Effect of Panax notoginseng Fermented with Lactic Acid Bacteria on the Human Hepatoma Cells in SCID Mice
|Keywords:||三七;乳酸菌;皂苷;抗肝腫瘤;SCID mice;Panax notoginseng;LAB;saponins;anti-hepatoma;SCID mice||Issue Date:||2006||Abstract:||
三七(Panax notoginseng (Burk.) F. H. Chen)乃五加科人參屬，為中國傳統名貴藥材之一；目前已被證實具有鎮痛、護肝、抗發炎、抗血栓、降血壓、保護心肌與抗腫瘤等生理功效，其主要生理活性物質為多醣（一次代謝產物）和皂苷（二次代謝產物）。文獻指出皂苷可透過微生物的代謝提升其生理活性，故本實驗以三七為基質，利用乳酸菌醱酵進行生物性轉化，探討最佳醱酵條件(包括醱酵時間、溫度和pH值)，再藉由體外和體內試驗評估醱酵產品抑制肝癌細胞株生長之功效，並分析皂苷成分變化，期望提升三七在抗肝癌方面之效用。本研究之體外實驗結果發現，以6.6 公升醱酵槽執行醱酵時，在37℃、不控制pH值，醱酵48小時後可得到具最佳抑制肝癌細胞功效之醱酵液（Hep 3B與Hep G2之IC50分別為226 μg/ml、149 μg/ml）。同時，將最具生理活性效果之三七乳酸菌醱酵液以甲醇萃取後，再以LC–ESI-MS/ MS結合CID進行其結構分析與鑑定，可得到protopanaxtriol、ginsenoside Rh1、notoginsenoside R2與ginsenoside Rf四種皂苷；其中protopanaxtriol與ginsenoside Rh1之抗癌效果更已被證實。此部分結果可作為日後純化分離得到更具抑制肝癌細胞增生、降低對正常細胞毒性之產品之參考。後續將以動物模式進一步探討醱酵液抗肝癌之功效。
Panax notoginseng (Burk.) F. H. Chen, is one of the most famous traditional Chinese medicines. It belongs to the Araliaceae family. Its medicinal properties are sweet and slightly bitter in flavor and warm in nature. The main bioactive compound was regarded as saponins, and was used for treatment of diseases of cardiovascular, central nervous, and hematopoietic systems. Cancer prevention and hepatoprotection by saponins have received increasing attention. It has been found that the ginsenosides could be transformed by human intestinal bacteria to metabolites with improvement of anti-carcinogenic activities. Biotransformation of saponins of P. notoginseng by lactic acid bacteria (LAB) is a possible way to enhance its anti-hepatoma activity. The objectives of this study are (1) to establish the optimal conditions, such as fermentation time, temperature and pH value, for the lactic acid fermentation of P. notoginseng, to enhance the anti-hepatoma function of the fermented products; (2) to investigate the bioactive components in the fermentation products which are responsible for the anti-hepatoma activity; (3) to elucidate the inhibitory effect of the fermented products on the growth of hepatoma cells by means of in vitro and in vivo studies.
The present study established the optimal fermentation conditions for the inhibitory effects of fermentative broth on hepatoma cells. The best inhibitory effects were reached after fermentation with LAB for 48 hours at 37℃, and leaving the pH value uncontrolled when carrying out the fermentation in a 6.6 liter bioreactor. The IC50 of Hep 3B and Hep G2 were 226 μg/ml and 149 μg/ml, respectively. Meanwhile, the identification of the saponins in the fermentative broth of P. notoginseng
was elucidated by using LC–ESI-MS/ MS and CID. Based on matching their detection of the saponin molecular weight, and the fragment ions of the molecular ion obtained in the CID experiments with data reported in the literature. Further study will focus on the animal experiments.
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