靈芝，俗稱靈芝草，自古以來在中醫上被認為是能延年益壽、長生不老的上等良藥，「本草綱目」亦記載：「靈芝安心魂，治胸中結，益心氣，人心生血，助心充脈。」，因此數千年來在亞洲地區被人民視為具有神奇功效的「仙草」。靈芝屬於高等真菌，依其顏色或品種可分為六種。靈芝的成分包括多醣體、核苷酸、多肽胺基酸、生物鹼類、微量金屬如鍺以及三萜纇。靈芝已被報導具有抑癌、增強免疫力等功能，對於心血管系統亦多有報導：靈芝具強心作用，並可透過降低膽固醇、血栓、血壓達到擴張血管、增強血流提升循環系統之功效。近來在各項靈芝成分的研究中指出合成的靈芝酸衍生物能透過活化抗氧化機制達到保護心臟細胞的效果。此外，靈芝也被證實可以降低因心肌梗塞造成的心臟傷害，然而關於靈芝的活性有效成分在心臟之保護機轉目前尚不清楚。本篇研究將透過細胞實驗來探究純化之靈芝酸粗萃取物(ganoderic acids, GAs)是否能透過降低細胞內氧化壓力達到避免心臟受傷害之效果。我們利用養分缺乏模式(glucose/ serum deprivation)和過氧化氫(hydrogen peroxide, H2O2)當作氧化劑來增加細胞內活性氧分子(reactive oxygen species, ROS)，並測試靈芝酸是否具有保護效果及其作用。研究結果顯示靈芝酸在H9c2心肌細胞與由仔鼠心臟分離出的初級心臟細胞都能有效地減少因H2O2所引起的細胞凋亡現象。另外，在養分缺乏的情形下心肌細胞內的活性氧分子會增加，而在給予靈芝酸和長時間缺乏養分/生長因子後，靈芝酸具有保護細胞的能力。隨著養分/生長因子缺乏時間增加，細胞內p38蛋白磷酸化、活化態的caspase-3及caspase-9蛋白都會增加，而靈芝酸可以有效降低p38磷酸化與caspase-3和caspase-9之蛋白表現量。此外，靈芝酸也可以透過清除氧化壓力以減少自噬作用(autophagy)的產生。我們進一步利用過氧化氫模擬大量氧化壓力的環境，結果發現給予靈芝酸前處理4小時能顯著的減低細胞凋亡。同時，靈芝酸也能降低因H2O2而升高之p38磷酸化，抑制p38活性以避免心肌細胞受到過氧化氫的傷害。綜合上述結果顯示靈芝酸可透過抑制p38減少細胞凋亡，保護心肌細胞免於不當刺激所造成之死亡，進而使細胞存活。本研究主題之結果能提供未來臨床輔助性治療之契機。

Ganoderma, also known as lingzhi, is one of the most common traditional Chinese herbs. Ganoderma mainly contains polysaccharides, nucleotides, peptides, alkaloid, minor metal such as germanium and triterpenoids responsible for its bioactivities. Accumulating evidence shows that lingzhi can suppress tumor growth and promote immune system and is beneficial to cardiovascular system mainly by improving circulation efficiency through lowering cholesterol, thrombosis, and blood pressure. More recently, the protective antioxidative activities of synthetic triterpenoid derivates were reported in cardiac cells in vitro. Besides, it has been identified that Ganoderma can ameliorate heart damage caused by myocardial infarction via mitochondria and lowering oxdative stress in vivo. However, its molecular mechanism of active components in regulating cardiomyocyte survival is still unclear. We hypothesized that purified lingzhi extract, ganoderic acids (GAs) may prevent hearts from damage through reducing cellular oxidative stress. We challenged cardiomyocytes with glucose/serum starvation or hydrogen peroxide (H2O2) to increase cellular reactive oxygen species (ROS) to test this hypothesis. It has been shown that ROS levels were elevated under serum or glucose starvation in cardiomyocytes. We tested the effect of GAs in cardiomyocytes under long-term deprivation of both serum and glucose, and found that GAs alleviated the apoptosis-induced by lack of nutrients and survival factors. Moreover, cellular phospho-p38 and cleaved caspase-3 protein levels increased in a time-dependent manner after starvation treatment. And GAs effectively decreased the phosphorylation of p38, and the cleavage of caspases-3 and -9 caused by nutrient deficiency. Furthermore, GAs reduced autophagosomes to remain cardiac cells survival. More, cell apoptosis induced by H2O2 was markedly attenuated by GAs pretreatments for 4 hr. GAs also attenuated the p-p38 protein levels induced by H2O2. These results indicated that GAs can protect the cardiac cell from stress-induced apoptosis through inhibiting p38 activation. The study here demonstrates, for the first time, the cardioprotective bioactivities and their clinical implication of ganoderic acids derived from Ganoderma tsugae.

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