The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration
Introduction: The 1-methyl 4-phenyl 1,2,3,6-tetrahydropiridium (MPTP) induced model of neurodegeneration in Parkinson's disease (PD) is one of the most commonly used experimental models. This neurotoxic agent , or rather its metabolite MPP+, leads to inhibition of mitochondrial complex I, an in...
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| Format: | Article |
| Language: | English |
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University of Belgrade, Medical Faculty
2021-01-01
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| Series: | Medicinski Podmladak |
| Subjects: | |
| Online Access: | https://scindeks-clanci.ceon.rs/data/pdf/0369-1527/2021/0369-15272104059B.pdf |
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| author | Blagojević Sanja Jeremić Marija Jovanović-Tucović Maja |
| author_facet | Blagojević Sanja Jeremić Marija Jovanović-Tucović Maja |
| author_sort | Blagojević Sanja |
| collection | DOAJ |
| description | Introduction: The 1-methyl 4-phenyl 1,2,3,6-tetrahydropiridium (MPTP) induced model of neurodegeneration in Parkinson's disease (PD) is one of the most commonly used experimental models. This neurotoxic agent , or rather its metabolite MPP+, leads to inhibition of mitochondrial complex I, an increase in free radicals' production and ATP depletion, all resulting in cellular demise and death. Rho-kinase is an enzyme involved with numerous cellregulatory mechanisms, such as cytoskeleton organization, axonogenesis, vesicular transport regulation and apoptosis regulation, which are all important for cell survival. Aim: Our aim was to investigate the effects of Rho-kinase inhibition on the MPP+ induced model of neurodegeneration and the role of Akt and adenosine monophosphate-activated protein kinase (AMPK) signaling pathways in this process. Material and methods: The experiments were performed on the human neuroblastoma SHSY5Y cell line. The MTT test was used to measure the viability of the cells after the MPP+ and/ or Rho-kinase inhibitor, fasudil, treatments. Changes in activation levels, or expression of pAMPK, pAkt, AMPK and Akt, were measured using the immunoblotting method, and the protein levels were quantified by densitometry. Results: The MPP+ caused a dose-dependent decrease in cellular viability, compared to the control group (untreated cells), while fasudil treatment, prior to MPP+ exposure, improved cell viability in a dose dependant manner, compared to MPP+ treatment. Analysis of activation status of target proteins showed an increase in Akt activation after the fasudil treatment, while the AMPK activation was not significantly changed. Conclusion: Inhibition of Rho-kinase using fasudil causes a decrease in MPP+ induced cell death, which is possibly mediated by an activation of the Akt/PI3K signaling pathway. |
| first_indexed | 2024-04-13T17:24:47Z |
| format | Article |
| id | doaj.art-260460f4ac2940b99db42438f63a56a7 |
| institution | Directory Open Access Journal |
| issn | 0369-1527 2466-5525 |
| language | English |
| last_indexed | 2024-04-13T17:24:47Z |
| publishDate | 2021-01-01 |
| publisher | University of Belgrade, Medical Faculty |
| record_format | Article |
| series | Medicinski Podmladak |
| spelling | doaj.art-260460f4ac2940b99db42438f63a56a72022-12-22T02:37:49ZengUniversity of Belgrade, Medical FacultyMedicinski Podmladak0369-15272466-55252021-01-01724596410.5937/mp72-335320369-15272104059BThe neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegenerationBlagojević Sanja0Jeremić Marija1https://orcid.org/0000-0002-5748-4005Jovanović-Tucović Maja2https://orcid.org/0000-0003-3959-7628Univerzitet u Beogradu, Medicinski fakultet, SerbiaUniverzitet u Beogradu, Medicinski fakultet, SerbiaUniverzitet u Beogradu, Medicinski fakultet, SerbiaIntroduction: The 1-methyl 4-phenyl 1,2,3,6-tetrahydropiridium (MPTP) induced model of neurodegeneration in Parkinson's disease (PD) is one of the most commonly used experimental models. This neurotoxic agent , or rather its metabolite MPP+, leads to inhibition of mitochondrial complex I, an increase in free radicals' production and ATP depletion, all resulting in cellular demise and death. Rho-kinase is an enzyme involved with numerous cellregulatory mechanisms, such as cytoskeleton organization, axonogenesis, vesicular transport regulation and apoptosis regulation, which are all important for cell survival. Aim: Our aim was to investigate the effects of Rho-kinase inhibition on the MPP+ induced model of neurodegeneration and the role of Akt and adenosine monophosphate-activated protein kinase (AMPK) signaling pathways in this process. Material and methods: The experiments were performed on the human neuroblastoma SHSY5Y cell line. The MTT test was used to measure the viability of the cells after the MPP+ and/ or Rho-kinase inhibitor, fasudil, treatments. Changes in activation levels, or expression of pAMPK, pAkt, AMPK and Akt, were measured using the immunoblotting method, and the protein levels were quantified by densitometry. Results: The MPP+ caused a dose-dependent decrease in cellular viability, compared to the control group (untreated cells), while fasudil treatment, prior to MPP+ exposure, improved cell viability in a dose dependant manner, compared to MPP+ treatment. Analysis of activation status of target proteins showed an increase in Akt activation after the fasudil treatment, while the AMPK activation was not significantly changed. Conclusion: Inhibition of Rho-kinase using fasudil causes a decrease in MPP+ induced cell death, which is possibly mediated by an activation of the Akt/PI3K signaling pathway.https://scindeks-clanci.ceon.rs/data/pdf/0369-1527/2021/0369-15272104059B.pdfaktampkfasudilneurodegenerationmpp+rho-kinase |
| spellingShingle | Blagojević Sanja Jeremić Marija Jovanović-Tucović Maja The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration Medicinski Podmladak akt ampk fasudil neurodegeneration mpp+ rho-kinase |
| title | The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration |
| title_full | The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration |
| title_fullStr | The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration |
| title_full_unstemmed | The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration |
| title_short | The neuroprotective effect of Rho-kinase Inhibition in 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of neurodegeneration |
| title_sort | neuroprotective effect of rho kinase inhibition in 1 methyl 4 phenylpyridinium mpp induced cellular model of neurodegeneration |
| topic | akt ampk fasudil neurodegeneration mpp+ rho-kinase |
| url | https://scindeks-clanci.ceon.rs/data/pdf/0369-1527/2021/0369-15272104059B.pdf |
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