Objective

2

To study the effect and machanism of

Jisuikang

（脊髓康） combined with SRY transcription box factor 2 （SOX-2） on spinal cordinjury （SCI）.

Methods

2

Adult SD rats were randomly divided into blank group and medication group with five rats in each. Rats in the medication group were given 20 g/（kg·d） of

Jisuikang

by gavage， while those in the control group received 18 ml/（kg·d） of normal saline by gavage， both for three days. After that， blood was collected from the abdominal aorta to prepare

Jisuikang

-containing serum and blank serum. Three generations of astrocytes were cultured in vitro in the cerebral cortex of 1~4 day newborn rats， and were divided into three groups including blank serum group， SOX-2 group， and combination of

Jisuikang

-containing serum plus SOX-2 group （SOX-2+JSK group）. Add 10 mmol/L of SOX-2 to the SOX-2 group， and 2.5%

Jisuikang

-containing serum plus 10 mmol/L of SOX-2 to the SOX-2 + JSK group， making the final serum concentration in the medium to 10%； blank serum group was added with 10% of blank serum. After 21 days of induction and culture， the morphology of neuron-like cells in the three groups were observed under light microscope. The expression of neuronal axon marker microtubule-associated protein 2 （MAP2） was detected by cellular immunofluorescence， and the expression of downstream metabolites S-adenosylmethionine （SAM） and acetyl-CoA （acetyl-CoA） were detected by Western Blot. Quantitative PCR analysis was performed for the glycolysis-related genes expression including hexokinase 1 （HK1）， hexokinase 2 （HK2）， phosphofructokinase 1 （PFK1）， phosphofructokinase 2 （PFK2）， and M2-type pyruvate kinase isozyme （PKM2）. The cell energy metabolism analyzer was used to compare the extracellular acidification rate （EACR） and oxygen consumption rate （OCR） of the SOX-2 group and the SOX-2+JSK serum group.

Results

2

Under the light microscope， a large number of neuron-like cells with bipolar and long axons were found in the SOX-2+JSK group， which was significantly more than that in the SOX-2 group； in the blank serum group， astrocytes were found， but with no neuron-like cells. The expression of MAP2， SAM and acetyl-CoA significantly increased in the SOX-2+JSK group compared to those in the blank serum group and the SOX-2 group， while the gene expressions of HK1， HK2， PFK1， PFK2， and PKM2 significantly increased （

P

＜0.05 or

P

＜0.01）. Higher ECAR and OCR were found in the SOX-2+JSK group than the SOX-2 group. There was no significant difference in the expressions of MAP2， SAM， and acetyl-CoA between the blank serum group and the SOX-2 group （

P

＞0.05）.

Conclusion

2

Iisuikang

-containing serum combined with SOX-2 can induce astrocyte reprogramming into neurons， and the machnism of astrocyte reprogramming may be improved through glycolysis.