1. Academic Validation
  2. Tomatidine protects against ischemic neuronal injury by improving lysosomal function

Tomatidine protects against ischemic neuronal injury by improving lysosomal function

  • Eur J Pharmacol. 2020 Sep 5;882:173280. doi: 10.1016/j.ejphar.2020.173280.
Anil Ahsan 1 Yanrong Zheng 1 Shijia Ma 1 Mengru Liu 1 Ming Cao 1 Yue Li 1 Wanqing Zheng 1 Xinyu Zhou 1 Minhang Xin 1 Wei-Wei Hu 1 Zhong Chen 2 Xiangnan Zhang 3
Affiliations

Affiliations

  • 1 Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang University, Hangzhou, 310058, China.
  • 2 Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang University, Hangzhou, 310058, China. Electronic address: chenzhong@zju.edu.cn.
  • 3 Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang University, Hangzhou, 310058, China. Electronic address: xiangnan_zhang@zju.edu.cn.
Abstract

Cerebral ischemia is a severe neurological disorder with limited therapy. Autophagy refers to the intracellular degradation process via an autophagosome-lysosome pathway. Emerging studies indicated the neuroprotective effects of Autophagy against ischemic neuronal injury, suggesting the potential neuroprotection of autophagy-inducing compounds. Tomatidine is a gut microbiota-derived metabolite from unripe tomatoes. Tomatidine activates Autophagy either in mammal cells or C elegans. However, potential neuroprotection of tomatidine against ischemic neuronal injury has not been determined. In the present investigation, N2a cells and primary cultured mice cortical neurons were subjected to oxygen-glucose deprivation followed by reperfusion (OGD/R). Cell injury was determined by MTT and Lactate Dehydrogenase release. Autophagosomes and autolysosomes were visualized by transfecting mCherry-GFP-tandem fluorescent LC3. The protein levels of LC3, Cathepsin D, Cathepsin B, and transcription factor EB (TFEB) were detected by Western blot. Lysosomes were stained with LysoTracker Red and dequenched-bovine serum albumin (DQ-BSA red). Tomatidine alleviated OGD/R-induced injury in N2a cells and neurons. Interestingly, tomatidine treatment attenuated, rather than reinforced, the OGD/R-elevated LC3-II, which can be reversed by lysosome inhibitor. These results indicated enhanced lysosomal activity rather than autophagosome generation with tomatidine treatment in our models. Indeed, tomatidine increased the lysosome number, proteolytic activities, as well as the expression of Cathepsin D and Cathepsin B. In addition, tomatidine increased the expression and nucleus translocation of (TFEB). Besides, lysosomal inhibitors chloroquine and bafilomycin, but not wortmannin, abolished the protection of tomatidine. In conclusion, the present study revealed the neuroprotection of tomatidine against ischemic injury by promoting lysosomal activity, possibly with the involvement of TFEB-related mechanisms.

Keywords

Autophagy flux; Cerebral ischemia; Lysosomal activation; Neuroprotection; TFEB; Tomatidine.

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