Apocynum venetum leaf extract protects against H2O2-induced oxidative stress by increasing autophagy in PC12 cells

Feng,Yuelai; Jiang,Chang; Yang,Feng; Chen,Zixian; Li,Zheng

doi:10.3892/br.2020.1313

Apocynum venetum leaf extract protects against H2O2-induced oxidative stress by increasing autophagy in PC12 cells

Authors:
- Yuelai Feng
- Chang Jiang
- Feng Yang
- Zixian Chen
- Zheng Li
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Affiliations: Department of International Baccalaureate Diploma Program, Shanghai Pinghe School, Shanghai 200127, P.R. China, Department of Orthopaedics, Zhongshan Hospital of Fudan University, Shanghai 200032, P.R. China, Integrated Traditional and Western Medicine, Zhongshan Hospital of Fudan University, Shanghai 200032, P.R. China
Published online on: June 9, 2020 https://doi.org/10.3892/br.2020.1313
Article Number: 6
Copyright: © Feng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The effect of generated.Italic@104c035a leaf extract (AVLE) on the nervous system has been widely studied, but its effect on injured neurons is not fully understood. In the present study, the protective effect of AVLE on injured neurons was determined. H2O2 was used to induce oxidative stress in PC12 cells and cell viability assays were used to determine the optimum concentration range of AVLE and its protective effects against oxidative stress. A live-dead assay was performed to confirm the effects of AVLE on oxidative stress. Subsequently, expression of apoptotic proteins including Bax and cleaved-caspase-3 were evaluated to determine whether AVLE affected apoptosis, and reactive oxygen species (ROS) levels were detected to determine the role of AVLE in H2O2 exposure. Furthermore, expression of autophagic proteins including LC3-II and p62 were detected to evaluate the effects of AVLE on autophagic activity, and cells were treated with 3-methyladenine (3-MA), an autophagic inhibitor, to identify the underlying protective mechanism of AVLE. The results showed that the optimum conditions to induce oxidative stress were treatment with 40 µM H2O2 for 2 h, and the suitable range of AVLE concentrations was shown to be 1-100 µg/ml. AVLE improved cell viability in PC12 cells following treatment with H2O2. AVLE reduced the expression of Bax and cleaved-caspase-3, and decreased ROS production. Furthermore, AVLE upregulated LC3-II expression and downregulated p62 expression, whereas treatment with 3-MA increased the levels of ROS and apoptotic proteins. These results suggest that AVLE may protect injured neurons against oxidative stress-induced apoptosis, and this effect may be associated with the reduction of ROS by increasing autophagy.

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