以初代培養鴨肝細胞篩選抗親肝DNA病毒藥物之可行性研究

A Study on the Possibility of Screening the Drug with Antihepadnaviral Effect by Primary Duck Hepatocyte Culture

沈永紹(Yong-Shau Shien) ； 周世認(Shyh-Renn Chou) ； 黃冠棠(Guan-Tarn Huang)

《中華民國獸醫學會雜誌》 22卷3期 (1996/06) Pp. 192-204

https://doi.org/10.7009/JTCSVS.199606.0192

摘要

本研究用由4種具有強力抗鴨B型肝炎病毒聚合酶（anti-duck hepatitis B virus endogenous DNA polymerase, anti-DHBV-DNAP）效果之中藥分離之11種抽出物，檢討初代培養之先天性感染DHBV雛鴨肝細胞系統做為篩選具抗親肝DNA病毒效力藥物之可行性。經觀察細胞形態及測定培養液中乳酸鹽去氫酶（lactate dehydrogenase）及氨基丙酸轉氨酶（alanine aminotransferase）活性值變化並測定肝細胞內及培養液中DHBV-DNA含量以及測定細胞中DHBV複製中間物之消長。試驗結果顯示：初代培養之先天感染DHBV雛鴨肝細胞做為篩選抗親肝DNA病毒藥物有下述優點：肝細胞分離後不必再經人工感染DHBV；可評估藥物是否具有細胞毒性；可知藥物是否可通過細胞膜；可推測藥物經細胞代謝後之藥效變化；及可推測藥物抑制DHBV複製之作用機轉。由上述結果言，本篩選系統似可成為有大量藥物要篩選時之一種經濟、省時、半試管內性及半活體性之篩選方法。

關鍵字

鴨B型肝炎病毒 ； 鴨肝細胞培養 ； 抗親肝DNA病毒藥物

並列摘要

The aim of this study was evaluate the possibility of primary culture hepatocytes separated from congenitally infected with duck hepatitis B virus (DHBV) ducklings as a tool for screening the drug with anti-hepadnaviral activity. Eleven extracts from four medicinal herbs with strong anti-DHBV endogenous DNA polymerase(DNAP) activity were employed in this study. Changes of the following items were carefully investigated: the hepatocyte morphology; the activities of lactate dehydrogenase and alanine aminotransferase in the culture media; the DHBV-DNA concentration in hepatocytes and in the culture media; and the intermediate products of DHBV-DNA in hepatocytes. Result revealed that this screening system had following excellent characters: not necessary to infect the cells after separation; easy to evaluate the toxicity and the ability to cross the cell membrane of the drugs; and easy to infer the mechanism of anti-DHBV replication of the drug. Therefore, this screening system could be an economic, time-saving, Semi-in vitro and semi-in vivo method to screen a huge number of drugs.

並列關鍵字

DHBV ； Duck hepatocyte culture ； Anti-hepadnaviral drugs