黃相碩賴德仁劉幸怡Hsin-Yi醫學研究所2026-10-289999-12-312009https://handle.ncl.edu.tw/11296/5rx3fqhttps://scholars.csmu.edu.tw/handle/123456789/35615演化上，Sonic hedgehog (SHH)是具有高度的保留性，其對脊椎動物器官發育扮演重要的調控角色。除了個體發育上的重要性，SHH的訊息傳遞也可促進損傷組織的血管新生。在器官的缺血及再灌注傷害研究中，發現組織細胞在缺血的狀態下，細胞的SHH表現量會上升，也會間接促進參與血管新生的VEGF、angiopoietin 1和2 (Ang1、Ang2)的表現，進而減低缺血對器官所造成的損傷。在個體中，腦部需要大量的血液運送氧氣和養分來供其運作；因此，腦部是極為容易受到缺血及再灌注傷害的器官之一。先前的研究已發現，SHH可以透過增加缺血區域的血管新生作用，及預防受損細胞凋亡來減緩缺血及再灌注對肌肉組織所帶來的傷害，並且，由SHH誘導產生的細胞激素能夠幫助修復受損的細胞組織，但SHH對腦部的缺血及再灌注傷害，是否具有相似的保護機制仍未盡清楚。本研究利用大花鼠中大腦動脈阻塞的實驗模式，於缺血再灌注傷害後立即局部投予SHH能夠有意義地減少梗塞區域的體積，而預先投予smoothened (Smo)的抑制劑cyclopamine阻斷SHH的作用，則會抑制此現象。本研究結果顯示，局部投予SHH，在1天時有意義增加遭受缺血及再灌注傷害側的Patched1 (Ptc1) 蛋白質的表現。在SHH對於血管新生相關因子的作用，本研究發現SHH的給予不會改變VEGF mRNA和蛋白質表現，但在1天和3天時，分別有意義地增加Ang2 mRNA和蛋白質的表現。而在NOS的mRNA和蛋白質表現量，研究結果顯示給予SHH治療在1天時使nNOS mRNA表現上升，並且有意義地增加nNOS和eNOS的蛋白質表現量。綜合以上的結果，本研究發現，當大花鼠遭受局部中樞缺血及再灌注傷害時，SHH是一個有效的神經保護劑，其神經保護作用可能是藉由其在傷害發生早期增加具保護作用的eNOS蛋白質表現、以及增加nNOS mRNA和蛋白質的表現。此外，Ptc1蛋白質表現量的增加與Ang2 mRNA和蛋白質的表現量有意義的上升，顯示血管新生的作用也可能為SHH在中樞神經保護作用的機轉之一。Sonic hedgehog (SHH) is a highly conserved protein and it plays a key role in regulating vertebrate organogenesis, such as in the growth of digits on limbs and organization of the brain. SHH signaling pathway promotes angiogenisis during the tissuses or organs suffer the ischemia-reperfusion (I/R) injury. Recently investigations show clearly that the SHH protein increased during ischemic injury of kidney and liver. The activation of SHH signaling pathway indirectly upregulated the expression of vascular endothelial growth factor (VEGF), angiopoietin1 and 2 (Ang1 and Ang2), and could reduse the damages from ischemia and reperfusion. Transfered the SHH gene to myocardium after infarction, Kusano’s reserch showed, has cardioprotective effects include the enhancement of angiogenesis, inhibited myocardium fibrosis and prevented the process of apoptosis. Many stusies showed the effects of SHH against the I/R injury. However, the effects of SHH on brain I/R injury is unclear. In this study we used the middle cerebral artery occlusion model to investigate neuroprotective effect of SHH by local application of SHH on the area of focal cerebral ischemia. Results showed that treated with SHH reduced the brain infarction, while pretreated with cyclopamine, the inhibitor of the smoothened (Smo) receptor, blocked the neuroprotective effect of SHH. In addition, local application of SHH increased the Ptc1 protein expression in the first day. SHH increased the Ang2 mRNA expression at the first day and enhanced the Ang2 protein level at the third day without affecting the expression of VEGF mRNA and protein. We also found that SHH increased the mRNA expression of nNOS at the first day and enhanced the nNOS and the eNOS protein levels. In conclusion, SHH is an effective neuroprotective agent. The application of SHH increased the protective eNOS protein and the nNOS protein expression at the beginning of the I/R injury. In addition, the protective mechanisms maybe include the indirectly angiogenesis effect, as the results showed the upregulation of the Ang2 mRNA and protein levels.thesis