Protective effect of hydrogen‑rich medium against high glucose‑induced apoptosis of Schwann cells in vitro

Yu,Yang; Ma,Xiaoye; Yang,Tao; Li,Bo; Xie,Keliang; Liu,Daquan; Wang,Guolin; Yu,Yonghao

doi:10.3892/mmr.2015.3874

Protective effect of hydrogen‑rich medium against high glucose‑induced apoptosis of Schwann cells in vitro

Authors:
- Yang Yu
- Xiaoye Ma
- Tao Yang
- Bo Li
- Keliang Xie
- Daquan Liu
- Guolin Wang
- Yonghao Yu
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Affiliations: Department of Anesthesiology, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China, Anesthesiology Reseach Institute of Tianjin, Tianjin 300052, P.R. China, Institute of Acute Abdominal Disease Research Center of Tianjin Integrated Chinese and Western Medicine, Tianjin 300102, P.R. China
Published online on: June 2, 2015 https://doi.org/10.3892/mmr.2015.3874
Pages: 3986-3992

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Abstract

Diabetic peripheral neuropathy (DPN) is considered to be one of the most prevalent and life threatening microvascular diabetic complications. DPN affects up to 50% of patients with diabetes mellitus and there are currently no efficacious therapeutic strategies available for its treatment. Previous studies have reported that oxidative stress and poly(ADP‑ribose) polymerase‑1 (PARP‑1) may be unifying factors for hyperglycemic injury. The aim of the present study was to investigate the protective effects of hydrogen‑rich medium (HM) on high glucose (HG)‑mediated oxidative stress, PARP‑1 activation and the apoptosis of Schwann cells (SCs) in vitro. The cells were divided into different groups, and were treated for 48 h. Cell viability and apoptosis were evaluated using Cell Counting kit‑8 and annexin V/propidium iodide assays, respectively. The concentrations of 8‑hydroxy‑2‑deoxyguanosine (8‑OHdG) and peroxynitrite (ONOO‑) were detected using an enzyme‑linked immunosorbent assay. The presence of intracellular oxygen free radicals was confirmed using flow cytometric analysis. Colorimetric assays were performed to determine the activity of caspase‑3, and western blotting was performed to detect the protein expression levels of PARP‑1, cleaved PARP‑1, PAR, apoptosis‑inducing factor (AIF), B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein. HG was found to induce severe oxidative stress and promote the caspase‑dependent and caspase‑independent apoptosis of SCs. Treatment with HM inhibited HG‑induced oxidative stress by suppressing hydroxyl and ONOO‑ production, levels of 8‑OHdG, caspase‑3 activity and apoptosis in the SCs. Furthermore, treatment with HM downregulated the HG‑induced release of PAR, the activation of PARP‑1 and nuclear translocation of AIF, and upregulated the expression of Bcl‑2 in the SCs. These results indicated that HM inhibited the HG‑induced‑oxidative stress‑associated caspase‑dependent and caspase‑independent apoptotic pathways in SCs. Therefore, HM may have potential as a treatment for DPN.

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