Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

MDPI full text link MDPI Free PMC article
Full text links

Actions

Share

.2022 Aug 29;11(9):1697.
doi: 10.3390/antiox11091697.

Olanzapine Ameliorates Ischemic Stroke-like Pathology in Gerbils and H2O2-Induced Neurotoxicity in SH-SY5Y Cells via Inhibiting the MAPK Signaling Pathway

Affiliations

Olanzapine Ameliorates Ischemic Stroke-like Pathology in Gerbils and H2O2-Induced Neurotoxicity in SH-SY5Y Cells via Inhibiting the MAPK Signaling Pathway

Md Sadikul Islam et al. Antioxidants (Basel)..

Abstract

Olanzapine (OLNZ) is used to treat psychotic disorders. To look into the neurological basis of this phenomenon, we investigated the neuroprotective effects of OLNZ in gerbils and SH-SY5Y cells. Gerbils were subjected to transient global cerebral ischemia (TGCI) by blocking both common carotid arteries, and OLNZ (10 mg/kg) was injected intraperitoneally. Hydrogen peroxide (H2O2) was used to induce oxidative-stress-mediated damage in the SH-SY5Y cells. The results indicated that OLNZ administration markedly reduced neuron damage and glial cell triggering within CA1 zone of the hippocampus. We used RNA sequencing to assess the numbers of up-and downregulated genes involved in TGCI. We found that OLNZ treatment downregulated the expression of complement-component-related genes and the expression of mitogen-activated protein kinases (MAPKs) in the hippocampus. In cells, OLNZ co-treatment significantly improved cell viability and reduced lactate dehydrogenase (LDH), and reactive oxygen species (ROS) generation. Expression of antioxidant superoxide dismutase-1,2 enzymes (SOD-1, SOD-2) was also intensely upregulated by OLNZ, while the expression of MAPKs and NF-κB were reduced. Co-incubation with OLNZ also regulated apoptosis-related proteins Bax/Bcl-2 expression. Finally, the results demonstrated that treatment with OLNZ showed neuroprotective effects and that the MAPK pathway could involve in the protective effects.

Keywords: MAPK; antioxidant; differentially expressed genes (DEGs); neuroprotection; olanzapine; oxidative stress; transient ischemia.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no competing interest.

Figures

Figure 1
Figure 1
Chemical structure of olanzapine.
Figure 2
Figure 2
Role of OLNZ in CV positive neuronal cells and F-J B histofluorescence neuronal cells in the hippocampus of CA1 domain. Identification of CV positive cells in the sham group (A) 10× and (a) 200×, TI-mediated ischemic stroke group (B) 10× and (b) 200×, and TI-mediated ischemic stroke + OLNZ (10 mg/kg) group (C) 10× and (c) 200×. F-J B histofluorescent of the CA1 domain in the sham group (D) 10× and (d) 200×, TI-mediated ischemic stroke group (E) 10× and (e) 200×, and TI-mediated ischemic stroke + OLNZ (10 mg/kg) group (F) 10× and (f) 200×. In the TI-mediated stroke gerbils, the number of CV positive cells in the CA1 domain was reduced compared with the sham group, although a greater number of F-J B+ degenerative cells were distinguished in the hippocampus of the ischemic group compared to both the sham and TI-induced stroke + OLNZ treatment groups. (G) Graphs represent the relative numbers of CV+ and (H) F-J B+ cells. Data were expressed as the mean ± SEM,n = 5 in each group. After performing one-way ANOVA with Tukey’s post hoc tests,p < 0.05 for comparisons of the treatment groups with the sham group (#) andp < 0.05 for comparisons with the TI-induced stroke group (*) were considered significant.
Figure 3
Figure 3
Role of OLNZ in NeuN immunoreactive cells in the CA1 area of the hippocampus. Identification of NeuN+ cells in the sham group (A) 10× and (a) 200x, TI-mediated ischemic stroke group (B) 10× and (b) 200x, and TI-mediated ischemic stroke + OLNZ (10mg/kg) group (C) 10× and (c) 200x. A small quantity of NeuN immunoreactive cells was superficially noticeable in the TI-mediated stroke animals in comparison with the OLNZ-treated animals. (D) The graph signifies the relative numbers of immunoreactive NeuN+ cells. Data were expressed as the mean ± SEM,n = 5 in each group. After performing one-way ANOVA with Tukey’s post hoc tests,p < 0.05 for comparisons of the treatment groups with the sham group (#) andp < 0.05 for comparisons with the TI-induced stroke group (*) were considered significant.
Figure 4
Figure 4
Role of OLNZ in Iba-1-immunoreactive microglia and GFAP-immunoreactive astrocyte cells in the CA1 area of the hippocampus. Identification of Iba-1+ neuronal cells in the sham (A) 10× and (a) 200×, TI-mediated ischemic stroke (B) 10× and (b) 200×, and TI- mediated ischemic stroke +OLNZ (10 mg/kg) (C) 10× and (c) 200× groups and of GFAP immunoreactive astrocytes in the CA1 region in the sham (D) 10× and (d) 200×, TI- mediated ischemic stroke (E) 10× and (e) 200×, and TI- mediated ischemic stroke +OLNZ (10 mg/kg) (F) 10× and (f) 200× groups. In the TI- mediated stroke group, Iba-1+ cells were notably augmented and GFAP+ cells exhibited bigger and denser processes in the CA1 area compared with OLNZ treatment groups. (G) Graphs signify the relative numbers of Iba1+ and (H) GFAP immunoreactive cells. Data were expressed as the mean ± SEM,n = 5 in each group. After performing a one-way ANOVA with Tukey’s post hoc test,p < 0.05 for comparisons of treatment groups with the sham group (#) andp < 0.05 for comparisons with the TI-induced stroke group (*) were considered significant.
Figure 5
Figure 5
Role of OLNZ in 4-HNE immunoreactive cells in the CA1 area of the hippocampus. Identification of 4-HNE cells in the sham group (A) 10× and (a) 200×, TI-mediated ischemic stroke group (B) 10× and (b) 200×, and TI-mediated ischemic stroke + OLNZ (10 mg/kg) group (C) 10× and (c) 200×. (D) The graph signifies the relative numbers of immunoreactive 4-HNE cells. Effects of OLNZ on antioxidant enzyme (SOD-2) protein expression in hippocampal tissue. The expression of (%) (E) SOD-2 proteins was increased after treatment with OLNZ. Data were expressed as the mean ± SEM,n = 3 in each group. After performing one-way ANOVA with Tukey’s post hoc tests,p < 0.05 for comparisons of the treatment groups with the sham group (#) andp < 0.05 for comparisons with the TI-induced stroke group (*) were considered significant.
Figure 6
Figure 6
Protective role of OLNZ against the TI-mediated phosphorylation of ERK, JNK, p38 protein in the brain. The expression (% of sham) of (A) pERK, (B) pJNK proteins decreased significantly, and (C) pP38 proteins also decreased but not significantly after treatment with OLNZ in TI-induced gerbil hippocampus. Data were expressed as the mean ± SEM,n = 3/group. After performing a one-way ANOVA with Tukey’s post hoc test,p < 0.05 for comparisons of treatment groups with the sham group (#) andp < 0.05 for comparisons with the stroke group (*) were considered significant.
Figure 7
Figure 7
Alterations of transcriptomic gene expression in the stroke-induced hippocampus. (A) A total of 714 DEGs was upregulated and 218 downregulated after induction of transient ischemia (TI) using RNA sequencing (Log 2 FC); (B) In comparison, 245 DEGs were downregulated while 521 were upregulated in the TI-induced stroke + OLNZ treatment group; (C) DEGs separated (MA plot) in 1-fold change (FC) compared with the sham-TI induced group; (D) TI induced stroke + OLNZ treatment group; and (E,F) A volcano plot of DEGs.
Figure 8
Figure 8
Analysis of KEGG pathways related to TI: (A) Functional pathways were downregulated by OLNZ treatment after TI; and (B) Upregulated by OLNZ treatment after TI.
Figure 9
Figure 9
Protective role of OLNZ on gene expression of complement components in the hippocampus during transient-ischemia (TI)-induced stroke in gerbils. In TI-induced stroke, gene expression of C1q, C2, C3, C4a, and C9, was significantly upregulated, whereas treatment with OLNZ markedly downregulated the expression of these genes. Data were expressed as the mean ± SEM,n = 3 in each group. After performing a one-way ANOVA with Tukey’s post hoc test,p < 0.05 for comparisons of treatment groups with the sham group (#) andp < 0.05 for comparisons with the stroke group (*) were considered significant.
Figure 10
Figure 10
Effects of OLNZ in H2O2-induced neuronal cell death in vitro: (A) The non-toxic concentration of OLNZ in SH-SY5Y cells was estimated using the MTT assay; (B) Cell viability in H2O2-mediated toxicity was also evaluated using the MTT assay; (C) The release of cytotoxic marker LDH (%); and (D) the oxidative stress marker ROS was examined. Data were expressed as the mean ± SEM, and the experiment was repeated three times. # and * indicate a statistically significant difference compared with the control and H2O2 treatment alone, respectively,p < 0.05. the oxidative stress marker.
Figure 11
Figure 11
Effects of OLNZ on antioxidant enzyme (SOD-1, SOD-2) gene and protein expression in H2O2-induced SH-SY5Y cells. The expression of (%). (A) SOD-1 and (B) SOD-2 genes and (C) SOD-1 and (D) SOD-2 proteins was increased after pre-incubation with OLNZ in a concentration-dependent manner. Data were expressed as the mean ± SEM, and the experiment was repeated three times. (#) compared to the control and (*) compared to H2O2,p < 0.05.
Figure 12
Figure 12
Protective role of OLNZ against the H2O2-mediated phosphorylation of ERK, JNK, p38, and NF-KB protein in SH-SY5Y cells. The expression (% of (A) pP38, (B) pERK, (C) pJNK, and (D) pNF-KB proteins decreased after pretreatment with OLNZ in H2O2-induced SH-SY5Y cells. Data were presented as the mean ± SEM, and the experiment was repeated three times. (#) compared to the control and (*) compared to H2O2,p < 0.05.
Figure 13
Figure 13
Protective role of OLNZ in H2O2-mediated apoptosis and on Bax and Bcl proteins in SH-SY5Y cells. The expression (%) of (A) Bax protein decreased and (B) Bcl-2 increased after pretreatment of SH-SY5Y cells with OLNZ in comparison with H2O2-mediated neurotoxicity in SH-SY5Y cells. All data are presented as the mean ± SEM, and the experiment was repeated three times. (#) compared to the control and (*) compared to H2O2,p < 0.05.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Hsieh C.H., Lin Y.J., Chen W.L., Huang Y.C., Chang C.W., Cheng F.C., Liu R.S., Shyu W.C. HIF-1α triggers long-lasting glutamate excitotoxicity via system x(c)(-) in cerebral ischaemia-reperfusion. J. Pathol. 2017;241:337–349. doi: 10.1002/path.4838. - DOI - PubMed
    1. Lipton P. Ischemic cell death in brain neurons. Physiol. Rev. 1999;79:1431–1568. doi: 10.1152/physrev.1999.79.4.1431. - DOI - PubMed
    1. Sun M.S., Jin H., Sun X., Huang S., Zhang F.L., Guo Z.N., Yang Y. Free Radical Damage in Ischemia-Reperfusion Injury: An Obstacle in Acute Ischemic Stroke after Revascularization Therapy. Oxid. Med. Cell. Longev. 2018;2018:3804979. doi: 10.1155/2018/3804979. - DOI - PMC - PubMed
    1. Lo E.H., Dalkara T., Moskowitz M.A. Mechanisms, challenges and opportunities in stroke. Nat. Rev. Neurosci. 2003;4:399–415. doi: 10.1038/nrn1106. - DOI - PubMed
    1. Zhou Y., Men L., Sun Y., Wei M., Fan X. Pharmacodynamic effects and molecular mechanisms of lignans from Schisandra chinensis Turcz. (Baill.), a current review. Eur. J. Pharmacol. 2021;892:173796. doi: 10.1016/j.ejphar.2020.173796. - DOI - PubMed

Related information

Grants and funding

LinkOut - more resources

Full text links
MDPI full text link MDPI Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp