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. 2023 Nov 22;20(1):7.
doi: 10.3892/br.2023.1695. eCollection 2024 Jan.

Prolonged use of temozolomide leads to increased anxiety and decreased content of aggrecan and chondroitin sulfate in brain tissues of aged rats

Anastasia V Strokotova  1 , Dmitry K Sokolov  1 , Olga P Molodykh  2 , Elena V Koldysheva  2 , Evgenii E Kliver  3   4 , Victor S Ushakov  1 , Maxim O Politko  1 , Nadezhda V Mikhnevich  1 , Galina M Kazanskaya  1 , Svetlana V Aidagulova  1   4 , Elvira V Grigorieva  1
Affiliations

Prolonged use of temozolomide leads to increased anxiety and decreased content of aggrecan and chondroitin sulfate in brain tissues of aged rats

Anastasia V Strokotova et al. Biomed Rep. .

Abstract

Chemotherapy with temozolomide (TMZ) is an essential part of anticancer therapy used for malignant tumors (mainly melanoma and glioblastoma); however, the long-term effects on patient health and life quality are not fully investigated. Considering that tumors often occur in elderly patients, the present study was conducted on long-term (4 months) treatment of adult Wistar rats (9 months old, n=40) with TMZ and/or dexamethasone (DXM) to investigate potential behavioral impairments or morphological and molecular changes in their brain tissues. According to the elevated plus maze test, long-term use of TMZ affected the anxiety of the adult Wistar rats, although no significant deterioration of brain morphology or cellular composition of the brain tissue was revealed. The expression levels of all studied heparan sulfate (HS) proteoglycans (HSPGs) (syndecan-1, syndecan-3, glypican-1 and HSPG2) and the majority of the studied chondroitin sulfate (CS) proteoglycans (CSPGs) (decorin, biglycan, lumican, brevican, neurocan aggrecan, versican, Cspg4/Ng2, Cspg5 and phosphacan) were not affected by TMZ/DXM, except for neurocan and aggrecan. Aggrecan was the most sensitive proteoglycan to TMZ/DXM treatment demonstrating downregulation of its mRNA and protein levels following TMZ (-10-fold), DXM (-45-fold) and TMZ-DXM (-80-fold) treatment. HS content was not affected by TMZ/DXM treatment, whereas CS content was decreased 1.5-2.5-fold in the TMZ- and DXM-treated brain tissues. Taken together, the results demonstrated that treatment of adult Wistar rats with TMZ had long-term effects on the brain tissues, such as decreased aggrecan core protein levels and CS chain content and increased anxiety of the experimental animals.

Keywords: aggrecan; anxiety; brain morphology; chondroitin sulfate; dexamethasone; heparan sulfate; proteoglycan; temozolomide.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Anxiety of rats in the elevated plus maze following TMZ and/or DXM treatment. (A) Percent of time spent in open arms. (B) Head dipping in open arms. (C) Percentage of time spent in closed arms. (D) Stretching in closed arms. (E) Percentage of time spent in the center. (F) Number of boli. *P<0.05 and **P<0.01 compared with control. All data are expressed as mean ± CI/2 (Origin 8.5 software). TMZ, temozolomide; DXM, dexamethasone.
Figure 2
Figure 2
Histological analysis of rat brain tissues in the control, TMZ, DXM and TMZ + DXM rat groups. (A) Hematoxylin and eosin staining (magnification, x100; scale bar, 50 µm). (B) Immunohistochemical analysis of GFAP and OLIG2 expression in the TMZ rat group. Quantitative analysis of the DAB signal was performed with ZENblue software (magnification, x400; scale bar, 50 µm). The bars represent the mean ± standard deviation from triplicate experiments (Origin 8.5; Kruskal-Wallis' test). Control, non-treated rat brain tissues; TMZ, temozolomide, DXM; dexamethasone; GFAP, glial fibrillary acidic protein; OLIG2, oligodendrocyte transcription factor 2; DAB, 3,3'-diaminobenzidine.
Figure 3
Figure 3
Expression of HSPGs and СSPGs core proteins in rat brain tissues of the control, TMZ-treated, DXM-treated and TMZ + DXM-treated experimental animals. Reverse transcription-quantitative PCR, expression normalized to that of GAPDH (OriginPro 8.5). The bars represent the mean ± standard deviation from triplicate experiments (OriginPro 8.5). ANOVA + Fisher's Least Significant Difference test; *P<0.05. Control, non-treated rat brain tissue; TMZ, temozolomide; DXM, dexamethasone; HSPG, heparan sulfate proteoglycan; CSPG, chondroitin sulfate proteoglycan; Sdc1, syndecan-1; Sdc3, syndecan-3; Gpc1, glypican-1; Hspg2, heparan sulfate proteoglycan 2/perlecan; Dcn, decorin; Bgn, biglycan; Lum, lumican; Bcan, brevican; Ncan, neurocan; Acan, aggrecan; Vcan, versican; Cspg4/Ng2, chondroitin sulfate proteoglycan 4; Cspg5, chondroitin sulfate proteoglycan 5; Ptprz1, proteintyrosine phosphatase receptor type Z1/phosphacan.
Figure 4
Figure 4
Immunohistochemical analysis of aggrecan, decorin and brevican content in the control and TMZ/DXM-treated experimental rats. Quantitative analysis of aggrecan, decorin and brevican content was performed with ZENblue software (magnification, x400; scale bar, 50 µm). The bars represent the mean ± standard deviation from triplicate experiments (OriginPro 8.5). ANOVA + Fisher's Least Significant Difference test; *P<0.05. Control, non-treated rat brain tissue; TMZ, temozolomide; DXM, dexamethasone; Acan, aggrecan; Dcn, decorin; Bcan, brevican; DAB, 3,3'-diaminobenzidine.
Figure 5
Figure 5
Dot-blot analysis of the total CS and HS content in rat brain tissues. Original representative dot blots and semi-quantitative analysis of the dot-blots (ImageJ version 1.52 software; National Institutes of Health). The bars represent the mean ± standard deviation from triplicate experiments (OriginPro 8.5). ANOVA + Fisher's Least Significant Difference test; *P<0.05. Control, non-treated rat brain tissue; TMZ, temozolomide; DXM, dexamethasone; CS, chondroitin sulfate; HS, heparin sulfate.
Figure 6
Figure 6
Immunohistochemical analysis of the CS content in the control and TMZ/DXM-treated experimental rats. Quantitative analysis of the DAB signal was performed with ZENblue software (magnification, x400; scale bar, 50 µm). The bars represent the mean ± standard deviation from triplicate experiments (OriginPro 8.5). ANOVA + Fisher's Least Significant Difference test; *P<0.05. Control, non-treated rat brain tissue; TMZ, temozolomide; DXM, dexamethasone; CS, chondroitin sulfate; DAB, 3,3'-diaminobenzidine.
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References

    1. Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N\ Engl J Med. 2005;352:987–996. doi: 10.1056/NEJMoa043330. - DOI - PubMed
    1. McAleavey PG, Walls GM, Chalmers AJ. Radiotherapy-drug combinations in the treatment of glioblastoma: A brief review. CNS Oncol. 2022;11(CNS86) doi: 10.2217/cns-2021-0015. - DOI - PMC - PubMed
    1. Yuen CA, Barbaro M, Haggiagi A. Newly diagnosed glioblastoma in elderly patients. Curr Oncol Rep. 2022;24:325–334. doi: 10.1007/s11912-022-01201-7. - DOI - PMC - PubMed
    1. Wilson MA, Schuchter LM. Chemotherapy for Melanoma. Cancer Treat Res. 2016;167:209–229. doi: 10.1007/978-3-319-22539-5_8. - DOI - PubMed
    1. Harries M, Malvehy J, Lebbe C, Heron L, Amelio J, Szabo Z, Schadendorf D. Treatment patterns of advanced malignant melanoma (stage III-IV)-A review of current standards in Europe. Eur J Cancer. 2016;60:179–189. doi: 10.1016/j.ejca.2016.01.011. - DOI - PubMed

Grants and funding

Funding: The present study was supported by the Ministry of Science and Higher Education of the Russian Federation (grant no. 122032200240-8), the Russian Science Foundation (grant no. 19-75-00051) and the Scholarship of Russian Federation President for young scientists (grant no. SP-4000.2022.4).