당뇨가 유발된 햄스터 창상치유에 미치는 벼세포 유래 GM-CSF의 효과
Rice Cell Origin Recombinant Human Granulocyte Macrophage Colony-Stimulating Factor (rrhGM-CSF) Could Improve the Wound Healing in Diabetic Hamster
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상처치유는 growth factor와 cytokine들의 복합적인 작용에 의해 일어나는 과정이다. GM-CSF는 호중구와 대식세포를 분화시키고 각질세포를 증식시키는 다면발현성 사이토카인으로 여러 세포에서 재조합되어 사용되고 있다. (주)핸슨바이오텍에서 개발한 rice cell 유래의 재조합 rrhGM-CSF는 다른 세포에서 유래된 GM-CSF들보 다 탄수화물 측쇄가 많은 것이 특징이다. rrhGM-CSF의 창상치유 효과를 알아보기 위하여 햄스터에 만성 당뇨를 유도하여 상처치유 가 어려운 동물모델을 만든 후, 등쪽에 인위적으로 피부 전 층을 제거하여 창상을 유발하였다. 실험군에는 rrhGM-CSF가 포함된 연 고를 적용하고, 대조군에는 base cream만을 적용하여 재상피화되는 과정과 피부조직의 구조 및 PCNA 반응을 통한 세포증식 정도를 비교하였다. 육안 및 조직학적 관찰 결과, rrhGM-CSF를 적용한 그 룹의 재상피화 속도가 더 빨랐으며, PCNA 면역조직화학적 조사에 서도 증식하는 각질세포의 수가 대조군보다 상대적으로 많은 것으 로 나타남으로써, rrhGM-CSF에 의한 피부상처 치유 효과를 관찰 할 수 있었다. 이러한 결과들로 미루어 보아 벼세포 유래의 rrhGM-CSF는 국소적인 피부 창상 치유에 효과적인 것으로 보이 며, rrhGM-CSF의 풍부한 당화 사슬은 이 단백질이 상처부위에 안 정된 상태로 기능을 나타냄으로써 효과를 오랫동안 지속시킬 수 있었던 것으로 사료된다.
Abstract
GM-CSF is a multipotent growth factor, which also plays an important role during the process of wound healing. rrhGMCSF was specifically produced from rice cell culture in our laboratory (Hanson Biotech Co., Ltd, Daejeon). The rrhGMCSF contains more oligosaccharide side chains than any other types of GM-CSF. This work was taken to evaluate the influence on wound healing of rrhGM-CSF in male golden hamsters. Full thickness skin defects of 9 mm in diameter were made in the back of hamsters, and 100 μL ointment containing rrhGM-CSF 50 μg/mL was applied. Control groups were given ointment without rrhGM-CSF. The wound sizes were relatively reduced and skin was well regenerated in the experimental group compared with the control group. Structurally, reepithelialization and architecture of the skin following injury were well accomplished in the experimental group. And also, positive reaction of PCNA of the skin following injury was more prominent in rrhGM-CSF containing ointment treatment group. Since this type of GM-CSF has highly glycosylated side chains, the effectiveness might be retain longer and stable, regarding acceleration of wound healing in the animal model. The present study has important implications for further development of the therapeutic manipulation of wound healing using rrhGM-CSF.
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