| Summary: | 碩士 === 國防醫學院 === 生物化學研究所 === 107 === The type II diabetic patients have poor peripheral blood circulation and vascular and nerve atrophy, which induce hypoxia condition and non-effective immune response. These effects may trigger the formation of chronic wound. Previous results have shown matriptase and prostasin are important in chronic wound healing. Therefore, we would like to learn the expression of matriptase and prostasin in different clinical outcome of chronic diabetic ulcers and compare the differences between foot and non-foot area.
At first, compared with DM foot ulcer, the expression extent of matriptase and prostasin in DM non-foot ulcer was similar with normal skin, but the expression intensity of HAI-1 was still lower like DM foot ulcer. Secondly, although the expression of matriptase and prostasin are different between better outcome and worse outcome of diabetic ulcer, however the expression of matriptase could not extend to normal skin, and the expression extent of prostasin also could not withdrawal to the upper spinous layers. Thus, we suggested that the tissues after debridement, in the better outcome of DM ulcer, are still in bad condition. Thirdly, we discovered 45.4 percent DM ulcers have shown activated matriptase on the cell membrane in the upper spinous and granular layers, which is similar with the activated prostasin expression. At the same location, mostly, the expression intensity of HAI-1 was stronger, and highly glycosylated HAI-2 （DC16）also was detected. Further, in immortalized human keratinocytes cell line （HaCaT）, we found a 100 kDa complex activated matriptase and highly glycosylated HAI-2 by using immunoprecipitation. Therefore, we suggested that highly glycosylated HAI-2 was expressed on the cell membrane in the upper spinous and granular layers because they can help HAI-1 to inhibit more activated matriptase. Fourthly, the whole tissues of DM ulcer from the same debridement could expand from the healthy side to the wound side. We found that the expression extent of matriptase at the wound side was lower than the healthy side. At the same time, activated prostasin was spread out more from granular layers to the spinous layers at the wound side. Thus, we recommend that there were some reasons to make the dissimilar distribution of matriptase, prostasin, and their inhibitors between healthy side and wound side. Finally, we used HaCaT cells to study the expression of prostasin under terminal differentiation. We tried to mimic the terminal differentiation condition by increasing calcium concentration or increase cell density. In addition to more cell clustering, we found the expression of matriptase, prostasin, HAI-1, and HAI-2 were increased under differentiation. Hence, we suggested that the differentiation of human keratinocytes did not only influence prostasin, but also induce the expression of other proteins.
In this thesis, we observed the expression of matriptase, prostasin, HAI-1 and HAI-2, and concluded that these proteins are closely related in wound healing at different stages. Therefore, we can find the possible strategy for chronic wound healing, and this information will provide clinical treatment to accelerate chronic wound healing.
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