ATP敏感性钾通道对大鼠前脂肪细胞增殖分化影响及其机制

发布时间：2018-05-30 13:26

本文选题：大鼠 + 钾通道　；参考：《华中科技大学》2007年硕士论文

【摘要】： 一、ATP敏感性钾通道对大鼠前脂肪细胞增殖分化影响目的:探讨ATP敏感性钾通道在大鼠前脂肪细胞增殖分化中作用。方法:逆转录PCR检测大鼠脑组织、脂肪组织、前脂肪细胞和诱导5天获得的脂肪细胞中ATP敏感性钾通道磺脲类受体mRNA (SURmRNA)亚型,实时定量PCR检测前脂肪细胞和诱导5天获得的脂肪细胞中SURmRNA表达;ATP敏感性钾通道阻滞剂格列本脲和激动剂二氮嗪作用于前脂肪细胞,实时定量PCR检测SURmRNA表达,MTT检测前脂肪细胞增殖,流式细胞仪检测细胞周期分布;油红O染色法检测细胞内脂质含量,Image-Pro plus5.0软件测量细胞直径。结果:大鼠脑组织中有SUR1mRNA、SUR2mRNA表达,大鼠脂肪组织、前脂肪细胞和诱导5天获得的脂肪细胞中均有SUR2mRNA表达,而无SUR1mRNA表达,且脂肪细胞中SUR2mRNA表达明显高于前脂肪细胞;格列本脲抑制前脂肪细胞SUR2mRNA表达,呈剂量依赖性地促进前脂肪细胞增殖,增加G2/M+S期细胞百分率,增加细胞脂质含量,使脂肪细胞直径增大;二氮嗪在这些方面的作用与格列本脲相反。结论:ATP敏感性钾通道在前脂肪细胞增殖分化中可能起调节作用。二、ATP敏感性钾通道调节大鼠前脂肪细胞增殖分化机制研究目的:探索ATP敏感性钾通道调节大鼠前脂肪细胞增殖分化机制。方法:ATP敏感性钾通道特异性激动剂(二氮嗪)、阻滞剂(格列本脲)作用于前脂肪细胞诱导分化5天后,逆转录PCR检测瘦素mRNA表达及过氧化物酶体增殖物激活受体γ(PPARγ)mRNA表达。二氮嗪、格列本脲作用于前脂肪细胞一天,Western blot检测其细胞内P21、P27蛋白表达。结果:前脂肪细胞分化过程中,二氮嗪组瘦素mRNA表达明显增加(p0.05),格列本脲组瘦素mRNA表达明显降低(p0.05);格列本脲组PPARγmRNA表达增加(p0.05),而二氮嗪组PPARγmRNA表达减少(p0.05)。二氮嗪作用于前脂肪细胞,使细胞内P21、P27表达增高(p0.05),而格列本脲作用于前脂肪细胞后,细胞内P21、P27表达降低(p0.05)。结论:ATP敏感性钾通道调节大鼠前脂肪细胞增殖分化,可能与瘦素mRNA、P21、P27蛋白表达上调,PPARγmRNA表达下调有关
[Abstract]:Effects of ATP-sensitive potassium channels on proliferation and differentiation of rat preadipocytes Aim: to investigate the role of ATP sensitive potassium channel in the proliferation and differentiation of rat preadipocytes. Methods: reverse transcription PCR was used to detect the subtypes of ATP sensitive potassium channel sulfonylurea receptor (mRNA) in rat brain, adipocytes, preadipocytes and adipocytes induced for 5 days. Real-time quantitative PCR was used to detect SURmRNA expression in preadipocytes and adipocytes obtained after 5 days of induction. Glibenclamide, an ATP-sensitive potassium channel blocker, and diazazine, a agonist, acted on preadipocytes. The expression of SURmRNA was detected by real-time quantitative PCR, the proliferation of adipocytes was detected by flow cytometry, the lipid content in cells was detected by oil red O staining and the diameter of adipocytes was measured by Image-Pro plus5.0 software. Results: the expression of SUR1mRNA-SUR2 mRNA was found in rat brain tissue. The expression of SUR2mRNA was found in adipose tissue, preadipocytes and adipocytes induced for 5 days, but no SUR1mRNA expression was found in adipocytes. The expression of SUR2mRNA in adipocytes was significantly higher than that in preadipocytes. Glibenclamide inhibited the expression of SUR2mRNA in preadipocytes, promoted the proliferation of preadipocytes in a dose-dependent manner, increased the percentage of G2 / MS phase cells, increased the lipid content and increased the diameter of adipocytes. Diazazine has the opposite effect on these aspects as glibenclamide. Conclusion the cell proliferation and differentiation of preadipocytes may be regulated by K 2 + channel. Mechanism of ATP-sensitive potassium channels regulating proliferation and differentiation of rat preadipocytes Aim: to explore the mechanism of ATP sensitive potassium channel regulating the proliferation and differentiation of preadipocytes in rats. Methods expression of leptin mRNA and peroxisome proliferator activated receptor 纬 -PPAR 纬 -PPAR 纬 mRNA were detected by reverse transcriptase PCR (RT PCR) after differentiation of preadipocytes was induced by diazosin and glibenclamide (glibenclamide). Preadipocytes were treated with diazazine and glibenclamide for one day. Western blot was used to detect the expression of P21 P27 protein in preadipocytes. Results: during the differentiation of preadipocytes, the expression of leptin mRNA increased significantly in diazazine group, the leptin mRNA expression in glibenclamide group decreased significantly, and the expression of PPAR 纬 mRNA increased in glibenclamide group, while PPAR 纬 mRNA expression decreased in diazazine group. After treated with diazoxide, the expression of P21 P27 in preadipocytes increased, while glibenclamide decreased the expression of P21 P27 in preadipocytes. Conclusion the proliferation and differentiation of preadipocytes regulated by the potassium channel of 1: ATP-sensitive may be related to the up-regulation of the expression of leptin mRNA-P21p27 protein and down-regulation of the expression of PPAR- 纬 mRNA in rat preadipocytes.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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