Transcriptional level of calmodulin mRNA during the earlier differentiation of rat bone marrow mesenchymal stem cells induced by astragalus mongholicus*,*,☆

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Abstract

BACKGROUND:

Astragalus mongholicus can promote differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons, but induction mechanism was few. At present, the induction may be associated with the ability of anti-oxidation.

OBJECTIVE:

To observe the changes in the transcriptional level of calmodulin mRNA during the earlier differentiation period of rat BMSCs induced by astragalus mongholicus injection.

DESIGN, TIME AND SETTING:

The cytological in vitro controlled study was performed at the Experimental Center of Hebei North University from October to December 2007.

MATERIALS:

One clean male rat aged 6 weeks were purchased from Experimental Animal Center, Chinese Academy of Medical Sciences. Astragalus mongholicus injection (lot number 060105) was purchased from Dali, China.

METHODS:

After isolation, culture and passage, four-passage BMSCs at 4×105/L were incubated in a 12-well plate in DMEM containing 200 g/L astragalus mongholicus injection and 15% fetal bovine serum. BMSCs underwent immunocytochemical staining. At fifth passage, BMSCs were equally assigned into 4 centrifuge tubes. Tube 1 served as control group. BMSCs in other three tubes were treated with above-mentioned medium for 30, 60 and 120 minutes. The harvesting cells were treated with RT-PCR to measure the transcriptional level of camodulin mRNA.

MAIN OUTCOME MEASURES:

Induction of astragalus mongholicus on BMSCs, and transcriptional level of camodulin mRNA after induction were measured.

RESULTS:

After BMSCs are induced by astragalus mongholicus for 5 hours, the morphology of a small number of cells was changed. The cell body was round and the long and thin prominences stretched out from the body. The results of immunocytochemical staining showed that there were some positive staining cells for nestin and neurone specific enolase, a small number of cells presented positive staining for microtubule-associated protein-2, but the degrees of the positive staining was weak comparatively in glial fibrillary acidic protein positive cells. The results of RT-PCR indicated that the gene of calmodulin had been transcribed at the earlier period of BMSC differentiation induced by astragalus mongholicus and the transcriptional level was upregulated gradually with prolonged induction time. Significant differences in relative amount of calmodulin mRNA between the 30, 60, 120 min groups and control groups (F=153.315, P=0.000).

CONCLUSION:

Astragalus mongholicus can promote the differentiation of BMSCs into neurons. During early induction period, the transcriptional level is upregulated. It is indicated that the induction effect may be associated with calmodulin-mediated signal transduction.

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