“Exchange protein activated by cAMP (Epac1)”, 是鳥嘌呤交換蛋白之一種, 是cAMP activated for small RapGTPases, 相關功能反應已知如下，細胞附著力，胰島素分泌，調節血管內皮細胞的通透性，以及其他反應 (Nature 1998; Science 1998; J Physiol 2006). 我們實驗室發現運動會增加Epac1但 Epac1在運動的角色仍是未知的。(1/5) 實驗方法: 第一年探討運動治療對糖尿病心臟與腦部的Epa1 和Epac1下游Rap1 GTPase-activating protein途徑, 與其血管增生及抗凋亡途徑探討包含三組動物模式 [G1: non DM;G2: STZ-induced DM; G3: STZ-induced DM C57BL/6JNarl mice+ Swimming exercise 3 month (n=30 each)]. (2/4) 實驗方法: 第二年探討Lentiviral Epac1 gene transfer (LV-Epac1)是否藉由MMP2 and MMP9調節並誘發骨髓幹細胞自誘導機轉對糖尿病心臟與腦部的產生血管增生及抗凋亡的機轉，實驗可分七組 [G1: non DM;G2: STZ-induced DM; G3: STZ-induced DM C57BL/6JNarl mice+ Lentiviral Epac1 gene transfer (LV-Epac1);G4: STZ-induced DM MMP-9 knockout mice;G5: STZ-induced DM MMP-9 knockout mice+ Lentiviral Epac1 gene transfer (LV-Epac1);G6: STZ-induced DM MMP-2 knockout mice;G7: STZ-induced DM MMP-9 knockout mice+ Lentiviral Epac1 gene transfer (LV-Epac1) (n=30 each)]. (3/4) 實驗方法: 第三年探討運動治療對螢光骨髓幹細胞自誘導在糖尿病心臟與腦部的修復位置是否與由運動誘發Epac1的位置相同，實驗可分三組配合有無Epac1抑制劑 [G1: non DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G2: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G3: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice+ Swimming exercise 3 month (n=40 each)]. (4/4) 實驗方法: 第四年探討運動治療對螢光骨髓幹細胞自誘導與Epac1在糖尿病心臟與腦部的修復是否藉MMP來調節，，實驗可分五組[G1: non DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G2: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice;G3: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month; G4: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month (n=20) +MMP inhibitor; G5: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month +MMP inhibitor ] 此研究可探討”Exchange protein activated by cAMP (Epac1)” 在運動治療對糖尿病心臟與腦部的幹細胞自誘導機轉與抗凋亡機轉，也提供人類修復心臟與腦部重要機轉與治療的新窗口。
Exchange protein activated by cAMP (Epac1), one of guanine exchange proteins, are cAMP activated for small RapGTPases, and has been associated with responses such as cell adhesion, insulin secretion, regulation of endothelial cell permeability, and other responses (Nature 1998; Science 1998; J Physiol 2006). Epac1 was found upregulated by our lab but effects of Epac1 on exercise training is unknown. (1/4) Method: Effects of Exercise on exchange protein activated by cAMP (Epac1), downstream Rap1 GTPase-activating protein (Rap1-GTP) and anti-apoptosis in diabetic heart and brain will be studied in three group [G1: non DM;G2: STZ-induced DM; G3: STZ-induced DM C57BL/6JNarl mice+ Swimming exercise 3 month (n=30 each)]. (2/4) Method: Lentiviral Epac1 gene transfer (LV-Epac1) induced BMSCs trafficking and anti-apoptosis to diabetic heart and brain via upregulation of MMP2 and MMP9 will be studied in seven groups [G1: non DM;G2: STZ-induced DM; G3: STZ-induced DM C57BL/6JNarl mice+ Lentiviral Epac1 gene transfer (LV-Epac1);G4: STZ-induced DM MMP-9 knockout mice;G5: STZ-induced DM MMP-9 knockout mice+ Lentiviral Epac1 gene transfer (LV-Epac1);G6: STZ-induced DM MMP-2 knockout mice;G7: STZ-induced DM MMP-9 knockout mice+ Lentiviral Epac1 gene transfer (LV-Epac1) (n=30 each)]. (3/4) Method: Colocalization of Exercise-induced GFP-mice BMSCs trafficking and Exercise-induced Exchange protein activated by cAMP (Epac1) in Green Fluorescent Protein (GFP) -bone marrow transplanted mice will be study from three group [G1: non DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G2: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G3: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice+ Swimming exercise 3 month (n=40 each)]. (4/4) Method: Exercise-induced BMSCs trafficking and exchange protein activated by cAMP (Epac1) and downstream Rap1 GTPase-activating protein (Rap1-GTP) in Green Fluorescent Protein (GFP) -bone marrow transplanted mice were blocked by MMP inhibitor will be studied from five group [G1: non DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow mice; G2: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice;G3: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month; G4: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month (n=20) +MMP inhibitor; G5: STZ-induced DM C57BL/6JNarl Green Fluorescent Protein (GFP) -bone marrow transplanted mice+ Swimming exercise 3 month +MMP inhibitor ]. This study will provide a novel and therapeutic mechanism of Epac1 in exercise therapy to repair diabetic heart and brain.
