non-acupuncture serum(10%). At last

these neurons in the above-mentioned groups were cultured in the magnesium-free extracellular fluid continuously for 2

12 and 48 h

respectively

followed by detecting the expression levels of MAP-2 and GAP-43 proteins at the 3 time points by using immunofluorescence and Western blot

separately.Results The rate of MAP-2 positive cells and protein expression at 2

12 and 48 h

and the rate of GAP-43 positive cells and protein expression at 12 and 48 h in the hippocampal neurons were significantly down-regulated in the Mg(2+) free group were cultured in magnesium-free fluid medium to induce epileptic-like discharge; neurons in the acupuncture serum group were cultured in the mixed medium of planting fluid and 10% acupuncture serum; and neurons in the non-acupuncture serum were cultured in the mixed culture medium of planting fluid and non-acupuncture serum(10%). At last

these neurons in the above-mentioned groups were cultured in the magnesium-free extracellular fluid continuously for 2

12 and 48 h

respectively

followed by detecting the expression levels of MAP-2 and GAP-43 proteins at the 3 time points by using immunofluorescence and Western blot

separately.Results The rate of MAP-2 positive cells and protein expression at 2

12 and 48 h

and the rate of GAP-43 positive cells and protein expression at 12 and 48 h in the hippocampal neurons were significantly down-regulated in the Mg

(2+)

free group in contrast to the normal group(P

<

0.05

P

<

0.01). Compared to the Mg(2+) free group in contrast to the normal group(P

<

0.05

P

<

0.01). Compared to the Mg

(2+)

free group

the rates of MAP-2 and GAP-43 positive cells and protein expression at 2

12 and 48 h were considerably up-regulated in the acupuncture serum group(P

<

0.05

P

<

0.01)

but not in the non-acupuncture serum group(P

>

0.05).Conclusion Acupuncture serum can significantly up-regulate the expression of MAP-2 and GAP-43 proteins in hip-pocampal neurons

which may play a positive role in improving synaptic plasticity and neuronal damage in convulsion rats.