動脈粥狀硬化(atherosclerosis)是一個有很多因素且複雜的致病過程，氧化和發炎這兩個因素對於動脈硬化發展過程扮演重要的角色，氧化壓力的增加對於內皮細胞失去功能和動脈粥狀硬化有很大的關係。前胸腺素(prothymosin a)是一個廣泛分布的小分子核蛋白，早期的研究主要在於其參與細胞增生及免疫調控。最近的研究指出前胸腺素可能參與調控一些保護身體對抗氧化的基因，例如第一型血基質氧化酶(HO-1)。 第一型血基質氧化酶是鐵代謝的速率決定步驟，將鐵代謝成一氧化碳、 膽綠素(biliverdin)、亞鐵離子。很多研究指出第一型血基質氧化酶和其產物可以對抗氧化壓力。我們推測前胸腺素的過量表現可以引起抗氧化基因的表現，而去抑制ApoE基因缺陷老鼠自發性形成的粥狀動脈硬化過程。在我們的研究，我們使用攜帶前胸腺素基因的腺病毒(命名為Ad/ProT)，嘗試去治療ApoE基因缺陷老鼠自發性形成的粥狀動脈硬化。在細胞實驗中，我們觀察到過量表現前胸腺素的確可以促使抗氧化基因產生，例如第一型血基質氧化酶或是超氧化物歧化酶(SOD)，並且減少過氧化氫引起的氧化傷害。再者，前胸腺素可以增加內皮型一氧化氮合成脢(eNOS)，並且增加一氧化氮的產生。在動物實驗中，我們將攜帶前胸腺素的腺病毒直接打入麻醉的小鼠心臟中。四個禮拜後，我們犧牲老鼠，分析動脈硬化的程度，我們發現過量表現前胸腺素可以抑制動脈硬化的形成。

Atherosclerosis is a multifactorial and complex pathological process. Oxidation and inflammation are two crucial events which are involved in the development of atherosclerotic lesions. Increased oxidative stress plays a key role in the development of endothelial dysfunction and atherosclerosis. Prothymosin a (ProT) is an abundantly expressed small nuclear protein. Early studies found that it is involved in proliferation of mammalian cells and has immunomodulatory functions. Recently, ProT was shown to be involved in regulating expression of the oxidative stress-protective genes, such as heme oxygenase-1 (HO-1). HO is the rate-limiting enzyme in heme breakdown to generate carbon monoxide, biliverdin, and free ferrous iron. Accumulating evidence has shown that inducible HO (HO-1) and its products function as adaptive molecules against oxidative insults. We hypothesized ProT overexpression might induce antioxidative genes and reduce atherosclerosis progression in apolipoprotein E-deficient mice. In our study, an adenoviral vector expressing ProT, designated “Ad/ProT”, was generated for gene therapy in ApoE-deficient mice. In vitro study, overexpression of ProT enhanced some antioxidative genes, such as HO-1 and SOD, and reduced H2O2-induced oxidative damage. Furthermore, ProT enhanced eNOS expression and increased NO release. In vivo, Ad/ProT was transfered in arteries by direct injection into the left ventricles of ApoE-deficient mice. After 4 weeks, we analysed lesion formation in the aortic sinus. We observed overexpression of ProT could reduce lesion process compared with the Ad/LacZ group.

Cid, M. C., Schnaper, H. W., and Kleinman, H. K. (2002). Estrogens and the vascular endothelium. Ann N Y Acad Sci 966, 143-157.
Cordero, O. J., Sarandeses, C. S., Lopez, J. L., Cancio, E., Regueiro, B. J., and Nogueira, M. (1991). Prothymosin alpha enhances interleukin 2 receptor expression in normal human T-lymphocytes. Int J Immunopharmacol 13, 1059-1065.
Cotter, M. A., 2nd, and Robertson, E. S. (2000). Modulation of histone acetyltransferase activity through interaction of epstein-barr nuclear antigen 3C with prothymosin alpha. Mol Cell Biol 20, 5722-5735.
Cushing, S. D., Berliner, J. A., Valente, A. J., Territo, M. C., Navab, M., Parhami, F., Gerrity, R., Schwartz, C. J., and Fogelman, A. M. (1990). Minimally modified low density lipoprotein induces monocyte chemotactic protein 1 in human endothelial cells and smooth muscle cells. Proc Natl Acad Sci U S A 87, 5134-5138.
Dhakshinamoorthy, S., and Jaiswal, A. K. (2001). Functional characterization and role of INrf2 in antioxidant response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene. Oncogene 20, 3906-3917.
Dimmeler, S., Rippmann, V., Weiland, U., Haendeler, J., and Zeiher, A. M. (1997). Angiotensin II induces apoptosis of human endothelial cells. Protective effect of nitric oxide. Circ Res 81, 970-976.
Evstaf'eva, A. G., Karapetian, R. N., Rubtsov Iu, P., Filonov, G. S., Abaeva, I. S., Fateeva, T. V., Mel'nikov, S. V., Chichkova, N. V., and Vartapetian, A. B. (2005). [Novel functions of the well-known protein--prothymosin alpha is involved in protection of cells against apoptosis and oxidative stress]. Mol Biol (Mosk) 39, 729-745.
Faraci, F. M., and Didion, S. P. (2004). Vascular protection: superoxide dismutase isoforms in the vessel wall. Arterioscler Thromb Vasc Biol 24, 1367-1373.
Freire, J., Covelo, G., Sarandeses, C., Diaz-Jullien, C., and Freire, M. (2001). Identification of nuclear-import and cell-cycle regulatory proteins that bind to prothymosin alpha. Biochem Cell Biol 79, 123-131.
Fukai, T., Folz, R. J., Landmesser, U., and Harrison, D. G. (2002). Extracellular superoxide dismutase and cardiovascular disease. Cardiovasc Res 55, 239-249.
Gast, K., Damaschun, H., Eckert, K., Schulze-Forster, K., Maurer, H. R., Muller-Frohne, M., Zirwer, D., Czarnecki, J., and Damaschun, G. (1995). Prothymosin alpha: a biologically active protein with random coil conformation. Biochemistry 34, 13211-13218.
Gryglewski, R. J., Palmer, R. M., and Moncada, S. (1986). Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor. Nature 320, 454-456.
Gupte, S. A., Kaminski, P. M., Floyd, B., Agarwal, R., Ali, N., Ahmad, M., Edwards, J., and Wolin, M. S. (2005). Cytosolic NADPH may regulate differences in basal Nox oxidase-derived superoxide generation in bovine coronary and pulmonary arteries. Am J Physiol Heart Circ Physiol 288, H13-21.
Hamilton, C. A., Miller, W. H., Al-Benna, S., Brosnan, M. J., Drummond, R. D., McBride, M. W., and Dominiczak, A. F. (2004). Strategies to reduce oxidative stress in cardiovascular disease. Clin Sci (Lond) 106, 219-234.
Hirayama, A., Yoh, K., Nagase, S., Ueda, A., Itoh, K., Morito, N., Hirayama, K., Takahashi, S., Yamamoto, M., and Koyama, A. (2003). EPR imaging of reducing activity in Nrf2 transcriptional factor-deficient mice. Free Radic Biol Med 34, 1236-1242.
Hisamoto, K., Ohmichi, M., Kurachi, H., Hayakawa, J., Kanda, Y., Nishio, Y., Adachi, K., Tasaka, K., Miyoshi, E., Fujiwara, N., et al. (2001). Estrogen induces the Akt-dependent activation of endothelial nitric-oxide synthase in vascular endothelial cells. J Biol Chem 276, 3459-3467.
Hodgin, J. B., Krege, J. H., Reddick, R. L., Korach, K. S., Smithies, O., and Maeda, N. (2001). Estrogen receptor alpha is a major mediator of 17beta-estradiol's atheroprotective effects on lesion size in Apoe-/- mice. J Clin Invest 107, 333-340.
Immenschuh, S., and Schroder, H. (2006). Heme oxygenase-1 and cardiovascular disease. Histol Histopathol 21, 679-685.
Itoh, K., Wakabayashi, N., Katoh, Y., Ishii, T., Igarashi, K., Engel, J. D., and Yamamoto, M. (1999). Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev 13, 76-86.
Jaiswal, A. K. (2004). Nrf2 signaling in coordinated activation of antioxidant gene expression. Free Radic Biol Med 36, 1199-1207.
Juan, S. H., Lee, T. S., Tseng, K. W., Liou, J. Y., Shyue, S. K., Wu, K. K., and Chau, L. Y. (2001). Adenovirus-mediated heme oxygenase-1 gene transfer inhibits the development of atherosclerosis in apolipoprotein E-deficient mice. Circulation 104, 1519-1525.
Karapetian, R. N., Evstafieva, A. G., Abaeva, I. S., Chichkova, N. V., Filonov, G. S., Rubtsov, Y. P., Sukhacheva, E. A., Melnikov, S. V., Schneider, U., Wanker, E. E., and Vartapetian, A. B. (2005). Nuclear oncoprotein prothymosin alpha is a partner of Keap1: implications for expression of oxidative stress-protecting genes. Mol Cell Biol 25, 1089-1099.
Kobayashi, A., Ohta, T., and Yamamoto, M. (2004). Unique function of the Nrf2-Keap1 pathway in the inducible expression of antioxidant and detoxifying enzymes. Methods Enzymol 378, 273-286.
Lin, S. J., Shyue, S. K., Hung, Y. Y., Chen, Y. H., Ku, H. H., Chen, J. W., Tam, K. B., and Chen, Y. L. (2005). Superoxide dismutase inhibits the expression of vascular cell adhesion molecule-1 and intracellular cell adhesion molecule-1 induced by tumor necrosis factor-alpha in human endothelial cells through the JNK/p38 pathways. Arterioscler Thromb Vasc Biol 25, 334-340.
Madamanchi, N. R., Hakim, Z. S., and Runge, M. S. (2005a). Oxidative stress in atherogenesis and arterial thrombosis: the disconnect between cellular studies and clinical outcomes. J Thromb Haemost 3, 254-267.
Madamanchi, N. R., Vendrov, A., and Runge, M. S. (2005b). Oxidative stress and vascular disease. Arterioscler Thromb Vasc Biol 25, 29-38.
Malicet, C., Giroux, V., Vasseur, S., Dagorn, J. C., Neira, J. L., and Iovanna, J. L. (2006). Regulation of apoptosis by the p8/prothymosin alpha complex. Proc Natl Acad Sci U S A 103, 2671-2676.
Martini, P. G., Delage-Mourroux, R., Kraichely, D. M., and Katzenellenbogen, B. S. (2000). Prothymosin alpha selectively enhances estrogen receptor transcriptional activity by interacting with a repressor of estrogen receptor activity. Mol Cell Biol 20, 6224-6232.
Martini, P. G., and Katzenellenbogen, B. S. (2003). Modulation of estrogen receptor activity by selective coregulators. J Steroid Biochem Mol Biol 85, 117-122.
McEver, R. P. (1992). Leukocyte-endothelial cell interactions. Curr Opin Cell Biol 4, 840-849.
Miller, F. J., Jr., Gutterman, D. D., Rios, C. D., Heistad, D. D., and Davidson, B. L. (1998). Superoxide production in vascular smooth muscle contributes to oxidative stress and impaired relaxation in atherosclerosis. Circ Res 82, 1298-1305.
Minuz, P., Fava, C., and Cominacini, L. (2006). Oxidative stress, antioxidants, and vascular damage. Br J Clin Pharmacol 61, 774-777.
Morita, T. (2005). Heme oxygenase and atherosclerosis. Arterioscler Thromb Vasc Biol 25, 1786-1795.
Mugge, A., Elwell, J. H., Peterson, T. E., and Harrison, D. G. (1991). Release of intact endothelium-derived relaxing factor depends on endothelial superoxide dismutase activity. Am J Physiol 260, C219-225.
Navab, M., Berliner, J. A., Watson, A. D., Hama, S. Y., Territo, M. C., Lusis, A. J., Shih, D. M., Van Lenten, B. J., Frank, J. S., Demer, L. L., et al. (1996). The Yin and Yang of oxidation in the development of the fatty streak. A review based on the 1994 George Lyman Duff Memorial Lecture. Arterioscler Thromb Vasc Biol 16, 831-842.
Ozono, R. (2006). New biotechnological methods to reduce oxidative stress in the cardiovascular system: focusing on the Bach1/heme oxygenase-1 pathway. Curr Pharm Biotechnol 7, 87-93.
Rajavashisth, T. B., Andalibi, A., Territo, M. C., Berliner, J. A., Navab, M., Fogelman, A. M., and Lusis, A. J. (1990). Induction of endothelial cell expression of granulocyte and macrophage colony-stimulating factors by modified low-density lipoproteins. Nature 344, 254-257.
Ross, R. (1993). The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 362, 801-809.
Segade, F., and Gomez-Marquez, J. (1999). Prothymosin alpha. Int J Biochem Cell Biol 31, 1243-1248.
Stocker, R., and Perrella, M. A. (2006). Heme oxygenase-1: a novel drug target for atherosclerotic diseases? Circulation 114, 2178-2189.
Sumi, D., and Ignarro, L. J. (2003). Estrogen-related receptor alpha 1 up-regulates endothelial nitric oxide synthase expression. Proc Natl Acad Sci U S A 100, 14451-14456.
Thiemermann, C., and Vane, J. (1990). Inhibition of nitric oxide synthesis reduces the hypotension induced by bacterial lipopolysaccharides in the rat in vivo. Eur J Pharmacol 182, 591-595.
Tousoulis, D., Antoniades, C., Koumallos, N., Marinou, K., Stefanadi, E., Latsios, G., and Stefanadis, C. (2006). Novel therapies targeting vascular endothelium. Endothelium 13, 411-421.
Tousoulis, D., Antoniades, C., Tentolouris, C., Goumas, G., Stefanadis, C., and Toutouzas, P. (2002). L-arginine in cardiovascular disease: dream or reality? Vasc Med 7, 203-211.
Vora, D. K., Fang, Z. T., Liva, S. M., Tyner, T. R., Parhami, F., Watson, A. D., Drake, T. A., Territo, M. C., and Berliner, J. A. (1997). Induction of P-selectin by oxidized lipoproteins. Separate effects on synthesis and surface expression. Circ Res 80, 810-818.
Wang, H. H., Hung, T. M., Wei, J., and Chiang, A. N. (2004). Fish oil increases antioxidant enzyme activities in macrophages and reduces atherosclerotic lesions in apoE-knockout mice. Cardiovasc Res 61, 169-176.
Xu, G. L., Qian, Z. Y., Yu, S. Q., Gong, Z. N., and Shen, X. C. (2006). Evidence of crocin against endothelial injury induced by hydrogen peroxide in vitro. J Asian Nat Prod Res 8, 79-85.
Yang, C. H., Murti, A., Baker, S. J., Frangou-Lazaridis, M., Vartapetian, A. B., Murti, K. G., and Pfeffer, L. M. (2004). Interferon induces the interaction of prothymosin-alpha with STAT3 and results in the nuclear translocation of the complex. Exp Cell Res 298, 197-206.
Yokoyama, M. (2004). Oxidant stress and atherosclerosis. Curr Opin Pharmacol 4, 110-115.