HMGB‑1/RAGE signaling inhibition by dioscin attenuates hippocampal neuron damage induced by oxygen‑glucose deprivation/reperfusion

Liu,Aijun; Zhang,Wenqian; Wang,Shuwei; Wang,Yuan; Hong,Jun

doi:10.3892/etm.2020.9361

HMGB‑1/RAGE signaling inhibition by dioscin attenuates hippocampal neuron damage induced by oxygen‑glucose deprivation/reperfusion

Authors:
- Aijun Liu
- Wenqian Zhang
- Shuwei Wang
- Yuan Wang
- Jun Hong
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Affiliations: Department of Neurosurgery, Tangshan Gongren Hospital, Tangshan, Hebei 063000, P.R. China
Published online on: October 15, 2020 https://doi.org/10.3892/etm.2020.9361
Article Number: 231
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cerebral ischemia is one of the most common clinical diseases characterized by high morbidity and mortality. Neurocyte apoptosis and a cascade of inflammatory signals following cerebral ischemia‑reperfusion injury (IRI) may contribute to secondary brain damage, resulting in severe neurological damage. It has been reported that dioscin, a natural steroid saponin, exerts anti‑inflammatory properties against different diseases. The present study aimed to investigate the role of dioscin in oxygen‑glucose deprivation/reperfusion (OGD/R) induction in hippocampal cells in vitro and in vivo. For the in vitro study, hippocampal cells were collected from rat embryos of gestational age of E18. The oxygen‑glucose deprivation model in primary hippocampal neurons was used to mimic cerebral IRI in vitro. To select the optimum dioscin concentration and acting time, cell viability was evaluated by a Cell Counting Kit‑8 (CCK‑8) assay. Neurons subjected to OGD/R were treated with dioscin and the inflammatory cytokines, high mobility group box chromosomal protein 1 (HMGB‑1)/receptor for advanced glycation end products (RAGE) signaling molecules and apoptosis‑associated genes were determined. The intracellular reactive oxygen species (ROS) generation was detected. Furthermore, the effects of dioscin on the antioxidant defense mechanisms were evaluated by measuring the activity of glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) and the glutathione (GSH)/glutathione disulphide (GSSG) ratio. In addition, OGD/R‑induced cells were transfected with pcDNA3.1‑HMGB‑1 and treated with dioscin, and the neuronal cell apoptosis rate was determined using a terminal deoxynucleotidyl transferase‑mediated 2‑deoxyuridine 5‑triphosphate‑biotin nick‑end labeling (TUNEL) assay. The mRNA and protein expression levels of the inflammatory factors were measured using real‑time quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis, respectively. For the in vivo investigation, the oxidation and anti‑oxidation system in rat hippocampal tissue was evaluated by detecting the expression of the aforementioned oxidative stress‑associated proteins, 3‑NT as well as 8‑oxo‑deoxyguanosine (8‑OHdG). In the hippocampal region, the apoptotic rate was determined using a TUNEL assay. The results demonstrated that dioscin at a dose of 400 ng/ml significantly reversed the increase in the expression levels of the inflammatory factors and attenuated those of apoptotic cytokines induced by OGD/R. Additionally, dioscin notably reversed the OGD/R‑mediated activation of the HMGB‑1/RAGE signaling pathway in vitro and in vivo. Cell treatment with dioscin significantly attenuated ROS production and increased the activity of antioxidant enzymes. Additionally, increasing the expression of HMGB‑1 inhibited the protective effects of dioscin on cell apoptosis in the OGD/R‑induced neurons. Furthermore, HMGB‑1 overexpression reversed the antiapoptotic and anti‑inflammatory effects of dioscin on neurons. The results of the present study indicated that dioscin exerted anti‑inflammatory, antiapoptotic and antioxidant effects via the HMGB‑1/RAGE signaling pathway. These results suggest a novel perspective of the protective effects of dioscin as a prospective remedial factor for IRI.

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