【摘要】： 目的觀察obestatin對(duì)3T3-L1前脂肪細(xì)胞增殖分化的影響并探討其作用機(jī)制。方法應(yīng)用不同濃度的obestatin(10-8mmol/l、10-9mmol/l、10-10mmol/l、10-11mmol/l、10-12 mmol/l)干預(yù)體外培養(yǎng)的3T3-L1前脂肪細(xì)胞,用MTT法和葡萄糖消耗實(shí)驗(yàn)觀察其對(duì)于細(xì)胞增殖的影響;應(yīng)用10-10mmol/l obestatin全程干預(yù)3T3-L1前體脂肪細(xì)胞分化成熟過(guò)程(第0～10天),采用油紅O染色法鑒定脂肪細(xì)胞分化并測(cè)定分化成熟脂肪細(xì)胞的脂肪含量;采用RT-PCR技術(shù)檢測(cè)成熟脂肪細(xì)胞中PPARγ2基因mRNA的表達(dá)水平;采用RT FQ-PCR技術(shù)檢測(cè)脂肪細(xì)胞中PTP1B基因mRNA的表達(dá)水平;采用Western-blot技術(shù)檢測(cè)成熟脂肪細(xì)胞中PTP1B蛋白的表達(dá)水平;采用免疫沉淀和Western-blot技術(shù)檢測(cè)脂肪細(xì)胞胰島素受體磷酸化程度。結(jié)果①與對(duì)照組相比,不同濃度obestatin組細(xì)胞在570nm的吸光度值均有降低;②被10-10mmol/l obestatin連續(xù)作用10天的3T3-L1前脂肪細(xì)胞較對(duì)照組相比,脂肪產(chǎn)量明顯減少;③脂肪細(xì)胞分化過(guò)程中PPARγ2基因表達(dá)逐漸增多,與對(duì)照組相比,obestatin組PPARγ2基因表達(dá)顯著降低;④脂肪細(xì)胞分化過(guò)程中PTP1B基因表達(dá)逐漸減少,與對(duì)照組相比,obestatin組PTP1B基因和蛋白表達(dá)顯著升高;⑤與對(duì)照組相比,obestatin組胰島素受體磷酸化程度表達(dá)降低。 結(jié)論obestatin抑制3T3-L1前脂肪細(xì)胞的增殖和分化,其抑制作用可能是通過(guò)上調(diào)PTP1B表達(dá)繼而抑制胰島素信號(hào)傳導(dǎo)通路實(shí)現(xiàn)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 緯 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 緯 2 gene increased gradually during adipocyte differentiation. Compared with the control group, the expression of PPAR 緯 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.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 楊凌輝,鄒大進(jìn);肥胖致胰島素抵抗的機(jī)制[J];中華內(nèi)分泌代謝雜志;2002年03期

，

本文編號(hào)：2421426