郑绍越,黄永明,杨正鹏,苗 壮,薛东波.miRNA-383对急性胰腺炎腺泡细胞自噬的影响作用[J].,2017,17(1):66-69 |
miRNA-383对急性胰腺炎腺泡细胞自噬的影响作用 |
MiRNA-383 Regulates the Acinar Cell Autophagy in Acute Pancreatitis |
投稿时间:2016-04-28 修订日期:2016-05-17 |
DOI:10.13241/j.cnki.pmb.2017.01.016 |
中文关键词: miRNA-383 自噬 急性胰腺炎 |
英文关键词: miRNA-383 Autophagy Acute pancreatitis |
基金项目:国家自然科学基金项目(81370566) |
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中文摘要: |
摘要 目的:检测转染miRNA-383的急性胰腺炎细胞模型中自噬标志蛋白LC3及Beclin1的表达情况,研究miRNA-383对急性胰腺炎腺泡细胞自噬功能的影响作用。方法:在前期工作中利用生物信息技术筛选出在急性胰腺炎中表达下调的miRNA-383。培养AR42J细胞,转染miRNA-383类似物、抑制物以及阴性对照。使用牛磺胆酸钠盐以浓度200 μM刺激细胞20 min制造急性胰腺炎细胞模型,使用BZiPAR检测胰酶激活程度证明此造模方法成功后,以此条件在转染完成后刺激AR42J细胞制造急性胰腺炎细胞模型。牛磺胆酸钠盐以浓度200 μM刺激细胞20 min后提取细胞蛋白,利用western blot技术检测各组自噬标志蛋白Be- clin1的表达情况及LC3-Ⅱ与LC3-Ⅰ比值的变化。结果:急性胰腺炎细胞模型造模成功,western blot技术检测miRNA-383抑制物转染组AR42J细胞自噬标志蛋白Beclin1表达水平及LC3-Ⅱ/LC3-Ⅰ的值上调(P<0.05),miRNA-383类似物转染组细胞Be- clin1表达水平及LC3-Ⅱ/LC3-Ⅰ的值下调(P<0.05)。结论:miRNA-383可以降低急性胰腺炎时细胞自噬水平,下调miRNA-383的表达可以提高急性胰腺炎时细胞自噬水平。 |
英文摘要: |
ABSTRACT Objective: To detect the expression of autophagy marker protein LC3-Ⅱ and Beclin1 in the acute pancreatitis cell model and investigate the regulation effects of miRNA-383 on the acinar cell autophagy function in acute pancreatitis. Methods: Before the experiments the biological information analysis technology was used to screen the differently expressed miRNAs in the rat acute pan- creatitis models. The down-regulated miRNA, miRNA-383 was found. After culturing AR42J cells, the miRNA-383 mimics, miR- NA-383 inhibitor and negative control miRNA-383 were transfected into the cells to regulate the expression of miRNA-383 in each group. Taurocholic acid sodium salt with the concentration of 200 μM stimulated the AR42J celles for 20 minutes to manufacture the acute pancreatitis cell models. The activation of the pancreatic enzymes in each group was detected by the BZiPAR to confirm that the acute pancreatitis cell models were fit for the experiment in this study. The acute pancreatitis cell models were manufactured with the tau- rocholic acid sodium salt stimulation methods after the transfection of AR42J cells. The protein in the cells would be extracted after the AR42J cells stimulated by taurocholic acid sodium salt. Then the expression degree of autophagy marker protein Beclin1 and the ratio of LC3-Ⅱ to LC3-Ⅰ which extracted from each group were detected by western blot technology. Results: The way that using the tauro- cholic acid sodium salt to manufacture the acute pancreatitis cell models was fit for the experiment in this study. The expression of au- tophagy marker protein Beclin1 and the ratio of LC3-Ⅱ to LC3-Ⅰ were up-regulated in the AR42J cells which were transfected with miRNA-383 inhibitor(P<0.05); the expression of the autophagy marker protein Beclin1 and the ratio of LC3-Ⅱ to LC3-Ⅰ was down-reg- ulated in the AR42J cells which were transfected with miRNA-383 mimics(P<0.05). Conclusion: MiRNA-383 decreases the cell au- tophagy activation in acute pancreatitis. Down-regulating the expression of miRNA-383 increases the autophagy activation in acute pan- reatitis. |
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