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Exendin-4对脂肪细胞分化及相关基因表达的影响
Effects of Exendin-4 on Adipocyte Differentiation Related Gene Expression of Adipocyte Markers Gene
【作者】 赵向；
【导师】 肖新华；
【作者基本信息】 南华大学 ， 内分泌， 2010， 硕士
【摘要】 目的:探讨Exendin-4对3T3-L1前脂肪细胞分化及GLUT-4、PPAR-γ、HSL mRNA表达的影响。方法:3T3-L1前脂肪细胞诱导分化不同时期分别用Exendin-4等干预,应用倒置相差显微镜观察3T3-L1前脂肪细胞诱导分化前后的细胞形态变化;油红O染色后观察脂滴数量;Q-PCR分别检测GLUT-4、PPAR-γ、HSL mRNA表达量;应用酶法测定脂肪细胞的甘油三酯(TG)含量。结果:诱导分化成熟的脂肪细胞经油红O染色可见细胞质内大片脂滴呈亮红色,而未分化细胞则不被油红O染色。在脂肪细胞分化第0d干预,Exendin-4可促进脂肪细胞分化,较空白组增加细胞内TG的含量(P<0.01),上调GLUT-4、PPAR-γ、HSL mRNA的表达(P<0.01),TNF-α组较空白组均能显著下调GLUT-4、HSL mRNA的表达(P<0.01);与Troglitazone组比较,EX-4组PPAR-γmRNA的表达降低(P<0.05),EX-4+Trogli组GLUT-4、HSL mRNA的表达显著增加(P<0.01),EX-4+Trogli组PPAR-γmRNA的表达增加(P<0.05)。在脂肪细胞分化6d干预,Exendin-4可促进脂肪细胞分化,较空白组增加细胞内TG的含量(P<0.01),上调GLUT-4、HSL、PPAR-γmRNA的表达(P<0.01);TNF-α组较空白组能显著下调GLUT-4、HSL mRNA的表达(P<0.01),下调PPAR-γmRNA的表达(P<0.05);与Troglitazone组比较,EX-4组GLUT-4 mRNA的表达降低(P<0.05),HSL mRNA的表达显著降低(P<0.01),EX-4+Trogli组HSL、PPAR-γmRNA的表达增加(P<0.05)。脂肪细胞分化第12d干预,Trogli组、EX-4组+Trogli组较空白组能上调GLUT-4 mRNA的表达(P<0.05);TNF-α组较空白组能下调HSL mRNA的表达(P<0.01);与Troglitazone组比较,EX-4组GLUT-4、HSL mRNA的表达减少(P<0.05),EX-4组+Trogli组HSL mRNA的表达增加(P<0.05),TG含量也增加(P<0.05)。结论:Exendin-4促进脂肪细胞分化并上调糖脂代谢相关基因GLUT-4、PPAR-γ、HSL mRNA表达,可能为Exendin-4抗糖尿病的部分作用新机制。
【Abstract】 Objective: To investigate the effects of Exendin-4 on differentiation of 3T3-L1 preadipocytes and mRNA expressions of GLUT-4、PPAR-γand HSL.Methods: 3T3-L1 preadipocytes of different differentiation stages were treated by Exendin-4. Morphological changes of 3T3-L1 preadipocytes and cell differentiation were measured by inverted phase contrast mocroscope. The number of lipid droplets was observed by Oil red O staining. The mRNA expressions of GLUT-4、PPAR-γand HSL were detected by quantitative real-time polymerase chain reaction(RT-PCR).The triglyceride content of 3T3-L1 preadipocytes was quantitated with an acetyl acetone–based colorimetric kit.Results: Induced differentiation of mature adipocytes showed large lipid droplets of red within the cytoplasm by oil red O staining, undifferentiated cells are not stained by oil red O.Comparing with control group, Exendin-4 promoted preadipocyte differentiation, increased TG content (P<0.01) and the mRNA expression of GLUT-4, HSL and PPAR-γ(P<0.01) when treated with Exendin-4 at day 0.Compared with the control group,TNF-αsignificantly reduced mRNA expressions of GLUT-4 and HSL (P<0.01). Compared with troglitazone group, the expressions of PPAR-γmRNA reduced in EX-4 group (P<0.05), the expressions of GLUT-4 and HSL mRNA significantly increased (P<0.01) and the expressions of PPAR-γmRNA increased (P<0.05) in Ex-4 combined with troglitazone group.Comparing with control group, Exendin-4 promoted preadipocyte differentiation, increased TG content (P<0.01) and the mRNA expressions of GLUT-4, HSL andPPAR-γ(P<0.01) when treated with Exendin-4 at day 6. Compared the control group, TNF-αgroup significantly reduced GLUT-4 and HSL mRNA expressions (P<0.01) and reduced PPAR-γmRNA expression (P<0.05); Compared with Troglitazone group , the expressions of GLUT-4 mRNA reduced in Ex-4 group (P<0.05) and the expression of HSL mRNA significantly reduced (P<0.01). Compared with Troglitazone group, the expressions of HSL and PPAR-γmRNA increased in Ex-4 combined with troglitazone group (P <0.05).Compared with control group, the expressions of GLUT-4 mRNA increased in Ex-4 combined with troglitazone group and Trogli group(P<0.05). Compared with control group, the expressions of HSL mRNA reduced in TNF-αgroup (P<0.01) when treated with Exendin-4 at day. Compared with Troglitazone group, the expressions of GLUT-4 and HSL mRNA reduced in Ex-4 group (P<0.05), Compared with Troglitazone group, increased TG content (P<0.01) and the mRNA expression of HSL mRNA increased in Ex-4 combined with troglitazone group (P<0.05) and TG content also increased (P<0.05).Conlusion: Exendin-4 promoted adipocyte differentiation,increased glucose and lipid metabolism related genes mRNA expression, which may be some new mechanisms of Exendin-4 anti-diabetic.