척수 편측절단 손상 후 일회성 운동량에 따른 neurotrophins 단백질발현 및 수용체 활성
The effects of quantity of treadmill exercise on expression of neurotrophins and receptor after spinal cord hemisection in the rats.
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본 연구는 흰쥐의 흉추 9번을 반측절단 손상을 유발시키고, 1주 동안의 휴식 후 저강도 운동 처치를 15분, 30분, 60분, 90분으로 나누어 1회 실시하였으며, 운동직후 손상신경조직을 적출하여 형태적 변화와 신경성장관련 인자들의 발현정도를 분석하였다. 실험동물은 7주령된 SD계열 수컷 흰쥐 60마리를 실험대상으로 normal 그룹, 척수손상(SCI) 대조군, SCI+운동15분그룹, SCI+운동30분그룹, SCI+운동60분그룹, SCI+운동90분그룹으로 총 6그룹으로 설정하였다. 운동처치는 저강도운동으로 1회 실시하였다. 그 결과 BDNF 단백질의 발현량은 운동 60분 > 30분 > 90분 > 15분순으로 60분에서 가장 많이 발현되었으며, 수용체인 TrkB mRNA의 발현량은 30분 운동그룹에서만 높게 발현되었다. NGF 단백질 발현량은 손상 후 운동그룹 모두가 손상그룹보다 높게 나타났으며, 수용체인 TrkA mRNA의 발현량은 운동 15분 > 60분 > 30분, 90분순으로 운동 15분에 가장 높게 발현되었다. NT3 단백질 발현량에서는 운동 30분, 60분 > 90분 > 15분순으로 운동 30분과 60분에서 높게 나타났으며, 수용체인 TrkC mRNA의 발현량은 운동 30분 > 60분, 90분 > 15분순으로 운동 30분에서 가장 높게 발현되었다. 형태학적인 변화로 반응성 별아교세포의 변화는 일회적인 운동량만으로는 변화가 나타나지 않는 것을 확인하였다. 이상의 결과를 종합한 결론은 척수손상 후 운동처치에 따른 신경성장관련 인자들의 발현량은 운동 약 30분~60분정도에서 높게 나타나는 것을 확인하였다.
Abstract
Sixty male Sprague-Dawley rats (230±10 g; 7 week in age) were assigned equally to six different groups; Normal (n=10), Spinal cord injury (SCI)(n=10), SCI+15m treadmill training (TMT)(n=10), SCI+30 mTMT (n=10), SCI+60 mTMT (n=10), SCI+90 mTMT (n=10). Every rat in SCI and SCI+TMT groups underwent laminectomy at thoracic vertebra ninth(T9) level and then hemisection the exposed spinal cord site in the anesthetized condition. After one week-recovery from injury, rats in SCI+TMT group exercised on a motorized treadmill for 15, 30, 60, 90 min/d. All tissues were used for H&E, immunofluorescence staining, Western blot and RT-PCR. The data obtained were analyzed by one-way ANOVA and Tukey`s post-hoc. and presented as mean±standard deviation. BDNF induction levels in the injured spinal cord significantly showed more increases in exercise group with SCI and were highly correlated with time of exercise, but NGF did not differ from another groups. TrkB mRNA which is receptor for BDNF was increased in only exercise group for 30 minutes and TrkA mRNA which is receptor for NGF was upregulated in two exercise group for 15 and 60 minutes. Also, TrkC mRNA, activated by NT-3, was significantly enhanced in exercise group for 30 minutes. Thus present data suggest that exercise plays a important role for axonal regeneration and functional recovery after SCI.
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