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Published on: January 7, 2019
Mesenchymal stem cells-conditioned medium protects PC12 cells against 2,5-hexanedione-induced apoptosis via
Shuang-Yue Li1, Yuan Qi1, Shu-Hai Hu2
11 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
Abstract:
Studies suggested that the conditioned medium of mesenchymal stem cells (MSC-CM) inhibited the increased apoptosis in various cells. However, there are no reports underlying the protection of MSC-CM against 2,5-hexanedione (HD)-induced apoptosis in neural cells. In the present study, the viability was observed in PC12 cells that received HD alone or with MSC-CM by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was estimated by Hoechst 33342 staining and flow cytometry. Mitochondrial transmembrane potential was examined by rhodamine 123. Moreover, we investigated the expression of Bax and Bcl-2, cytochrome c translocation, and caspase 3 activity by real-time polymerase chain reaction, Western blot, and immunochemistry. Nerve growth factor (NGF) was examined in MSCs and MSC-CM. Our results showed that MSC-CM promoted cell survival and reduced apoptosis in HD-exposed PC12 cells. Moreover, MSC-CM significantly reversed disturbance of Bax and Bcl-2, ameliorated disruption of mitochondrial transmembrane potential, and reduced release of cytochrome c and activity of caspase 3 in HD-exposed PC12 cells. In the meantime, NGF was detected in MSCs and MSC-CM. These findings demonstrate that MSC-CM protects against HD-induced apoptosis in PC12 cells via inhibiting mitochondrial pathway. Our results indicate that NGF in MSC-CM may be involved in the protection of MSC-CM against HD-induced apoptosis. Our study clarifies the protection of MSC-CM on HD neurotoxicity and its underlying mechanism.
Insights
Mesenchymal stem cell conditioned medium (MSC-CM) protects neural cells from 2,5-hexanedione (HD) neurotoxicity. MSC-CM inhibits apoptosis by preserving mitochondrial function and reducing caspase activity, potentially via nerve growth factor (NGF).
Area of Science:
- Neuroscience
- Cell Biology
- Stem Cell Research
Background:
- Mesenchymal stem cell conditioned medium (MSC-CM) is known to inhibit apoptosis in various cell types.
- The protective effects of MSC-CM against 2,5-hexanedione (HD)-induced apoptosis in neural cells remain uninvestigated.
Purpose of the Study:
- To investigate the protective effects of MSC-CM against HD-induced apoptosis in PC12 neural cells.
- To elucidate the underlying mechanism of MSC-CM's neuroprotection, focusing on the mitochondrial pathway and nerve growth factor (NGF).
Main Methods:
- PC12 cell viability assessed using MTT assay.
- Apoptosis evaluated by Hoechst staining and flow cytometry.
- Mitochondrial potential, Bax/Bcl-2 expression, cytochrome c release, caspase 3 activity, and NGF levels were analyzed using various molecular and biochemical techniques.
Main Results:
- MSC-CM significantly enhanced PC12 cell survival and reduced apoptosis induced by HD exposure.
- MSC-CM treatment reversed HD-induced alterations in Bax/Bcl-2 expression, mitochondrial potential, and cytochrome c release.
- MSC-CM treatment decreased caspase 3 activity and NGF was detected in MSCs and MSC-CM.
Conclusions:
- MSC-CM effectively protects PC12 neural cells against HD-induced neurotoxicity.
- The protective mechanism involves the inhibition of the mitochondrial apoptosis pathway.
- Nerve growth factor (NGF) present in MSC-CM may play a crucial role in mediating these protective effects.
