Department of Physiology and Key Lab of Ministry of Education in Fertility Preservation and Maintenance, Ningxia Medical University, Yinchuan, China.
Department of Physiology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Cell Mol Med. 2018 Jan;22(1):395-408. doi: 10.1111/jcmm.13328. Epub 2017 Sep 25.
Toll-like receptors (TLRs) are essential immunoreceptors involved in host defence against invading microbes. Recent studies indicate that certain TLRs activate immunological autophagy to eliminate microbes. It remains unknown whether TLRs regulate autophagy to play a role in the heart. This study examined this question. The activation of TLR3 in cultured cardiomyocytes was observed to increase protein levels of autophagic components, including LC3-II, a specific marker for autophagy induction, and p62/SQSTM1, an autophagy receptor normally degraded in the final step of autophagy. The results of transfection with a tandem mRFP-GFP-LC3 adenovirus and use of an autophagic flux inhibitor chloroquine both suggested that TLR3 in cardiomyocytes promotes autophagy induction without affecting autophagic flux. Gene-knockdown experiments showed that the TRIF-dependent pathway mediated the autophagic effect of TLR3. In the mouse model of chronic myocardial infarction, persistent autophagy was observed, concomitant with up-regulated TLR3 expression and increased TLR3-Trif signalling. Germline knockout (KO) of TLR3 inhibited autophagy, reduced infarct size, attenuated heart failure and improved survival. These protective effects were abolished by in vivo administration of an autophagy inducer rapamycin. Similar to the results obtained in cultured cardiomyocytes, TLR3-KO did not prevent autophagic flux in mouse heart. Additionally, this study failed to detect the involvement of inflammation in TLR3-KO-derived protection, as wild-type and TLR3-KO hearts were comparable in inflammatory activity. It is concluded that up-regulated TLR3 expression and signalling contributes to persistent autophagy following MI, which promotes heart failure and lethality.
Toll 样受体(TLRs)是宿主防御入侵微生物的重要免疫受体。最近的研究表明,某些 TLR 通过激活免疫自噬来消除微生物。TLRs 是否调节自噬以在心脏中发挥作用尚不清楚。本研究探讨了这个问题。观察到培养的心肌细胞中 TLR3 的激活增加了自噬成分的蛋白水平,包括 LC3-II,这是自噬诱导的特异性标志物,以及 p62/SQSTM1,这是自噬受体在自噬的最后一步通常被降解。用串联 mRFP-GFP-LC3 腺病毒转染和使用自噬流抑制剂氯喹的结果均表明,心肌细胞中的 TLR3 促进自噬诱导而不影响自噬流。基因敲低实验表明,TRIF 依赖性途径介导了 TLR3 的自噬作用。在慢性心肌梗死的小鼠模型中,观察到持续的自噬,同时伴有 TLR3 表达上调和 TLR3-Trif 信号增加。TLR3 的种系敲除(KO)抑制了自噬,减少了梗死面积,减轻了心力衰竭并提高了生存率。体内给予自噬诱导剂雷帕霉素可消除这些保护作用。与在培养的心肌细胞中获得的结果类似,TLR3-KO 并没有阻止小鼠心脏中的自噬流。此外,这项研究未能检测到 TLR3-KO 衍生的保护作用中炎症的参与,因为野生型和 TLR3-KO 心脏在炎症活性方面没有差异。综上所述,TLR3 表达和信号的上调导致 MI 后持续的自噬,促进心力衰竭和致死性。