부산대학교 도서관

ABSTRACTAcorus gramineus Soland. is a shrubby plant which belongs to Araceae family and mostly distributes in Korea, China, Japan, Indonesia, Philippines and India. Acorus gramineus Soland. rhizome has been widely used as a traditional medicine in China and Korea. However, there are just a few researchers studying its effects on anti-obesity. This paper aims at evaluating its inhibitory effects on 3T3-L1 preadipocyte differentiation with 70% ethanol and hot water extract In-vitro and anti-obesity effects in C57BL/6N mice both with powder and 70% ethanol extract In-vivo. Acorus gramineus Soland. rhizome was purchased in Korea, and extracted with 70% ethanol (AGE) and hot water (AGW) for further experiments. In present experimental work, the proximate composition (crude protein, crude fat, crude ash and carbohydrate contents) and total dietary fiber content of Acorus gramineus Soland. rhizome were investigated. The nutrient component (total protein, free amino acid, total sugar and reducing sugar contents), total polyphenol content and antioxidant activity of Acorus gramineus Soland. rhizome extracts have also been studied. Results show that the largest proportion is the carbohydrate content (83.79%) in the proximate composition of Acorus gramineus Soland. rhizome. And in carbohydrate content, insoluble dietary fiber (53.77%) is the main proportion. Even though AGE extraction yield (18.78 %) was lower than that in AGW (30.48%), but AGE polyphenol content (35.11 mg/g) was showed higher than that in AGW (11.45 mg/g). AGE also indicated a potenter ABTs and DPPH radical scanvenging activities than AGW. The RC50 values of ABTs and DPPH radical scanvenging activities of AGE were 0.93 and 2.35 mg/mL, respectively.The anti-obesity activity In-vitro was evaluated by examining the pancreatic lipase, α-amylase inhibitory activity and inhibitory effects on 3T3-L1 preadipocyte differentiation. In addition, in order to confirm the anti-obesity molecular mechanisms of AGE, the protein and mRNA expression of adipogenic genes, such as peroxisome proliferator-activated receptor γ (PPARγ), CCATT/enhancer-binding protein α (C/EBPα), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS) and acetyl-Coenzyme A carboxylase (ACC) were determined by western blot and RT-PCR analysis. Further more, five fractions (n-Hexane, CH2Cl2, EtOAc, n-BuOH and H2O-soluble fractions) isolated from Acorus gramineus Soland. rhizome methanol extract were operated for the pancreatic lipase and α-amylase inhibitory activities, and studied the inhibitory effects on 3T3-L1 preadipocyte differentiation. In the anti-digestive enzyme activity test, AGE respectively inhibited lipase activity by 33.37% and α-amylase activity by 43.90%, which was investigated at the concentration of 10 mg/mL. In addition, n-Hexane, EtOAc, CH2Cl2 and n-BuOH-soluble fractions from MeOH extract respectively inhibited lipase activity by 82.22%, 78.02%, 62.63% and 28.61% at the same concentration of 8 mg/mL. The α-amylase activity was respectively inhibited by 71.18%, 66.79%, 64.09%, 63.15% and 57.20%, based on the order following by CH2Cl2, EtOAc, n-Hexane, n-BuOH and H2O-soluble fractions at the same concentration of 8 mg/mL. In the lipid accumulation inhibitory analysis, AGE inhibited the lipid accumulation in 3T3-L1 preadipocytes with 40.09% at the concentration of 0.60 mg/mL. AGE significantly suppressed preadipocyte differentiation by down regulating the genes of PPARγ, C/EBPα, SREBP-1c, FAS and ACC in a dose-dependent manner in transcription and translation levels. CH2Cl2-soluble fraction, as an active fraction isolated from Acorus gramineus Soland. rhizome, showed a significant inhibitory effects on lipid accumulation by 71.54% at the concentration of 0.10 mg/mL compared with MDI-treated cells. In-vivo, the anti-obesity effects of Acorus gramineus Soland. rhizome in C57BL/6N animal model were investigated. AG powder (0.25%, 0.5%, 1%) and AG ethanol extract (0.05%, 0.1%, 0.2%) were filled into high-fat diet during 8 weeks in present experiment. Supplementation of AGP and AGE with a dose-dependent manner significantly decreased the body weight gain, adipose tissues weight gain, increase of lipid levels (TG, TC levels in serum and liver), increase of glucose levels and also reduced the disordered hormones such as insulin and leptin levels caused by high-fat diet. In addition, it significantly increased the decrease of adiponectin levels induced by high-fat diet. We also found that AGP supplementation was more efficient than AGE supplementation in the anti-obesity effects In-vivo. Finally, the protein and mRNA expression of PPARγ, C/EBPα, SREBP-1c, FAS and ACC in liver were investigated by western blot and RT-PCR analysis. Similar to results In-vitro, AGP and AGE supplementation significantly inhibited the expression of PPARγ, C/EBPα, SREBP-1c, FAS and ACC in hepatic tissue in transcription and translation levels.Above results demonstrate that Acorus gramineus Soland. rhizome has strongly potent effects on anti-obesity In-vitro and In-vivo. Acorus gramineus Soland. rhizome contributes to the development of new nature resources drugs to treat obesity. Moreover, Acorus gramineus Soland. rhizome can be regarded as a potential drug for treat type-2 diabetes mellitus.