| Abstract: | Caffeic acid phenethyl ester (CAPE) 是蜂膠中主要成分之ㄧ。而過去研究證實蜂膠具有抗發炎、抗菌及抗病毒、止血的功能、以及抗腫瘤增生等作用。先前本實驗已證實CAPE於較高劑量(>50μM)時可以透過MAPKs Pathway進一步活化p53，引發cytochrome C的釋放及活化caspase-9,caspase-3訊息路徑最後導致神經膠質細胞 (C6-Glioma Cell) 走向細胞凋亡。另外在CAPE較低劑量(<10μM)時，發現可增加C6神經膠質瘤細胞分化蛋白如: GFAP, S-100β的表現。進而抑制細胞惡化的能力。因此CAPE在較高劑量及較低劑量誘導C6-Glioma細胞的凋亡及抑制惡化的訊號路徑機制尚未十分清楚。本次研究利用Western Blot、 HPTLC、DAPI染色、DNA 電泳觀察、螢光光譜儀、Boyden Chamber及 Wound Healing觀察，結果發現在CAPE較高劑量可快速的降低GSH而導致N-SMase活化ceramide表現，進而活化p38 Kinase另外也活化ERK Kinase而促進NGF/p75表現及隨後的JNK Kinase，最後使細胞走向凋亡。另外於低劑量處理細胞後，可促進NGF/p75表現，進而增加GFAP, S-100β,RhoB蛋白表現及抑制MMP-9,2活性而降低轉移惡化的能力。綜合以上結果證實高劑量CAPE可以誘導活化N-SMase/ceramide、MAPK Pathway及NGF/p75最後使細胞走向凋亡，而在低劑量CAPE則可以誘導NGF/p75持續表現，進而增加GFAP, S-100β,RhoB分化蛋白表現及抑制MMP-9,MMP2活性而使細胞轉移侵犯被抑制。
Caffeic acid phenethyl ester (CAPE) which is one of component of propolis, has been reported to possess anti-inflammatory, anti-bacterial, anti-virus, hemostasis and anti-tumor activity. Previous, Our laboratory demonstrated the role of MAPKs signal pathway, p53, cytochrome c released and activated caspase-9,caspase-3 in regulating the CAPE(>50μM)-induced apoptosis in C6-glioma cells. In addition, it demonstrated GFAP, S-100β protein expression mediated blocking cell deterioration by CAPE(<10μM). However, the signals mediating C6-Glioma apoptosis and blocking deterioration by CAPE(>50μM, <10μM) have not yet been clarified. In the present study, by western blot, HPTLC, DAPI Stain, DNA eletrophoresis, fluorescence spectrograph, Boyden chamber and wound healing assay. We concluded result that CAPE (>50μM) induced apoptosis of C6-glioma cell through reduction of GSH, activation of SMase/ceramide, activation of p38 Kinase and upregulation of NGF/p75 cascade, JNK Kinase involving ERK activation . Another , CAPE (<10μM) blocked deterioration of C6 glioma cell through activation of NGF/p75, expression of well differentiated protein (GFAP, S-100β, RhoB) and reduction of (MMP-2,MMP-9) activity. In conclusion, the role of N-SMase/ceramide, MAPKs signal pathway and NGF/p75 in regulating the CAPE (>50μM) induced apoptosis of C6-glioma cells. In addition, the role of NGF/p75, differentiated protein (GFAP, S-100β, RhoB) and (MMP-2,MMP-9) in regulating the CAPE (<10μM) blocking deterioration of C6-glioma cells. |
