細胞凋亡(apoptosis)是真核細胞自然死亡的方式，個體藉細胞凋亡可除去受傷、老化或不要的細胞，以維持生物體的發育和細胞恆定。絞股藍是一種兼俱扶正補益與清熱解毒作用的抗癌中草藥，其萃取物絞股藍總皂甘有保肝、抗癌等廣泛功能，臨床上常用於治療肝癌的中醫複方中。本實驗目的即以體外實驗探討絞股藍總皂甘是否能誘發肝癌細胞株產生細胞凋亡的作用，並觀察是否能降低初代培養的人體周邊淋巴細胞凋亡和死亡率。 培養癌化程度不同之肝癌細胞株HepG2及HA22T/VGH，給予不同濃度的絞股藍總皂甘，培養不同時間，經MTT生長抑制試驗、電泳觀察DNA裂解片斷、流式細胞儀觀察PI染色後的細胞周期現象，分析是否有誘發細胞凋亡現象。抽血培養一名B型肝炎帶原患者的周邊淋巴細胞CD4，運用Annexin V方法比較加絞股藍總皂甘藥物24小時後體外培養淋巴細胞的凋亡及死亡率。 結果顯示絞股藍總皂甘對HepG2及HA22T/VGH均有生長抑制以及細胞周期有G1 arrest現象，電泳則見HA22T/VGH有DNA裂解片斷。體外培養淋巴細胞則顯示加絞股藍皂甘組對CD4細胞的凋亡及死亡率，較不加藥來得低，但加B型肝炎表面抗原組即使絞股藍皂甘同時存在下亦無降低。實驗支持絞股藍有誘導肝癌細胞株HepG2及HA22T/VGH細胞凋亡作用，對體外培養的周邊淋巴CD4細胞無毒害作用。; Eukaryotic cells normally expire through a natural process called apoptosis. This process is crucial for individuals to maintain their development and homeostasis by eliminating damaged, aged or unwanted cells. Gynostemma pentaphyllum is a Chinese medical herb possessing anti-tumor effects as it helps to restore the body’s overall balance, to enhance physiological performance, and has detoxification and antioxidant qualities. The active compounds present in the extract of Gynostemma pentaphyllum was found to be gypenosides which has wide range of actions and may help to promote liver functions and inhibit tumor growth. Gypenosides are therefore often included in medications for treating liver cancers. In this study, gypenosides were tested to induce apoptosis in cells of several hepatocellular carcinoma cell lines, and on other hand to protect the primary culture of human peripheral lymphocytes from apoptosis and cell death. Two hepatocellular carcinoma cell lines, HepG2 which is well differentiated, and HA22T/VGH, which was poorly differentiated, were cultured. After the addition of different concentrations of gypenosides, the cultures were incubated further for various periods of time. Induction of apoptosis was manifested by cytotoxicity in MTT tests, DNA fragmentation after electrophoresis and cell cycle arrest in flow cytometry analysis. For the evaluation of apoptosis and death rate of peripheral CD4 lymphocytes, primary culture was derived from blood specimen of a hepatitis B carrier. Subsequent to the administration of gypenosides, Annexin V analysis was performed 24 hours later.
