| Summary: | 碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 102 === Ilex kudingcha, also known as a leaf tea, is a traditional beverage found to have hypoglycemic and hepato-protective functions. Alternanthera philoxeroides (Mart.) Griseb, also called alligator weed, originated in South America that exhibits heat-purging and antibacterial effects. However, immunomodulatory effects of Ilex kudingcha and Alternanthera philoxeroides remain unclear. To clarify the puzzle, Ilex kudingcha and Alternanthera philoxeroides were extracted using three different solvents, including chloroform, methanol and water, respectively. The extracts were first subjected to Th1/Th2 immunomodulatory study using mouse primary splenocytes in vitro. Two potential extracts were isolated to explore their anti-inflammatory and anti-allergic effects using mouse primary peritoneal macrophages and lung mast cells, respectively. Finally, potent extracts were further selected to treat human prostate adenocarcinoma PC-3 cells and breast adenocarcinoma MCF-7 cells using direct addition or natural killer cells’ immunotherapy.
The results showed that Ilex kudingcha chloroform extract (IKCE) had potential to regulate Th1/Th2 immune responses toward Th2 immune balance and Ilex kudingcha methanol extract (IKME) toward Th1 immune balance among six different extracts.
Therefore, IKCE and IKME were screened to explore their anti-inflammatory effects using mouse peritoneal macrophages. IKCE and IKME treatments alone showed a mild stimulatory activity to macrophages, suggesting that both extracts might have potential to enhance innate immunity. IKCE and IKME treatments in the presence of lipopolysaccharides(LPS) were found to have anti-inflammatory potential. Furthermore, both IKCE and IKME had prophylactic effects against LPS-induced inflammation through inhibiting pro-/anti-inflammatory cytokine secretion ratios by peritoneal macrophages in a specified preventive experiment. In addition, IKCE and IKME inhibited the degranulation of mouse lung mast cells, suggesting that both extracts might have anti-allergic potential.
IKCE and IKME were suggested to have potential to enhance innate immunity; therefore IKCE and IKME were further selected to treat PC-3 and MCF-7 cells using direct addition or the conditioned medium of natural killer cells (NK92-MI) cultured with there potent extracts.The results showed that IKME inhibited the proliferation of human prostate adenocarcinoma PC-3 cells. Consequently, IKME and its conditioned medium of NK92-MI cells were further selected to treat PC-3 cells. Changes of mRNA expression levels of pro- (Bax)/anti-apoptotic(Bcl-2) genes in PC-3 cells were assayed using RT-qPCR. Results showed that expression ratios of Bax/Bcl-2 mRNA levels in PC-3 cells were significantly increased by IKME and its conditioned medium of NK92-MI cells. In addition, Fas gene expression levels in PC-3 cells were also increased by IKME and its conditioned medium of NK92-MI cells.
In conclusion, results from this study evidenced that IKME treatments might induce apoptosis in PC-3 cells through increasing pro-apoptotic Bax and Fas genes expression via
direct action or natural killer cells’ immunotherapy to achieve its anticancer effects.
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