Mettl3 inhibits the apoptosis and autophagy of chondrocytes in inflammation through mediating Bcl2 stability via Ythdf1-mediated m6A modification
Mettl3 inhibits the apoptosis and autophagy of chondrocytes in inflammation through mediating Bcl2 stability via Ythdf1-mediated m6A modification
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Mettl3 通过 Ythdf1 介导的 m6A 修饰介导 Bcl2 稳定性，抑制炎症中软骨细胞的凋亡和自噬
DOI：
10.1016/j.bone.2021.116182
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发表时间：
2022-01-01
期刊：
影响因子：
4.1
通讯作者：
Wang, Jiawei
中科院分区：
文献类型：
--
作者：
He, Ying;Wang, Wei;Wang, Jiawei
N-6甲基腺苷（m(6)A）甲基化是真核信使RNA中最常见的内部修饰之一，发生在腺苷的N-6氮上。然而，m(6)A在颞下颌关节骨性关节炎（tmjoa）中的作用尚不清楚。在这里，我们研究了甲基转移酶样3 （Mettl3）在炎症软骨细胞中的功能和机制。我们发现Mettl3的表达在TMJ OA小鼠体内和体外炎症刺激下均降低。功能、损失和增益研究表明，Mettl3在体外抑制tnf - α刺激诱导的软骨细胞凋亡和自噬。Mettl3抑制剂s -腺苷型同型半胱氨酸（SAH）在体外促进炎症软骨细胞凋亡和自噬，并在体内加重碘乙酸钠（MIA）诱导的TMJ OA小鼠软骨细胞和软骨下骨的变性。机制上，采用生物信息学分析、m(6)A-RNA免疫沉淀（MeRIP）和RNA免疫沉淀（RIP）鉴定Bcl2 mRNA是Mettl3对m(6)A修饰的下游靶点。此外，研究结果表明，Ythdf1介导了Mettl3催化Bcl2 mRNA的稳定性。共免疫沉淀（Co-IP）结果显示，在TNF-a刺激诱导的软骨细胞中，Bcl2蛋白与Beclinl蛋白相互作用。综上所述，我们的研究结果表明，Mettl3通过m(6)A/Ythdf1/Bcl2信号轴抑制炎症中软骨细胞的凋亡和自噬，这为TMJ OA的治疗提供了有希望的策略。
N-6-methyladenosine (m(6)A) methylation is one of the most common internal modifications in eukaryotic messenger RNA occurring on N-6 nitrogen of adenosine. However, the roles of m(6)A in tempommandibular joint osteoarthritis (TMJ OA) are still elusive. Here, we investigate the function and mechanism of methyltransferaselike 3 (Mettl3) in chondrocytes in inflammation. We found that the expression of Mettl3 decreased both in vivo TMJ OA mice and in vitro inflammatory stimulation. Functionally, loss and gain studies illustrated that Mettl3 inhibited the apoptosis and autophagy of chondrocytes induced by TNF-alpha stimulation in vitro. Mettl3 inhibitor, S-adenosylhomocysteine (SAH) promoted the apoptosis and autophagy of chondrocytes with inflammation in vitro and aggravated the degeneration of chondrocytes and subchondral bone in monosodium iodoacetate (MIA) induced TMJ OA mice in vivo. Mechanistically, the bioinformatics analysis, m(6)A-RNA immunoprecipitation (MeRIP) and RNA immunoprecipitation (RIP) were used to identify that Bcl2 mRNA was the downstream target of Mettl3 for m(6)A modification. Furthermore, the results revealed that Yth m(6)A RNA binding protein 1 (Ythdf1) mediated the stability of Bcl2 mRNA catalyzed by Mettl3. Co-immunoprecipitation (Co-IP) showed that Bcl2 protein interacted with Beclinl protein in chondrocytes induced by TNF-a stimulation. In conclusion, our findings identify that Mettl3 inhibits the apoptosis and autophagy of chondrocytes in inflammation through m(6)A/Ythdf1/Bcl2 signal axis which provides promising therapeutic strategy for TMJ OA.