五甲基槲皮素“棕色化”白色脂肪细胞及成年小鼠心脏c-kit细胞研究-study on.docx

华 华 中 科 技 大 学 博 士 论 文 PAGE VI PAGE VI Abstract Pentamethylquercetin browns the white adipocytes and the study of adult mouse cardiac c-kit+ cells PH.D Candidate: Han Yi Supervisor: Prof. Jin Manwen Department of Pharmacology, Tongji Medical College of Huazhong University of Science and Technology Part I. Pentamethylqercetin browns white adipocytes I. PMQ browns 3T3-L1 adipocytes Object: To study the effects of PMQ on 3T3-L1 adipocytes. Method: MTT, Oil red O stain, Janus Green B stain, the commercial kits of Glu, TG, and Glycerol, RT-PCR, Western blot and Realtime PCR were used to evaluate the effects of PMQ on the proliferation, differentiation, glucose consumption, cellular lipid metabolism, Mitochondria biogenesis, and the expression of related brown genes in 3T3-L1 adipocytes. Results: PMQ had no effect on the proliferation of 3T3-L1 preadipocytes. However, PMQ treatment upregulated the consumption of glucose, inhabited the accumulation of cellular TG, induced the biogenesis of Mitochondria and expression of UCP-1, PGC-1a, C/EBP a, PRDM16, PPAR?, PPARa. Conclus ion: PMQ can brown 3T3-L1 adipocytes, and affect on the early stage of adipocyte differentiation. II. Effects of PMQ on adult Kunming mice Object: To study the effects of PMQ on adult Kunming mice. Methods: 45 adult Kunming mice were randomized into Vehicle, PMQ 2.5, 5, 10 mg/kg and RSG 5 mg/kg group. Body weight s and food intakes were monitored weekly. After 14 days of PMQ treatment, the animals were sacrificed. Brown adipose tissues were weighed, and the expression of brown genes in WAT was analysed. Results: PMQ treatment for a short time had no impact on body weights, food intakes and BAT weights of adult Kunming mice, however the mRNA levels of UCP-1, PGC-1a, and C/EBPa were upregulated in WAT. III. Effects of PMQ on high-fat diet fed C57BL/6 mice Object: To explore the effects of PMQ on high- fat diet fed C57BL/6 mice. Method: 36 C57BL/6 mice were randomized