本文選題：ARPE-19細(xì)胞 + 糖尿病視網(wǎng)膜疾病�。� 參考：《重慶醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的：觀察高糖對體外培養(yǎng)的ARPE-19細(xì)胞TLR2表達(dá)的影響及機制。（1）利用RealtimePCR技術(shù)探討TLR2mRNA在不同D-葡萄糖濃度下培養(yǎng)人視網(wǎng)膜色素上皮細(xì)胞（retinalpigmentepithelialcell,RPE）中的表達(dá)。（2）利用Westernblot技術(shù)探討不同D-葡萄糖濃度和PKC-α抑制劑對RPE細(xì)胞TLR2蛋白和PKC-α從細(xì)胞漿向細(xì)胞膜轉(zhuǎn)位情況的影響。（3）初步探討TLR2與糖尿病視網(wǎng)膜病變（diabetesretinopathy，DR）的相關(guān)性，高糖對ARPE-19細(xì)胞TLR2表達(dá)的影響以及機制，為相關(guān)疾病的診斷治療提供新的思路。方法：1.培養(yǎng)ARPE-19細(xì)胞。2.取第16～20代人RPE細(xì)胞用于實 驗，，各組ARPE-19（RPE細(xì)胞系）細(xì)胞分別刺激48h，正常糖濃度組（5.5mmol/LD-葡萄糖）、中度高糖組（15.5mmol/LD-葡萄糖)、高糖組（25.5mmol/LD-葡萄糖)、PKC-α抑制劑組（25.5mmol/LD-葡萄糖加10mmol/LGo6976PKC-α抑制劑）、高滲對照組（5.5mmol/LD-葡萄糖加20mmol/L甘露醇）。3.Real-TimePCR分析各組ARPE-19細(xì)胞TLR2的mRNA表達(dá)水平。4.WesternBlot檢測各組ARPE-19細(xì)胞TLR2蛋白表達(dá)水平，以及PKC-α蛋白胞漿向胞膜的轉(zhuǎn)位情況。結(jié)果：與正常糖濃度組ARPE-19細(xì)胞相比，高糖組ARPE-19細(xì)胞和 中度高糖組ARPE-19細(xì)胞TLR2的mRNA和蛋白表達(dá)，以及PKC-α從細(xì)胞漿向細(xì)胞膜的轉(zhuǎn)位均增加（P0.05），并且高糖組ARPE-19細(xì)胞高于中度高糖組ARPE-19細(xì)胞（P0.05）。與高糖組ARPE-19細(xì)胞比較，PKC-α抑制劑組ARPE-19細(xì)胞的TLR2蛋白表達(dá)降低（P0.05）,同時PKC-α蛋白從細(xì)胞膜向細(xì)胞漿轉(zhuǎn)位情況顯著降低（P0.01）。 結(jié)論：1.人ARPE-19細(xì)胞有TLR2蛋白表達(dá)。2.高糖可以上調(diào)ARPE-19細(xì)胞TLR2的表達(dá)，并且隨著糖濃度的升高TLR2表達(dá)也升高。3.PKC-α抑制劑可以部分拮抗高糖上調(diào)ARPE-19細(xì)胞TLR2表達(dá)的作用。4.高糖可能是通過增加PKC-α從細(xì)胞漿向細(xì)胞膜轉(zhuǎn)位，激活PKC-α信號通路，來上調(diào)TLR2的表達(dá)。TLR2信號通路可能參與糖尿病視網(wǎng)膜病變的發(fā)生發(fā)展過程，為研究疾病的診治提供新的思路。
[Abstract]:Objective: to observe the effect of high glucose on the expression of TLR2 in cultured ARPE-19 cells and its mechanism. (1) to investigate the expression of TLR2mRNA in cultured human retinal pigment epithelial cells (RPE) by using RealtimePCR technique. With the effect of D- glucose concentration and PKC- 偽 inhibitor on the translocation of TLR2 protein and PKC- 偽 from cytoplasm to cell membrane in RPE cells, the relationship between TLR2 and diabetic retinopathy was studied. The effect of high glucose on the expression of TLR2 in ARPE-19 cells and its mechanism provide new ideas for the diagnosis and treatment of related diseases. Method 1: 1. Culture of ARPE-19 cells. Human RPE cells of the 16th ~ (th) ~ (th) passage for solid use Test, ARPE-19(RPE cells were stimulated for 48h, normal glucose concentration group was 5.5 mmol / L LD- glucose group, moderate high glucose group was 15.5 mmol / L LD- glucose group, high glucose group was 25.5 mmol / L LD- 偽 inhibitor group and 10 mmol / L LD- 偽 PKC- 偽 inhibitor, and hypertonic control group was 5.5 mmol LD- glucose + 20mmol/L mannose. The mRNA expression level of TLR2 in ARPE-19 cells was analyzed by Real-Time PCR. 4. Western Blot was used to detect the expression of TLR2 protein in ARPE-19 cells. And the translocation of PKC- 偽 protein from cytoplasm to membrane. Results: compared with the normal glucose concentration group, the ARPE-19 cells and the ARPE-19 cells in the high glucose concentration group were higher than those in the normal glucose concentration group. The expression of mRNA and protein in ARPE-19 cells and the translocation of PKC- 偽 from cytoplasm to cell membrane increased in moderate high glucose group, and the ARPE-19 cells in high glucose group were higher than ARPE-19 cells in medium high glucose group. Compared with the high glucose group, the expression of TLR2 protein in the ARPE-19 cells decreased significantly, and the translocation of PKC- 偽 protein from the cell membrane to the cytoplasm decreased significantly. Conclusion 1. TLR2 protein was expressed in human ARPE-19 cells. High glucose could up-regulate the expression of TLR2 in ARPE-19 cells, and the expression of TLR2 increased with the increase of glucose concentration. 3. PKC- 偽 inhibitor partly antagonized the up-regulation of TLR2 expression in ARPE-19 cells by high glucose. High glucose may increase the translocation of PKC- 偽 from cytoplasm to cell membrane and activate PKC- 偽 signal pathway to up-regulate the expression of TLR2. TLR2 signaling pathway may participate in the development of diabetic retinopathy and provide a new idea for the diagnosis and treatment of diabetic retinopathy.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R774.1

【參考文獻】

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1 戴超,卞修武,史景泉;諾帝抑制糖尿病大鼠視網(wǎng)膜VEGF和iNOS的表達(dá)及意義[J];第三軍醫(yī)大學(xué)學(xué)報;2005年12期

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