丹參酮 IIA 對感染幽門螺旋桿菌胃腺癌細胞凋亡之機轉

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丹參酮 IIA 對感染幽門螺旋桿菌胃腺癌細胞凋亡之機轉

Mechanisms of tanshinone IIA in the Helicobacter pylori-infected MKN45 cell apoptosis

許喻捷 , Masters　　Advisor：王苑春　　

繁體中文

丹參酮 IIA ；幽門螺旋桿菌；胃腺癌細胞；細胞凋亡； Tanshinone IIA ； Helicobacter pylori ； gastric adenocarcinoma cell ； apoptosis

Abstract │ Reference (186) │ Altmetrics

Abstract 〈TOP〉

幽門螺旋桿菌 (Helicobacter pylori) 與慢性胃炎、胃潰瘍及胃癌具高度關聯性，慢性胃炎為胃癌發展過程之關鍵，一般認為細胞凋亡則為胃腺癌之重要抑制路徑，1994 年聯合國世界衛生組織 (WHO)將幽門螺旋桿菌列為第一級致癌因子。丹參酮 IIA (tanshinone IIA) 為丹參的脂溶性二萜類成分，具有抗發炎及抗癌等生理活性。在我們先前研究中，丹參酮 IIA 能有效的抑制幽門螺旋桿菌感染造成宿主細胞發炎，因此本研究接續先前的研究，探討丹參酮 IIA 對被幽門螺旋桿菌感染之 MKN45 胃腺癌細胞於細胞凋亡之影響。

本研究結果顯示，以 16 μg/mL 丹參酮 IIA 處理，可顯著的降低MAPK (Mitogen-activated protein kinases) 路徑之 JNK1/2 及 p38 蛋白質的表現，亦能抑制 5-脂氧合酶 (5-Lipoxygenase) 蛋白質的表現。上述三種蛋白質的表現分別為控制組 (0 μg/mL) 的 70、67 及 65%。以 16 μg/mL 丹參酮 IIA 處理 可顯著的增加細胞週期分佈之 subG0/G1，比例，為控制組之 246%；以 16 μg/mL 丹參酮 IIA 處理，可顯著的促進粒 線 體 膜電 位 下降 ， 為 控制 組的 71%， 並增加 Bax、 細 胞質cytochrome c 及 caspase-9 等蛋白質的表現 分別為控制組的 292 200，、及 173%，以及降低 Bcl-2、粒線體之 cytochrome c 及 pro-caspase-3 等蛋白質的表現，分別為控制組的 55、56 及 36%。

丹參酮 IIA 能有效抑制 MAPK 路徑之相關蛋白質表現，進而抑制下游發炎因子，致使感染幽門螺旋桿菌之宿主細胞發炎受抑制。宿主細胞的發炎因子受抑制時會活化促凋亡蛋白 Bax，丹參酮 IIA 能有效抑制抗凋亡蛋白 Bcl-2 及促進促凋亡蛋白 Bax 等表現，並促進粒線體膜電位下降，致使粒線體之 cytochrome c 釋放至細胞質，亦能有效的活化下游 pro-caspase-3 及 caspase-9 蛋白質表現。綜合本研究結果，丹參酮 IIA 能有效的抑制感染 H. pylori 宿主細胞發炎之訊號傳導，同時亦有效地促進感染 H. pylori 宿主細胞之凋亡；丹參酮 IIA 具有阻斷感染 H. pylori 宿主細胞演化成為胃腺癌細胞之高度潛力。

Parallel Abstract 〈TOP〉

Helicobacter pylori is strongly associated with chronic gastritis, peptic ulcer, and gastric cancer. Chronic gastritis plays an important role in the progression of gastric cancer. Apoptosis is a key factor to inhibit gastric cancer progression. World Health Organization (WHO) defined H. pylori as a group I carcinogen in 1994. Tanshinone IIA, an diterpene quinones, was isolated from the roots of Salvia miltiorrhiza, having anti-inflammatory and anti-cancer activities. In our previous study, tanshinone IIA exhibited anti-inflammatory effect in the H. pylori-infected MKN45 cell. Therefore, in this study, the effects of tanshinone IIA on the H. pylori-infected gastric adenocarcinoma MKN45 cell apoptosis were investigated.

Tanshinone IIA treatment (16 μg/mL) significantly inhibited MAPK pathway (p38 and JNK1/2) and 5-LOX protein expressions; 70, 67, and 65% of that of the controls (0 μg/mL treatment), respectively. Tanshinone IIA treatment (16 μg/mL) significantly increased subG0/G1 phase of the cell cycle, 246% of that of the control. The mitochondrial membrane potential significantly collapsed, 71% of that of the control at 16 μg/mL of treatment dose. Tanshinone IIA (16 μg/mL) significantly increased Bax, cytosolic cytochrome c, and caspase-9 protein expressions; 292, 200 and 173% of that of controls, respectively. Tanshinone IIA (16 μg/mL) significantly decreased Bcl-2, mitochondrial cytochrome c, and pro-caspase-3 protein expressions; 55, 56, and 36% of that of controls, respectively.
   
Tanshinone IIA significantly inhibited MAPK pathway’s protein expressions resulted in the inhibition of downstream inflammatory factors, and then H. pylori-infected cell inflammation was significantly inhibited. Inflammatory factor inhibition was responsible for the activation of pro-apoptotic protein (Bax). Tanshinone IIA significantly inhibited Bcl-2 and increased Bax protein expressions, and then induced mitochondrial membrane potential collapsed. Subsequently, cytochrome c was released from mitochondria to cytosol resulted in the activation of caspase-9 and pro-caspase-3, and then triggered MKN45 cell apoptosis.
  
In conclusion, tanshinone IIA significantly inhibited inflammatory signaling transduction and induced apoptosis in the H. pylori-infected MKN45 cells. Tanshinone IIA showed great potential to prevent gastric cancer progression in the H. pylori-infected cells.

Reference ( 186 ) 〈TOP〉

陳冠宇。丹參酮 IIA 對幽門螺旋桿菌感染胃腺癌細胞及巨噬細胞發炎之抑制。國立中興大學食品暨應用生物科技研究所碩士學位論文。2013。
連結：
Adams, J. M., & Cory, S. The Bcl-2 protein family: arbiters of cell survival. Science. 1998a; 281, 1322-1326.
連結：
Ahmed, A., Smoot, D., Littleton, G., Tackey, R., Walters, C. S., Kashanchi, F., Allen, C. R., & Ashktorab, H. Helicobacter pylori inhibits gastric cell cycle progression. Microbes Infect. 2000; 2, 1159-1169.
連結：
Ali, M., Khan, A. A., Tiwari, S. K., Ahmed, N., Rao, L. V., & Habibullah, C. M. Association between cag-pathogenicity island in Helicobacter pylori isolates from peptic ulcer, gastric carcinoma, and non-ulcer dyspepsia subjects with histological changes. World J Gastroenterol. 2005; 11, 6815-6822.
連結：
Allison, C. C., Kufer, T. A., Kremmer, E., Kaparakis, M., & Ferrero, R. L. Helicobacter pylori induces MAPK phosphorylation and AP-1 activation via a NOD1-dependent mechanism. J Immunol. 2009; 183, 8099-8109.
連結：

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