التفاصيل البيبلوغرافية
| العنوان: |
METTL3 在同型半胱氨酸诱导小鼠胰岛 β 细胞自噬中的作用. (Chinese) |
| العنوان البديل: |
Role of METTL3 in homocysteine-induced autophagy in mouse islet beta cells. (English) |
| المؤلفون: |
马凌桔, 汪乐新, 迟宏扬, 张竞文, 彭红建, 高春兰, 姜怡邓, 黄 晖, 杨 力, 马胜超 2， |
| المصدر: |
Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 9/18/2024, Vol. 28 Issue 26, p4221-4225, 5p |
| الملخص (بالإنجليزية): |
BACKGROUND: Hyperhomocysteinemia is closely related to the function of islet β cells, but its specific molecular mechanism is not fully understood. OBJECTIVE: To investigate the role of N6 methyltransferase-like 3 (METTL3) in homocysteine (Hcy)-induced autophagy of mouse islet β cells. METHODS: The 3rd and 4th generation mouse islet β cells were taken for the experiment. (1) Cell modeling and grouping: cells in control group were not treated with Hcy, while those in homocysteine group were treated with 100 µmol/L Hcy for 48 hours. (2) The mouse islet β-cells were transfected with the plasmids overexpressing Ad-METTL3 and si-METTL3 according to the instructions of LipofectamineTM 2000. Three different interfering fragments were designed, and the one with the best interfering efficiency was verified and screened by PCR. (3) After transfection, the cells were divided into control group, Hcy group, Ad-NC (negative control)+Hcy group, Ad-METTL3+Hcy group, si-NC (negative control)+Hcy group and si-METTL3+Hcy group. (4) qRT-PCR and western blot were used to detect the expression levels of METTL3 and autophagy-related proteins LC3II/I and p62 in cells. Insulin level was determined by ELISA to evaluate insulin secretion capacity of islet cells. Autophagy-related proteins and insulin level were detected after overexpression and interference with METTL3. RESULTS AND CONCLUSION: Compared with the control group, the expression level of LC3II/I was increased (P < 0.05), the expression of p62 was significantly reduced (P < 0.05), and the insulin secretion capacity was significantly decreased (P < 0.05) in the Hcy group. Compared with the control group, the protein and mRNA levels of METTL3 were reduced in the Hcy group (P < 0.05). After METTL3 silencing in islet β cells, Hcy further upregulated the expression of LC3II/I (P < 0.05), significantly dowregulated the expression of p62 (P < 0.05), and increased the insulin level (P < 0.05). After overexpression of METTL3, Hcy significantly decreased the LC3II/I expression and increased the p62 expression in islet β cells (P < 0.05). To conclude, METTL3 is involved in the Hcy-induced autophagy regulation of islet β cells and plays a role in the regulation of insulin secretion. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
背景：高同型半胱氨酸(homocysteine，Hcy)血症与胰岛β细胞功能密切相关，但其具体分子机制尚不完全明确。 目的：探讨METTL3在Hcy诱导小鼠胰岛β细胞自噬中的作用。 方法：取第3，4代小鼠胰岛β细胞进行实验。①细胞模型建立和分组：对照组细胞不加入Hcy，Hcy组细胞加入浓度为100 µmol/L Hcy干预 48 h；②按LipofectamineTM 2000说明书将过表达质粒Ad-METTL3及si-METTL3转染小鼠胰岛β细胞，设计3种不同干扰片段，PCR验证、筛选出 干扰效率最好的干扰片段；③转染后实验分组：对照组、Hcy组、Ad-NC(阴性对照)+Hcy组、Ad-METTL3+Hcy组、si-NC(阴性对照)+Hcy组和 si-METTL3+Hcy组；④采用qRT-PCR及Western blot检测细胞中METTL3及细胞自噬相关蛋白LC3Ⅱ/Ⅰ、p62的表达；ELISA法测定胰岛素水平来 评价胰岛β细胞胰岛素分泌能力；分别过表达和干扰METTL3后检测细胞自噬相关蛋白及胰岛素水平。 结果与结论：①与对照组相比，Hcy组自噬相关蛋白LC3Ⅱ/Ⅰ的表达水平升高(P < 0.05)，而p62表达明显降低( P < 0.05)，胰岛素分泌能力 明显下降(P < 0.05)；②与对照组相比，Hcy组中METTL3蛋白及mRNA水平表达均降低(P < 0.05)；③胰岛β细胞中沉默METTL3后，Hcy进一 步上调了细胞中LC3Ⅱ/Ⅰ的表达(P < 0.05)，而p62表达显著下降(P < 0.05)，细胞中胰岛素水平增加(P < 0.05)；过表达METTL3后，Hcy则使 LC3Ⅱ/Ⅰ表达显著降低且p62 表达则增高( P < 0.05)；④结论：METTL3参与了Hcy诱导的胰岛β细胞自噬调控，对胰岛素的分泌发挥着调控作用。 [ABSTRACT FROM AUTHOR] |
|
Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) |
| قاعدة البيانات: |
Complementary Index |
