內皮唾酸蛋白(CD248)是第一型跨模糖蛋白，會大量表達在周細胞以及纖維母細胞上。真皮纖維母細胞在傷口癒合過程的增生期(proliferative)及重塑期(remodeling)具有很重要的角色。本實驗室先前的研究發現，小鼠在創傷後的第7到14天，傷口部位的CD248表達明顯上升。傷口癒合的速度在CD248基因剔除的小鼠顯著變慢，糖尿病小鼠的傷口癒合速度亦變慢。本研究主要目標是探討CD248是否在糖尿病傷口癒合扮演角色。本研究發現與正常小鼠相比，第一型或第二型糖尿病小鼠的傷口中，CD248的表現量在創傷後的7到14天明顯下降。在體外細胞實驗中發現在高葡萄糖濃度(25 mM)的培養下人類真皮纖維母細胞(normal human dermal fibroblasts)NHDFs的CD248蛋白和基因表現量下降，同時胰島素受體(insulin receptor, INSR)的蛋白表現也下降。根據文獻，高葡萄糖環境會引起細胞衰老。因此在研究利用bleomycin誘導NHDFs衰老後，發現CD248的蛋白和基因表現量下降，INSR的蛋白表現也下降，而當CD248基因被剔除後NHDFs之衰老加劇，胰島素所誘導之訊息傳遞以及生物功能也受到抑制。此外，我們透過免疫螢光共軛焦證實CD248與INSR共位存在於NHDFs之次細胞胞器，並藉由免疫共沉澱發現CD248與INSR相結合，因此我們推測高葡萄糖經由抑制CD248基因表現而造成細胞衰老及INSR表達。利用小鼠進行活體內研究，發現若將小鼠的CD248基因剔除，會阻礙胰島素治療之傷口修復過程。總結以上的結論，在糖尿病傷口中，CD248的表達下降可能是導致傷口不易癒合的原因之一，在傷口癒合之過程中CD248通過調節纖維母細胞的細胞衰老和胰島素誘導之反應，是傷口癒合主要的關鍵因素之一。

CD248 is a type 1 transmembrane glycoprotein which is specifically expressed in pericytes and fibroblasts in mice after birth. Dermal fibroblasts play a critical role in cutaneous wound healing, especially in proliferative and remodeling phases. In the previous studies, we demonstrated that CD248 expression was increased in the wound region in normal mice between day 7-14 after wound. The wound healing was significantly compromised in the CD248 knockout mice. However, the temporal pattern of CD248 expression and its function in diabetic wound remain unexplored. Here, we demonstrated that CD248 expression was significantly down-regulated on day 7-14 in type 1 or 2 diabetic mice. The mRNA and protein levels of CD248 expression and insulin receptor (INSR) protein were decreased in normal human dermal fibroblasts (NHDFs) cultured in the high glucose (25mM) medium. It has been known that high glucose can induce cell senescence. The bleomycin-mediated senescent dermal fibroblasts displayed a decrease in the mRNA and protein levels of CD248 expression and insulin receptor protein. Furthermore, CD248 silencing facilitated cellular senescence. We noticed that when CD248 expression was silenced, the insulin-induced signaling transduction and biological functions were compromised. Moreover, CD248 and insulin receptor were co-localized in sub-cellular organelles in fibroblasts and the association of CD248 and insulin receptor was demonstrated by co-immunoprecipitation. CD248 knockout retarded insulin-enhanced wound healing process in mice. In summary, wound-induced CD248 expression can be mitigated in the diabetic situation and CD248 acts as a key factor for wound healing by regulating cellular senescence and insulin-induced responses in fibroblasts.

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