Identification of structures and effects of destruxin on hepatitis B surface antigen production in human hepatoma cells

• Main Authors: Pan, Wei, 潘瑋
• Other Authors: Yeh, Sheau-Farn
• Format: Others
• Language: zh-TW
• Published: 1995
• Online Access: http://ndltd.ncl.edu.tw/handle/63892153106194357994

碩士 === 國立陽明大學 === 生物化學研究所 === 83 === Destruxins是一種環縮月太，本研究分離真菌Metarhiziumanisopliae萃取液，經由高效能液相層析儀純化，並利用質譜儀以及核磁共振技術等，定出兩種Destruxin類似物DestruxinE chlorohydrin和E2 chlorohydrin的結構，其化學式分別為C29H48CIN5O8以及C28H46CIN5O8，構造式分別是cyclo(2,4-dihydroxy-5-chloropentanoic acid -Pro-Ile-(Me-Val)-(Me-Ala)-β-Ala)depsipeptides和cyclo(2,4-dihydroxy-5-chloropentanoic acid-Pro-Val-(Me-Val)-(Me-Ala)-β-Ala)depsipeptides。其中後者尚未在此真菌培養液中被發現過，是一個新的天然物。人類肝癌細胞株經這兩種Destruxin類似物處理48小時後，乙型肝炎病毒表面抗原的產生會受到抑制，其抑制值IC50分別是0.15μM和6.5μM。另外本研究也比較M.anisopliae培養液中其它Destruxin結構類似物的抑制情形，顯示其效果與分子環的大小有關。 Destruxins也是vacuolar-type ATPase的抑制劑，人類肝癌細胞株經Destruxins處理後也可以觀察到胞器酸化的情形被抑，但5分鐘後即恢復原來的酸鹼值，即使再加入Destruxins，這個現象也不再重現。細胞處理Destruxins 40分鐘後，表面抗原會累積在細胞體內，但是經48小時的處理細胞體內的表面抗原並沒有累積的現象，顯示Destruxins可透過抑制vacuolar-type ATPase的活性而暫時延緩表面抗原的輸送。 這些結果顯示，Destruxins抑制乙型肝炎病毒表面抗原的活性可能由於其暫時抑制胞器酸化的情形所直接導致，也可能是透過其它途徑。 Destruxins are a series of cyclodepsipeptides. In this study, two destruxins, Destruxin E chlorohydrin and Destruxin E2 chlorohydrin, the later is a new compound were identified in this study. They were separated from the culture broth of Fungus, Metarhizium anisopliae and purified by HPLC. The structures were characterized by Mass spectroscopy and NMR techniques. he chemical formulas are C29H48CIN5O8 and C28H46CIN5O8, and the structural formulas are cyclo (2, 4-dihydroxy-5-chloropentanoic acid-Pro-Ile-(Me-Val)-(Me-Ala)-β-Ala) depsipeptides and cyclo (2, 4-dihydroxy-5-chloropentanoic acid-Pro-Val-(Me-Val)-(Me-Ala)-β-Ala) depsipeptides. Both of the compounds can suppress the Hepatitis B virus surface antigen (HBsAg) production on Hep 3B cells after 48 hours treatment. The value of inhibition of of HBsAg production with IC50 of Destruxin E chlorohydrin and Destruxin E2 chlorohydrin is 0.15 μM and 6.5 μM, respectively. The suppression of HBsAg production for other known Destruxins isolated from M. anisopliae culture broth has the same suppressive effect. The effect was affected by the size of the 19-membered ring. Destruxins inhibited the activity of vacuolar-type ATPase. The acidification of cell organelles was blocked while treating with Destruxins but recovering after 5 minutes in Hep 3B cell system. The HBsAg accumulated in the cells by treatment with Destruxinsfor 40 minutes, but it did not accumulate for 48 hours-treatment This indicates that destruxins can delay the secretion ofHBsAg by inhibiting the activity of vacuolar-type ATPase. The inhibitory effects of destruxins HBsAg production may be caused by the inhibition of acidification of cell organelles directly It may involve in some other ways.