本文關(guān)鍵詞：肌肉肌醇、D-手性肌醇及二者合用對(duì)HepG2胰島素抵抗細(xì)胞葡萄糖消耗量的改善作用　出處：《青島大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

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【摘要】：目的通過(guò)建立Hep G2胰島素抵抗模型(Hep G2-IR),比較肌肉肌醇(myo-inositol,MI)、D-手性肌醇(D-chirol-inositol,DCI)及二者合用對(duì)Hep G2-IR細(xì)胞葡萄糖消耗量的影響,并探討其作用的可能機(jī)制。方法1.CCK-8法檢測(cè)不同濃度(0、25、35、45、55、65mmol/L)的葡萄糖對(duì)Hep G2細(xì)胞增殖活性的影響,不同濃度(0、0.25、0.5、1、2、4、8、16、32mmol/L)的MI、DCI及二者合用分別對(duì)Hep G2細(xì)胞增殖活性的影響,以確定其在實(shí)驗(yàn)中的作用劑量范圍。2.濃度為55mmol/L的葡萄糖持續(xù)作用Hep G2細(xì)胞24h后,1nmol/L胰島素刺激誘導(dǎo)Hep G2細(xì)胞15~25min,建立胰島素抵抗細(xì)胞模型;通過(guò)葡萄糖氧化酶法(GOD-POD法)檢測(cè)對(duì)照組與模型組葡萄糖消耗量,以鑒定模型是否成立。3.不同濃度(0.125、0.25、0.5mmol/L)的MI、DCI及二者合用分別作用Hep G2-IR細(xì)胞,GOD-POD法檢測(cè)它們對(duì)葡萄糖消耗量的影響。Western-blot法檢測(cè)MI、DCI及二者合用對(duì)Hep G2-IR細(xì)胞PI3K、p Akt、Akt蛋白表達(dá)的影響。結(jié)果1.與空白對(duì)照組相比,葡萄糖在35~55mmol/L濃度時(shí),對(duì)Hep G2細(xì)胞增殖活性無(wú)影響,而葡萄糖在濃度為65mmol/L時(shí),對(duì)Hep G2細(xì)胞增殖活性有顯著的抑制作用(P0.05);0~0.5mmol/L濃度的MI、DCI對(duì)Hep G2細(xì)胞增殖活性無(wú)影響,而1~32mmol/L濃度的MI、DCI對(duì)Hep G2細(xì)胞增殖活性有顯著的抑制作用(P0.01),且隨MI、DCI濃度的增加,細(xì)胞增殖活性有逐漸降低的趨勢(shì);0~1mmol/L濃度的MI與DCI合用對(duì)Hep G2細(xì)胞增殖活性無(wú)影響,而2~32mmol/L濃度的MI與DCI合用對(duì)Hep G2細(xì)胞增殖活性有顯著的抑制作用(P0.01),且隨MI與DCI合用濃度的增加,細(xì)胞增殖活性有逐漸降低的趨勢(shì)。2.GOD-POD法檢測(cè)結(jié)果表明,與正常對(duì)照組相比,濃度為55mmol/L的葡萄糖持續(xù)作用Hep G2細(xì)胞24h,且1nmol/L胰島素刺激15-25min后,被誘導(dǎo)的Hep G2細(xì)胞葡萄糖消耗量顯著降低(P0.01),表明成功建立了Hep G2-IR細(xì)胞模型。3.GOD-POD法檢測(cè)顯示,與模型對(duì)照組相比,MI、DCI及二者合用組都可增加Hep G2-IR細(xì)胞葡萄糖消耗量(P0.05),且每個(gè)組隨著干預(yù)濃度增加,葡萄糖消耗量有逐漸上升的趨勢(shì),在相同濃度下,合用組葡萄糖消耗量增加最顯著(P0.05)。4.Western-blot分析顯示,與模型對(duì)照組相比,MI、DCI及二者合用組可增加Hep G2-IR細(xì)胞PI3K蛋白表達(dá)及p Akt/Akt蛋白表達(dá)水平的比值,且每個(gè)組隨著干預(yù)濃度增加,蛋白表達(dá)及蛋白表達(dá)水平的比值有逐漸上升的趨勢(shì),在相同濃度下,合用組提高的PI3K蛋白表達(dá)及p Akt/Akt蛋白表達(dá)水平的比值最顯著(P0.05)。結(jié)論MI、DCI及二者合用可能通過(guò)提高Hep G2-IR細(xì)胞的PI3K蛋白表達(dá)及促進(jìn)Akt的磷酸化,進(jìn)而增加Hep G2-IR細(xì)胞葡萄糖消耗量,從而達(dá)到改善IR目的,其中合用作用效果最顯著。
[Abstract]:Objective to compare the muscle myo-inositol (MIM) of Hep G2 insulin resistance model (Hep G2-IRI). Effects of D-chirol-inositol (DCI) and its combination on glucose consumption in Hep G2-IR cells. The possible mechanism of its action was also discussed. 1. CCK-8 method was used to detect the different concentrations of 25, 25, 35, 45 and 55. The effect of 65 mmol / L glucose on the proliferation activity of Hep G2 cells was observed. The effects of DCI and their combination on the proliferation of Hep G2 cells were studied. In order to determine the range of dose of glucose in the experiment. The concentration of glucose was 55 mmol / L after 24 hours of continuous treatment of Hep G2 cells. 1 nmol / L insulin stimulated Hep G2 cells were induced for 15 to 25 min to establish insulin resistance cell model. The glucose consumption of the control group and the model group was determined by glucose oxidase method (GOD-POD) to determine whether the model was established or not. Hep G2-IR cells were treated with 0.5 mmol / L MIDCI and their combination. GOD-POD assay was used to detect their effect on glucose consumption. Western-blot was used to detect PI3K in Hep G2-IR cells. Results 1.Compared with the blank control group, glucose at the concentration of 355mmol / L had no effect on the proliferation activity of Hep G2 cells. When the concentration of glucose was 65 mmol / L, the proliferative activity of Hep G2 cells was inhibited significantly (P 0.05). 0.5 mmol / L MI-DCI had no effect on the proliferation activity of Hep G2 cells, while the MI of 32 mmol / L concentration was not affected. The proliferation activity of Hep G2 cells was significantly inhibited by DCI, and the proliferation activity decreased gradually with the increase of MI-DCI concentration. The combination of 0 mmol / L MI and DCI had no effect on the proliferation activity of Hep G2 cells. The proliferation activity of Hep G2 cells was significantly inhibited by the combination of MI and DCI at the concentration of 32 mmol / L, and increased with the increase of the concentration of MI and DCI. The proliferation activity of the cells decreased gradually. 2. The results of GOD-POD assay showed that compared with the normal control group. Glucose at a concentration of 55 mmol / L continuously stimulated Hep G2 cells for 24 h, and 1 nmol / L insulin was stimulated for 15-25 min. The glucose consumption of induced Hep G2 cells decreased significantly, indicating that the Hep G2-IR cell model was successfully established. 3. Compared with the model control group, the glucose consumption of Hep G2-IR cells was increased in both groups, and each group increased with the intervention concentration. Glucose consumption increased gradually in the same concentration, the glucose consumption of the combined group increased most significantly (P0.05N. 4. Western-blot analysis showed. Compared with the model control group, the PI3K protein expression and the ratio of p Akt/Akt protein expression in Hep G2-IR cells were increased. With the increase of intervention concentration, the ratio of protein expression and protein expression level increased gradually in each group, at the same concentration. The ratio of PI3K protein expression and p Akt/Akt protein expression in combination group was significantly higher than that in control group (P 0.05). DCI and their combination may increase the glucose consumption of Hep G2-IR cells by increasing the expression of PI3K protein and the phosphorylation of Akt. In order to achieve the purpose of improving IR, the combined effect is the most significant.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R587.1

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