Gene expression of growth associated protein 43 in activated Schwann cells / 中华骨科杂志

ABSTRACT

Objective To compare activated and normal Schwann cells in GAP43 gene expression. Methods 10 male SD rats, weighed from 100 g to 120 g. The right median nerve of SD rats was transected at the axillary level and was buried in muscle for predegeneration. 1 week later, the distal segment of the transected right median nerve with 1 cm long was harvested. The untreated left median nerve was harvested as control with same length. The epineurium of nerve was stripped, then the nerve tract was cut to small pieces. Schwann cells were obtained by way of double kinases digestion with 0.25% trypsin and 0.03% collegenase. The right median nerve was activated with additional liquid during digestion so as to obtain the activated Schwann cells. The normal Schwann cells were harvested from left median nerve. rt-PCR was used for GAP43 gene enlargement. mRNA was distilled from activated Schwann cells and untreated Schwann cells respectively. Then the mRNA was reversely transcripted to cDNA with SuperScriptTM, and cDNA worked as template for PCR enlargement. The product of PCR was separated with 1% agarose gel electrophoresis for 40 -50 min and stained with SYBR Green Ⅰnucleic acid gel. Fluorescence intensity of GAP43 PCR products was measured and then compared between the experiment group and control group. Results The Fluorescence intensity of GAP43 PCR product of activated Schwann cells was higher than that of normal Schwann cell. There was significant difference (P=0.003, Paired t test). It indicated that GAP43 mRNA of activated Schwann cells was much more than that of the normal Schwann cells. Conclusion GAP43 gene expression is up regulated in activated Schwann cells in contrast to normal Schwann cells. Activated Schwann cells secreting more GAP43, which may be one of the important mechanisms in promoting nerve regeneration.