Role of METTL3 in homocysteine-induced autophagy in mouse islet beta cells

doi:10.12307/2024.423

Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (26): 4221-4225.doi: 10.12307/2024.423

Role of METTL3 in homocysteine-induced autophagy in mouse islet beta cells

Ma Lingju1, 2, Wang Lexin2, Chi Hongyang2, 3, Zhang Jingwen2, Peng Hongjian2, 4, Gao Chunlan2, 5, Jiang Yideng2, Huang Hui1, Yang Li1, Ma Shengchao2, 3

1Department of Geriatrics and Special Need Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3School of Inspection, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 4College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, Hunan Province, China; 5The First People’s Hospital of Yinchuan, Yinchuan 750001, Ningxia Hui Autonomous Region, China

Received:2023-06-06 Accepted:2023-07-24 Online:2024-09-18 Published:2023-10-07
Contact: Ma Shengchao, MD, Associate professor, Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Yinchuan 750004, Ningxia Hui Autonomous Region, China; School of Inspection, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
About author:Ma Lingju, Master, Physician, Department of Geriatrics and Special Need Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Supported by:
the National Natural Science Foundation of China (Youth Project), No. 81900273 (to MSC); National Natural Science Foundation of China (Regional Project), Nos. 82060139 and 82270492 (to MSC); Outstanding Youth Project of Natural Science Foundation of Ningxia Autonomous Region, No. 2023AAC05035 (to MSC); the Key Research and Development Program of Ningxia Hui Autonomous Region, No. 2019BEG03006 (to ZJW); The School-level Key Project of Ningxia Medical University, No. XZ2022004 (to PHJ)

Abstract

Abstract: 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.

Key words: methyltransferase-like 3, islet β cell, homocysteine, autophagy, LC3II/I

CLC Number:

Ma Lingju, Wang Lexin, Chi Hongyang, Zhang Jingwen, Peng Hongjian, Gao Chunlan, Jiang Yideng, Huang Hui, Yang Li, Ma Shengchao. Role of METTL3 in homocysteine-induced autophagy in mouse islet beta cells[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(26): 4221-4225.

Figures/Tables 5

References

[1] GUIEU R, RUF J, MOTTOLA G. Hyperhomocysteinemia and cardiovascular diseases. Ann Biol Clin (Paris). 2022;80(1):7-14.
[2] CHENG Y, WANG C, ZHANG X, et al. Circulating homocysteine and folate concentrations and risk of type 2 diabetes: A retrospective observational study in Chinese adults and a Mendelian randomization analysis. Front Cardiovasc Med. 2022;9:978-998.
[3] 陈辛玲,王生兰.细胞自噬过程、通路、调控及其与肺动脉高压的多重相关性[J].中国组织工程研究,2021,25(2):311-316.
[4] 丁宇,王彦丰,李小溪,等.细胞自噬与常见老年病的关联性研究进展[J].国际老年医学杂志,2022,43(4):468-471.
[5] 高源,揭育祯,马天龙,等.miR-488-3p靶向调控RAP1A在同型半胱氨酸介导肝细胞自噬的作用研究[J].中国比较医学杂志,2022,32(9):1-9.
[6] 赵钎汐,杨淑德,郭澍.METTL3对细胞分化调控的研究进展[J].中国美容整形外科杂志,2022,33(8):508-509,514-515.
[7] LI X, JIANG Y, SUN X, et al. METTL3 is required for maintaining β-cell function. Metabolism. 2021;116:154702.
[8] 张晴,高春兰,于飞飞,等.同型半胱氨酸致胰岛β细胞凋亡中肝配蛋白A型受体2 DNA甲基化升高[J].中国组织工程研究,2023,27(5):714-719.
[9] SUN H, SAEEDI P, KARURANGA S, et al. IDF Diabetes Atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045. Diabetes Res Clin Pract. 2022;183:109119.
[10] KOTWAS A, KARAKIEWICZ B, ZABIELSKA P, et al. Epidemiological factors for type 2 diabetes mellitus: evidence from the Global Burden of Disease. Arch Public Health. 2021;79(1):110.
[11] 吕承安,王若然,孟卓贤.2型糖尿病进程中胰岛β细胞功能变化的分子机制[J].遗传,2022,44(10):840-852.
[12] CASSIDY FC, SHORTISS C, MURPHY CG, et al. Impact of type 2 diabetes mellitus on human bone marrow stromal cell number and phenotypic characteristics. Int J Mol Sci. 2020;21:2476.
[13] 陈艳,王萍.同型半胱氨酸和总胆红素对胰岛β细胞功能的交互作用分析[J].中国糖尿病杂志,2023,31(1):47-52.
[14] 揭育祯,杨慧霞,周瑜瑾,等.TRPC6 DNA甲基化在同型半胱氨酸致胰岛β细胞焦亡中的作用研究[J].广东医学,2022,43(2):162-168.
[15] MIZUSHIMA N, LEVINE B. Autophagy in Human Diseases. N Engl J Med. 2020;383(16): 1564-1576.
[16] GE X, WANG L, FEI A, et al. Research progress on the relationship between autophagy and chronic complications of diabetes. Front Physiol. 2022;13:956344.
[17] KIM J, PARK K, KIM MJ, et al. An autophagy enhancer ameliorates diabetes of human IAPP-transgenic mice through clearance of amyloidogenic oligomer. Nat Commun. 2021;12(1):183.
[18] 任海文,王林华,龚权,等.胰岛β细胞自噬及其影响因素[J].中国糖尿病杂志, 2022,30(10):796-800.
[19] ZHANG Y, ZHOU XA, LIU C, et al. Vitamin B6 Inhibits High Glucose-Induced IsletβCell Apoptosis by Upregulating Autophagy. Metabolites. 2022;12(11):1048.
[20] CAI XS, SHE MQ, XU MY, et al. GLP - 1 treatment protects endothelial cells from oxidative stress -induced autophagy and endothelial dysfunction. Int J Biol Sci. 2018;14: 1696-1708.
[21] LAMBELET M, TERRA LF, FUKAYA M, et al. Dysfunctional autophagy following exposure to pro - inflammatory cytokines contributes to pancreatic β-cell apoptosis. Cell Death Dis. 2018;9(2):96.
[22] MARIÑO G, NISO SM, BAEHRECKE EH, et al. Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol. 2014;15(2):81-94.
[23] SU X, QU Y, MU D. The Regulatory Network of METTL3 in the Nervous System: Diagnostic Biomarkers and Therapeutic Targets. Biomolecules. 2023;13(4):664.
[24] WANG JN, WANG F, KE J, et al. Inhibition of METTL3 attenuates renal injury and inflammation by alleviating TAB3 m6A modifications via IGF2BP2-dependent mechanisms. Sci Transl Med. 2022;14(640):eabk2709.
[25] LI X, YUAN B, LU M, et al. The methyltransferase METTL3 negatively regulates nonalcoholic steatohepatitis (NASH) progression. Nat Commun. 2021;12(1):7213.
[26] CHEN H, PAN Y, ZHOU Q, et al. METTL3 Inhibits Antitumor Immunity by Targeting m 6 A-BHLHE41-CXCL1/CXCR2 Axis to Promote Colorectal Cancer. Gastroenterology. 2022;163(4):891-907.
[27] PENG Z, GONG Y, WANG X, et al. METTL3-m 6 A-Rubicon axis inhibits autophagy in nonalcoholic fatty liver disease. Mol Ther. 2022;30(2):932-946.
[28] 伍义文,曹健斌,黄维佳,等.m6A甲基转移酶METTL3介导miR-127调控非小细胞肺癌细胞系自噬的机制研究[J].现代检验医学杂志,2022,37(2):12-16+32.
[29] ZHOU S,SUN Y,XING Y, et al. Exenatide ameliorates hydrogen peroxide-induced pancreaticβ-cell apoptosis through regulation of METTL3-mediated m 6 A methylation. Eur J Pharmacol. 2022;924:174960.
[30] WANG Y, SUN J, LIN Z, et al. m6A mRNA Methylaon Controls Funconal Maturaon in Neonatal Murine β-Cells. Diabetes. 2020;69(8):1708-1722.
[31] CHENG Y, YAO XM, ZHOU SM, et al. The m 6 A Methyltransferase METTL3 Ameliorates Methylglyoxal-Induced Impairment of Insulin Secretion in PancreaticβCells by Regulating MafA Expression. Front Endocrinol (Lausanne). 2022;13:910868.
[32] FANG J, CHEN Z, LAI X, et al. Mesenchymal stem cells-derived HIF-1α-overexpressed extracellular vesicles ameliorate hypoxia-induced pancreaticβcell apoptosis and senescence through activating YTHDF1-mediated protective autophagy. Bioorg Chem. 2022;129:106194.

Role of METTL3 in homocysteine-induced autophagy in mouse islet beta cells

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