| Summary: | 碩士 === 弘光科技大學 === 營養醫學研究所 === 101 === Aristolochic acid (AA), derived from the plants of Aristolochia species, has been documented to be associated with aristolochic acid nephropathy (AAN) and tumors. AA used to be an ingredient of some health supplements and herbal medicine all over the world. It has been documented in epidemiology, cellular, animal studies that AA is associated with increased apoptosis of renal tubular cell and raise of reactive oxygen species (ROS). The deterioration of renal function associated with aristolochic acid could be fast. After taking, renal function may be decreased in a short period and even progress to end-stage renal disease(ESRD) which dialysis is instituted. As mention in some studies, Aristolochic acid induces damage of proximal renal tubular cell, fibrosis, epithelial- mesenchymal transition(EMT), and even carcinogenesis. Apoptosis of renal tubular cells could be attenutated by antioxidant in some studies.
In our study, renal tubular cells (NRK-52E) are incubated either with or without AA at various concentration(0 ,10 ,20 and 100 µM) and various times (6 , 12, 24 and 48 hours) for documenting the cell death and raise of ROS. The results are the viable rate of NRK-52E declined and production of ROS increased as the rise of AA concentration or longer time of incubation at every specific concentration of AA (p<0.05). When NRK-52E cells incubated with 10 µM AA, various concentration of vitamin C (0 , 2.5 , 5 , 10 and 50µM) were added. The results disclose vitamin C at low concentration (5 μM) may decrease the cell death and production of ROS by 10 µM AA . Compaired to non-vitamin C, high concentration of vitamin C (50μM) decreased viable rate of NRK-52E instead of improving when co-culturing with 10 µM AA (p<0.05). Analysis of NRK-52E cell cultured in 0 , 10 and 20 µM AA by H33342 showed the character of cell death is condensation of nucleus inducing apoptosis, In cells with 0 µM AA and 10µM AA with or without 5 µM vitamin C, cleaved-caspase 3 protein increased in group of 10 µM AA than 0 µM and decrease in group of 10 µM AA with 5 µM vitamin C than without. In further analysis by H33342 showed number of nucleus condensation decreased at 5 µM vitamin C compared to 0 µM vitamin C when NRK-52E cells co-cultured with 10µM AA.
In conclusion, when NRK-52E co-culture with vitamin C and AA, cell apoptosis and production of ROS induced by AA may be reduced.
Keywords: Vitamin C, Aristolochic acid, Apoptosis, NRK-52E
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