Platform for preclinical screening of neuroprotective strategies: a clinically relevant ICH model and standardized functional test battery

Roman V. Deev; Olesya V. Shcheblykina; Chao  Zhu; Darya A. Kostina; Wan Sun; Vladimir V. Gureev; Iurii K. Slepov; Nikita S. Zhunusov; Lilia V. Korokina; Oleg S. Gudyrev; Anastasia A. Nekrasova; Olga A. Osipova; Elizaveta I. Repina; Arkady V. Nesterov; Tatiana G. Pokrovskaya

doi:10.18413/rrpharmacology.12.1043

Authors

Roman V. Deev Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery https://orcid.org/0000-0001-8389-3841
Olesya V. Shcheblykina Belgorod State National Research University https://orcid.org/0000-0003-0346-9835
Chao Zhu Dezhou University
Darya A. Kostina Belgorod State National Research University https://orcid.org/0000-0002-4505-3988
Wan Sun Dezhou Universit https://orcid.org/0000-0003-0780-4768
Vladimir V. Gureev Belgorod State National Research University https://orcid.org/0000-0003-1433-1225
Iurii K. Slepov Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery; LLC SWIFTGEN https://orcid.org/0000-0003-3498-4573
Nikita S. Zhunusov Belgorod State National Research University https://orcid.org/0000-0002-1969-3615
Lilia V. Korokina Belgorod State National Research University https://orcid.org/0000-0003-1030-7041
Oleg S. Gudyrev Belgorod State National Research University https://orcid.org/0000-0003-0097-000X
Anastasia A. Nekrasova N.N. Burdenko Voronezh State Medical University https://orcid.org/0009-0008-4043-7588
Olga A. Osipova Belgorod State National Research University https://orcid.org/0000-0002-7321-6529
Elizaveta I. Repina Belgorod State National Research University https://orcid.org/0009-0000-9508-3993
Arkady V. Nesterov Belgorod State National Research University https://orcid.org/0000-0003-3822-4213
Tatiana G. Pokrovskaya Belgorod State National Research University https://orcid.org/0000-0001-6802-5368

DOI:

https://doi.org/10.18413/rrpharmacology.12.1043

Abstract

Introduction: Intracerebral hemorrhage (ICH) is among the most severe forms of stroke. Whereas traditional hemorrhagic stroke models reproduce hemorrhage within the striatum (specifically the putamen), localization of the hematoma with involvement of the internal capsule and globus pallidus may better recapitulate clinically relevant sensorimotor deficits due to more extensive basal ganglia involvement and damage to dense clusters of white matter tracts.

Materials and Methods: Male CD1 mice (3–4 months, 30–35 g) were assigned to intact control (n=15) or ICH (n=15) groups. ICH was induced by stereotaxic injection of 30 μL autologous blood into the right cerebral hemisphere (2.3 mm lateral, 0.2 mm caudal to bregma, depths 3.0 and 3.5 mm). Neurological status was assessed using the modified Garcia scale, limb placement tests, horizontal and vertical pole tests, inverted grid test, corner test, and open field test at baseline and on days 1, 3, 7, 14, 21, and 28.

Results and Discussion: The model showed high reproducibility with 33–40% acute mortality. All ICH mice developed contralateral sensorimotor-pyramidal deficits with predominant forelimb involvement. Garcia scores declined from 21 to 14.83±1.72 on day 1 with gradual but incomplete recovery by day 28 (18 [18;18.5]). Forelimb placement showed >50% acute deficit with a nonlinear recovery pattern, whereas hindlimb deficits appeared later (days 3–7) and persisted to day 21. The modified Garcia scale, forelimb placement, modified horizontal bar, and open field tests were most sensitive to functional impairment and recovery.

Conclusion: Stereotaxic autologous blood injection into the basal ganglia (putamen and globus pallidus) with involvement of the internal capsule in mice reproduces clinically relevant, reproducible sensorimotor deficits with persistent functional impairment, making this model suitable for preclinical testing of neuroprotective strategies.

Graphical Abstract

Keywords:

hemorrhagic stroke, internal capsule, ICH model, mice, Garcia scale, behavioral assessment, sensorimotor deficit

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Author Contribution

Roman V. Deev, Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery

Cand. Sci. (Medicine), Deputy Director at Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; e-mail: romdey@gmail.com; ORCID ID: https://orcid.org/0000-0001-8389-3841. Conceptualization and study design, supervision, critical revision of the manuscript for important intellectual content, final approval of the version to be published.

Olesya V. Shcheblykina, Belgorod State National Research University

Cand. Sci. (Medicine), Associate Professor of the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: sheolvi31@gmail.com; ORCID ID: https://orcid.org/0000-0003-0346-9835. Development of the experimental design and intracerebral hemorrhage modeling protocol, supervision of the experimental work, data analysis and interpretation, drafting of the manuscript and substantial revision of its content.

Chao Zhu, Dezhou University

Shandong Provincial Engineering Laboratory of Novel Pharmaceutical Excipients, Sustained and Controlled Release Preparations, Dezhou University, Dezhou, China; e-address: zhuchao830111@163.com; ORCID ID: Provided expert consultation, contributed to the interpretation of results, and reviewed the manuscript.

Darya A. Kostina, Belgorod State National Research University

Cand. Sci. (Medicine), Associate Professor of the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: daria-f13@mail.ru; ORCID ID: https://orcid.org/0000-0002-4505-3988. Primary data processing and statistical analysis, contribution to data interpretation and preparation of the Results and Discussion sections.

Wan Sun, Dezhou Universit

Provincial Engineering Laboratory of Novel Pharmaceutical Excipients, Sustained and Controlled Release Preparations, Dezhou University, Dezhou, China; e-mail: sunwananhui@163.com; ORCID ID: https://orcid.org/0000-0003-0780-4768. Provided methodological and conceptual guidance, contributed to data interpretation.

Vladimir V. Gureev, Belgorod State National Research University

Doctor Habil. of Medical Sciences, Associate Professor, Professor of the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: produmen@mail.ru; ORCID ID: https://orcid.org/0000-0003-1433-1225. Contribution to the development of the study concept and design, definition of the methodological approach to neurological function testing.

Iurii K. Slepov, Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery; LLC SWIFTGEN

Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery, LLC SWIFTGEN, Moscow, Russia; e-mail: slepovurij95@gmail.com; ORCID ID: https://orcid.org/0000-0003-3498-4573. Consulting on pharmacological and statistical study design, participation in data analysis and interpretation.

Nikita S. Zhunusov, Belgorod State National Research University

Junior Researcher at the Laboratory of Genetic Technologies and Gene Editing for Biomedicine and Veterinary, Belgorod State National Research University, Belgorod, Russia; e-mail: nzhunuson29@gmai.com; ORCID ID: https://orcid.org/0000-0002-1969-3615. Animal handling, performance of procedures for intracerebral hemorrhage modeling, participation in behavioral testing and documentation of raw data.

Lilia V. Korokina, Belgorod State National Research University

Doctor Habil. of Medical Sciences, Associate Professor, Associate Professor of the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail address: korokina@mail.ru; ORCID ID: https://orcid.org/0000-0003-1030-7041. Participation in study planning and selection of scales/tests for neurological deficit assessment, critical expert evaluation of the methodology and results.

Oleg S. Gudyrev, Belgorod State National Research University

Cand. Sci. (Medicine), Associate Professor, Associate Professor at the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: gudyrev@mail.ru; ORCID ID: https://orcid.org/0000-0003-0097-000X. Consulting on pharmacological and statistical study design, participation in data analysis and interpretation, manuscript editing.

Anastasia A. Nekrasova, N.N. Burdenko Voronezh State Medical University

6^th year student of the faculty of Medicine, N.N. Burdenko Voronezh State Medical University, Voronezh, Russia; e-mail: nasnek111@mail.ru; ORCID ID: https://orcid.org/0009-0008-4043-7588. Collection and systematization of experimental data, and preparation of illustrative material.

Olga A. Osipova, Belgorod State National Research University

Doctor Habil. of Medical Sciences, Professor, Professor of the Department of Hospital Therapy, Belgorod State National Research University, Belgorod, Russia; e-mail: osipova@bsuedu.ru; ORCID ID: https://orcid.org/0000-0002-7321-6529. Clinico-pathophysiological interpretation of the results in the context of human stroke, comparison of experimental data with clinical observations, critical revision of the Discussion section.

Elizaveta I. Repina, Belgorod State National Research University

Medical student, Belgorod State National Research University, Belgorod, Russia; e-mail: repina.2004@gmail.com; ORCID ID: https://orcid.org/0009-0000-9508-3993. Collection and systematization of experimental data, and preparation of illustrative material.

Arkady V. Nesterov, Belgorod State National Research University

Cand. Sci. (Medicine), Associate Professor at the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: nesterov_a@yandex.ru; ORCID ID: https://orcid.org/0000-0003-3822-4213. Participation in the development and refinement of the intracerebral hemorrhage modeling technique, performance of stereotaxic procedures, ensuring standardization of surgical procedures.

Tatiana G. Pokrovskaya, Belgorod State National Research University

Doctor Habil. of Medical Sciences, Professor, Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: pokrovskaia-tg@mail.ru; ORCID ID: https://orcid.org/0000-0001-6802-5368. Overall scientific coordination of the project, contribution to the formulation of the study aim and objectives, critical revision of the manuscript.