抑制人類多瘤性病毒,BK病毒,感染之中藥篩檢
張德卿
國立中正大學
人類多瘤性病毒，BK病毒（BKV），屬於DNA病毒。BKV感染人類通常沒有明顯臨床症狀，並將潛伏於腎臟組織。當寄主免疫狀況低下或被抑制時，病毒會再活化，進而隨著被感染的細胞排放於尿液。腎臟移植病人常使用免疫抑制劑以降低器官排斥反應，卻常使BKV再活化，在腎臟組織大量複製，進而造成溶解性感染，導致腎衰竭，此乃稱為多瘤性病毒相關腎病變（Polyomavirus-associated nephropathy，PVN）。其發生率約為5-10％，卻在這些病人造成大於50％的移植失敗率。截至目前為止，仍缺乏有效抗BKV的藥物。鑒於PVN在臨床上的重要性，我們想尋找有效抑制BKV活性的中藥，以提供治療PVN的新藥物。我們將以抑制BKV VP1類病毒殼體的血液凝集及BKV調控區報告基因活性，篩選出可抑制BKV與細胞受體吸附及轉錄因子結合至BKV調控區的能力的中藥。再以同步定量PCR方法及免疫螢光染色，確認篩選出的中藥抑制BKV感染細胞的能力。同時也以同步定量PCR方法及免疫組織染色，確認中藥抑制BKV感染動物的動物模型試驗。接著依據在細胞培養所確認的中藥，開始分離中藥各部份，再以抑制血液凝集及BKV複製能力，確認各成份活性；再繼續進入細胞培養及動物模型試驗。之後以儀器分析及化學修飾反應鑑定中藥內有效成份及其結構式；同時依不同比率混合各有效成份，再進行上述抑制BKV活性的實驗，以期找到更有效之成份組合，提供臨床上治療因BKV活化而引起的疾病的新藥物。
關鍵字：BK病毒；多瘤性病毒相關腎病變；中藥抗病毒
Screening of Chinese herbal medicine to inhibit human polyomavirus, BK virus, infection
Deching Chang
National Chung Cheng University
Human polyomvirus, BK virus (BKV), is a small DNA virus. After primary infection, BKV remains latent in the kidney. BKV may be reactivated and shed into urine under immunosuppressiong conditions. When immunosuppressive drugs are introduced to renal transplant reciepients to reduce immune rejection, BKV will be reactivated. Reactivation of BK virus with high level viral replication results in lytic disruption of kidney cells, leading to renal dysfunction and loss of graft in these patients. This is so called polyomavirus-associated nephropathy (PVN). There are no effective drugs available for anti-BKV reactivation now. PVN is still an emerging threat to renal transplants. Chinese herbal medicine has been shown to be effective in treatment of virus infection, therefore, the purpose of this study is to search effective drugs for anti-BKV reactivation from Chinese herbal medicine. In vitro screening of herbal extracts on BKV infection inhibition will be evaluated by inhibiting the hemagglutination activity (HAI) of BKV and the BKV replication activity. Inhibition of BKV infection to vero cells will be confirmed by real-time PCR and immunofluorescence assay to determine the BKV viral load and the expression of viral proteins.
關鍵字：BK virus；Polyomavirus-associated nephropathy (PVN)；Herbal medicine antivirus
