Neuroprotective Effect of Tricyclic Pyridine Alkaloids from Fusarium lateritium SSF2, against Glutamate-Induced Oxidative Stress and Apoptosis in the HT22 Hippocampal Neuronal Cell Line

Dahae Lee; Hyun Gyu Choi; Ji Hye Hwang; Sang Hee Shim; Ki Sung Kang

doi:10.3390/antiox9111115

Neuroprotective Effect of Tricyclic Pyridine Alkaloids from Fusarium lateritium SSF2, against Glutamate-Induced Oxidative Stress and Apoptosis in the HT22 Hippocampal Neuronal Cell Line

Antioxidants (Basel). 2020 Nov 11;9(11):1115. doi: 10.3390/antiox9111115.

Authors

Dahae Lee¹, Hyun Gyu Choi², Ji Hye Hwang¹, Sang Hee Shim², Ki Sung Kang¹

Affiliations

¹ College of Korean Medicine, Gachon University, Seongnam 13120, Korea.
² College of Pharmacy, Duksung Women's University, Seoul 01369, Korea.

Abstract

Excessive glutamate damages neuronal cells via the accumulation of intracellular reactive oxygen species (ROS), calcium ion (Ca²⁺) influx, depolarization of mitochondrial membrane potential, and apoptosis, which may result in the development of chronic neurodegenerative diseases. In this study, we evaluated the effects of 4,6'-anhydrooxysporidinone isolated from endophytic fungus Fusarium lateritium SSF2 on glutamate-induced cytotoxicity, accumulation of intracellular ROS, increases in superoxide anion production, Ca²⁺, depolarization of mitochondrial membrane potential, and apoptotic cell death in hippocampal HT22 cells. 2',7'-Dichlorofluorescin diacetate (H2DCFDA) staining was used to determine the intracellular reactive oxygen species concentration and dihydroethidine (DHE) staining was used to determine the superoxide radical. Expression of the nuclear factor-erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) was analyzed by Western blot. Fluo-4 staining was used to determine the intracellular Ca²⁺ levels. In order to explore mitochondrial membrane potential, tetramethylrhodamine methyl ester (TMRM) staining was used. Apoptotic cell death was evaluated using Annexin-V/propidium iodide (PI) staining and TUNEL staining. Expression of the cytochrome c release and cleaved caspase-9, -3 was analyzed by Western blot. Here, we were able to isolate 4,6'-anhydrooxysporidinone from endophytic fungus, Fusarium lateritium SSF2, which was shown to protect HT22 cells from glutamate-induced cytotoxicity, accumulation of intracellular ROS, increases in superoxide anion production, Ca²⁺, and depolarization of mitochondrial membrane potential. In addition, 4,6'-anhydrooxysporidinone enhanced the expressions of Nrf2 and HO-1. It also inhibited the apoptotic cell death through the inhibition of cytochrome c release and cleaved caspase-9, -3 in glutamate-treated HT22 cells. Therefore, our results provide ample evidence of the neuroprotective properties of 4,6'-anhydrooxysporidinone.

Keywords: Ca2+; Fusarium lateritium SSF2; HT22 cells; apoptosis; glutamate; oxidative stress.