| Summary: | 碩士 === 中原大學 === 生物醫學工程研究所 === 101 === Along with unbalance diet and sedentary lifestyle, the population of diabetes mellitus is increasing significantly. Puerarin, a major active ingredient extracted from Pueraria lobata, is known as antihyperglycemic and antioxidantive agents. However, the poor oral availability and aqueous solubility are major barriers for drug formulation. Therefore, to effectively deliver puerarin, a liposomal dosage form of puerarin is used. This study aimed to develop a stable and biocompatible liposomal formulation for combating diabetic nephropathy disease. Two-phase solvent were used to extract the puerarin at room temperature、50 oC and 70 oC, the purity was found to be 83.4、70.5 and 55.6 % respectively. The anti-oxidative capacity of puerarin were evaluate by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical experiment, and found 1.25 mg/mL extracted puerarin can eliminate up to 22.1 % of free radical (DPPH) in the first ten minutes of incubation. Liposomes (E70C30T0、E65C30T5、E60C30T10) was prepared by water-bath sonicaion or membrane extrution methods and characterized for the particle size and its stability in sucrose and rabbit serum solution. In sucrose solution for 5 days, increase ratio of the particle was 11.97、1.07 and 1.94 % by sonication method, and 1.88、1.78 and 1.55 % by extrution method, respectively. It was found that membrane extrution of E60C30T10 formulation could produce the stable particles in sucrose solution. However, the phosphate groups of liposomes formula in serum was significantly hydrolyzed by serum carboxyesterase. In the third part, liposomal puerarin were prepared using extrution method (E70C30T0、E65C30T5、E60C30T10). The drug loading efficiency were 40.45、49.24 and 53.79 %, respectively. The study of in vitro cytotoxicity showd the puerarin encapsulated by liposomes could increased 1.19 times of cell viability of mesangial cell line compared with free puerarin. The 33 mM glucose medium significantly encanced the reactive oxygen species (ROS) level, and improve the cell proliferation, which was used as an in vitro model of diabetic nephropathy. The cell proliferation、gene expression of TGF-β1 and nuclear translocation of Smad 2/3 proteins in 33 mM glucose medium were suppressed in a concentration dependent manner by liposomal puerarin. In summary, this study successfully prepare the liposomal puerarin wich can provide cytoprotective effect in high glucose condition, and is of potential in combating diabetic nephropathy.
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