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目的探讨色素上皮衍生因子(PEDF)调控激素敏感性脂肪酶(HSL)抑制肥胖状态下脂肪动员的作用及机制。方法构建高脂喂养的肥胖小鼠模型,应用Western blot和荧光定量PCR检测PEDF和HSL的蛋白及mRNA水平;腹腔注射异丙肾上腺素(ISO)模拟激素诱导脂肪动员过程,应用Western blot检测HSL的磷酸化水平;培养3T3-L1并诱导分化为脂肪细胞,分别加ISO和AMPK激动剂利用Western blot研究PEDF调控HSL的分子机制。结果Western blot和荧光定量PCR检测结果提示高脂组PEDF蛋白水平和mRNA含量在各个时间点(4、8、12、16、20周)较普通组均明显上调,而HSL蛋白和mRNA水平在各个时间点较普通组均显著下调。在ISO诱导下,不同浓度(10、100、500 nmol/L)PEDF显著下调HSL磷酸化水平。Western blot检测提示PEDF可以同时下调脂肪细胞AMPK和HSL磷酸化水平。结论 PEDF可能通过抑制AMPK的磷酸化调控HSL,导致肥胖状态下体内甘油三酯进一步蓄积,最终诱导胰岛素抵抗等代谢异常疾病。
Objective To investigate the effect of pigment epithelium-derived factor (PEDF) on the regulation of lipid-sensitive lipase (HSL) on fat mobilization in obese rats. Methods The obese mouse model of high fat diet was constructed. The protein and mRNA levels of PEDF and HSL were detected by Western blot and real-time PCR. Intraperitoneal injection of isoprenaline (ISO) simulated hormones induced fat mobilization. Western blot was used to detect the expression of HSL Phosphorylation; cultured 3T3-L1 and induced differentiation into adipocytes, ISO and AMPK agonists were added respectively by Western blot PEDF molecular mechanism of HSL regulation. Results The results of Western blot and real-time PCR indicated that PEDF protein and mRNA levels in high-fat diet group were significantly increased at all time points (4, 8, 12, 16 and 20 weeks) The time points were significantly lower than the normal group. Under the induction of ISO, PEDF at different concentrations (10, 100 and 500 nmol / L) significantly down-regulated HSL phosphorylation. Western blot showed that PEDF can down-regulate the phosphorylation of AMPK and HSL in adipocytes at the same time. Conclusions PEDF may regulate HSL by inhibiting the phosphorylation of AMPK, resulting in further accumulation of triglycerides in the body during obesity and eventually inducing metabolic disorders such as insulin resistance.