Objective To explore the effect of electroacupuncture(EA)serum on expression of myogenic differentiation antigen(Myod)and autophagy-related protein Beclin 1 in cultured muscle satellite cells of rats under starvation conditions.MethodsThe primary multifidus muscle satellite cells of one male SD rat were isolated and cultured to obtain the 3

(rd)

generation of cells. The EA serum was got from the rat received EA stimulation of bilateral"Weizhong"(BL40

2 Hz/10 Hz

1 mA

duration of 20 min

once daily for 7 days). The cell suspension(2×10(rd) generation of cells. The EA serum was got from the rat received EA stimulation of bilateral"Weizhong"(BL40

2 Hz/10 Hz

1 mA

duration of 20 min

once daily for 7 days). The cell suspension(2×10

4

/well)of the 34/well)of the 3

(rd)

generation of cultured cells was transferred to each well of a 96-well plate in medium containing 10% fetal bovine serum(FBS).Twelve duplicate wells were set up for the blank control serum(without FBS)

10% FBS

10% EA serum

20% EA serum and 30% EA serum groups and incubated for 12 h and 24 h

respectively. Each well was supplemented with 10 μL CCK-8 reagent to be incubated for 1 h again for observing the state of cell proliferation. After culturing the primary muscle satellite cells in serum-free medium for 12 h

the cells were randomly divided into serum-free group

10% fetal bovine serum group and optimal concentration electroacupuncture serum group

and serum of corresponding concentration was added respectively. The expression levels of Beclin 1 and cell-proliferation-related protein Myod were detected by Western blot.ResultsCCK-8 assay displayed that the proliferation levels were significantly higher at 12 h and 24 h after serum intervention in the 10% FBS

10% EA serum

20% EA serum and 30% EA serum groups than that in the blank control serum group(P

<

0. 01)

and at 24 h in the 3 EA serum groups than in the 10% FBS group(P

<

0. 01)

but without significant difference among the three EA serum groups(P

>

0. 05). As a result

10% EA serum was selected as the optimal concentration for Western blot tests. No significant difference was found in the expression levels of Myod and Beclin 1 proteins among the serum-free

10% FBS and 10% EA serum groups before intervention(P

>

0. 05)

and there was a marked up-regulation of Myod expression and an obvious down-regulation of Beclin 1 expression at 12 h in both the 10%EA serum and 10% FBS groups in comparison with their own pre-intervention(P

<

0. 05). There were a marked up-regulation of Myod expression at both 12 h and 24 h and Beclin 1 expression at 24 h in both the 10% EA serum group and10% FBS group than that in the serum-free group(P

<

0. 05)

and an obvious down-regulation of Beclin 1 expression at12 h in both 10% FBS and 10% EA serum groups than that in the serum-free group(P

<

0. 05

P

<

0. 01). After 24 h's serum intervention

there was an increase of Myod expression and a reduction of Beclin 1 expression in both 10% FBS and 10% EA serum groups compared with those after the 12 h intervention(P

<

0. 05). No significant differences were found between the 10% FBS and 10% EA serum groups in the expression levels of Myod and Beclin 1 proteins(P

>

0. 05).ConclusionEA serum can promote proliferation of cultured muscle satellite cells under starvation conditions

which is related to its functions in regulating expression of Beclin 1 and cell-proliferation-related protein Myod.