obestatin对3T3-L1前脂肪细胞增殖分化的影响及机制探讨
发布时间：2019-02-13 09:55
【摘要】： 目的观察obestatin对3T3-L1前脂肪细胞增殖分化的影响并探讨其作用机制。方法应用不同浓度的obestatin(10-8mmol/l、10-9mmol/l、10-10mmol/l、10-11mmol/l、10-12 mmol/l)干预体外培养的3T3-L1前脂肪细胞,用MTT法和葡萄糖消耗实验观察其对于细胞增殖的影响;应用10-10mmol/l obestatin全程干预3T3-L1前体脂肪细胞分化成熟过程(第0～10天),采用油红O染色法鉴定脂肪细胞分化并测定分化成熟脂肪细胞的脂肪含量;采用RT-PCR技术检测成熟脂肪细胞中PPARγ2基因mRNA的表达水平;采用RT FQ-PCR技术检测脂肪细胞中PTP1B基因mRNA的表达水平;采用Western-blot技术检测成熟脂肪细胞中PTP1B蛋白的表达水平;采用免疫沉淀和Western-blot技术检测脂肪细胞胰岛素受体磷酸化程度。结果①与对照组相比,不同浓度obestatin组细胞在570nm的吸光度值均有降低;②被10-10mmol/l obestatin连续作用10天的3T3-L1前脂肪细胞较对照组相比,脂肪产量明显减少;③脂肪细胞分化过程中PPARγ2基因表达逐渐增多,与对照组相比,obestatin组PPARγ2基因表达显著降低;④脂肪细胞分化过程中PTP1B基因表达逐渐减少,与对照组相比,obestatin组PTP1B基因和蛋白表达显著升高;⑤与对照组相比,obestatin组胰岛素受体磷酸化程度表达降低。 结论obestatin抑制3T3-L1前脂肪细胞的增殖和分化,其抑制作用可能是通过上调PTP1B表达继而抑制胰岛素信号传导通路实现的。
[Abstract]:Objective to observe the effect of obestatin on the proliferation and differentiation of 3T3-L1 preadipocytes and to explore its mechanism. Methods different concentrations of obestatin (10-8 mmol / L 10 -9 mmol / L 10 -10 mmol / L 10 -11 mmol / L 10 -12 mmol/l) were used to interfere with preadipocytes cultured in vitro. The effects of MTT and glucose consumption on cell proliferation were observed. 10-10mmol/l obestatin was used to interfere with the differentiation and maturation of 3T3-L1 precursor adipocytes (10 days). Oil red O staining was used to identify the differentiation of adipocytes and determine the fat content of mature adipocytes. The expression of PPAR 纬 2 gene mRNA in adipocytes was detected by RT-PCR, the expression of PTP1B gene mRNA in adipocytes was detected by RT FQ-PCR technique, the PTP1B protein expression level in adipocytes was detected by Western-blot technique. Insulin receptor phosphorylation in adipocytes was detected by immunoprecipitation and Western-blot. Results 1Compared with the control group, the absorbance of 570nm was decreased in different concentrations of obestatin group, and the fat production of 3T3-L1 cells exposed to 10-10mmol/l obestatin for 10 days was significantly lower than that of control group. 3The expression of PPAR 纬 2 gene increased gradually during adipocyte differentiation. Compared with the control group, the expression of PPAR 纬 2 gene in obestatin group was significantly lower than that in control group. (4) the expression of PTP1B gene decreased gradually during adipocyte differentiation. Compared with the control group, the expression of PTP1B gene and protein in obestatin group was significantly increased, and the expression of insulin receptor phosphorylation in obestatin group was lower than that in control group. Conclusion obestatin inhibits the proliferation and differentiation of 3T3-L1 preadipocytes by up-regulating the expression of PTP1B and inhibiting insulin signaling pathway.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
本文编号：2421426
[Abstract]:Objective to observe the effect of obestatin on the proliferation and differentiation of 3T3-L1 preadipocytes and to explore its mechanism. Methods different concentrations of obestatin (10-8 mmol / L 10 -9 mmol / L 10 -10 mmol / L 10 -11 mmol / L 10 -12 mmol/l) were used to interfere with preadipocytes cultured in vitro. The effects of MTT and glucose consumption on cell proliferation were observed. 10-10mmol/l obestatin was used to interfere with the differentiation and maturation of 3T3-L1 precursor adipocytes (10 days). Oil red O staining was used to identify the differentiation of adipocytes and determine the fat content of mature adipocytes. The expression of PPAR 纬 2 gene mRNA in adipocytes was detected by RT-PCR, the expression of PTP1B gene mRNA in adipocytes was detected by RT FQ-PCR technique, the PTP1B protein expression level in adipocytes was detected by Western-blot technique. Insulin receptor phosphorylation in adipocytes was detected by immunoprecipitation and Western-blot. Results 1Compared with the control group, the absorbance of 570nm was decreased in different concentrations of obestatin group, and the fat production of 3T3-L1 cells exposed to 10-10mmol/l obestatin for 10 days was significantly lower than that of control group. 3The expression of PPAR 纬 2 gene increased gradually during adipocyte differentiation. Compared with the control group, the expression of PPAR 纬 2 gene in obestatin group was significantly lower than that in control group. (4) the expression of PTP1B gene decreased gradually during adipocyte differentiation. Compared with the control group, the expression of PTP1B gene and protein in obestatin group was significantly increased, and the expression of insulin receptor phosphorylation in obestatin group was lower than that in control group. Conclusion obestatin inhibits the proliferation and differentiation of 3T3-L1 preadipocytes by up-regulating the expression of PTP1B and inhibiting insulin signaling pathway.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
【参考文献】
相关期刊论文 前1条
1 杨凌辉,邹大进;肥胖致胰岛素抵抗的机制[J];中华内分泌代谢杂志;2002年03期
，本文编号：2421426
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2421426.html
最近更新
教材专著
