Improved Rat Stroke Model by Intraluminal Middle Cerebral Artery Occlusion: A Special Emphasis on Surgical Technique

Orădan, Alex; Huţanu, Adina; Horváth, Emőke; Chiriac, Liviu; Dobreanu, Minodora

doi:10.5114/hpc.2017.70008

Cited by 5 publications

(5 citation statements)

References 12 publications

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“…To mimic all aspects of transient ischemia/reperfusion, we used a microsurgical procedure recommended by Güzel (Güzel et al, 2014), with minor modifications as described (Oradan et al, 2017), followed by a complex algorithm that includes neurological assessment, evaluation by MRI measurement of ischemic-tissue volume, and a histopathological and immunohistochemical examination. Our success rate of inducing an infarction by a 90-minute middle cerebral artery occlusion was 90 % (only one animal had no ischemic lesions according to the MRI examination), in accordance with another report (Bacigaluppi, tomi & Hermann, 2010).…”

Section: Discussionmentioning

confidence: 99%

Inflammatory cell populations in early neuroinflammation in the core and peri-ischemic lesions of rat brain after transient focal cerebral ischemia: A morphometric study

Horváth

Orădan

Chiriac

et al. 2018

Preprint

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Background and Objective:Clinical and experimental observations emphasize the role of inflammation as a direct risk factor for stroke. To better characterize the inflammation, we have conducted a detailed histological analysis of the inflammatory cell population after transient middle cerebral artery occlusion in a rat model. Methods:Fifteen adult Wistar male rats were divided randomly into test (n=10) and sham (n=5) groups. In the ischemic group, transient focal cerebral ischemia was induced with an intraluminal filament technique. Histologic lesions of the ischemic core and the surrounding penumbra zone were evaluated, based on a complex algorithm. Representative morphological changes in the core and the penumbra zone were compared. Immunohistochemistry was performed for leukocytes markers (CD15, CD68, CD3), leukocyte-released effectors (MMP-9 and COX-2), and FXIII (possibly involved in microglia and macrophage activation)Results: Neuronal vacuolation and degeneration were significantly more in the core lesion, whereas cellular edema and inflammatory infiltrate were increased in the penumbra. CD68, CD3, FXIII and Cox-2 expression were significantly higher in the penumbra than in the core (p=0.026; p=0.006; p=0.002; and p<0.001).Discussion: In the rat model of middle cerebral artery occlusion, inflammatory mechanisms, microglia/macrophage cells, and T-lymphocytes likely play an important role in the penumbra. The deterioration of neurons is less in the penumbra than in the core. Appreciation of the role of the inflammatory cells and mechanisms involved in stroke might lead to measures to inhibit the injury and save brain volume. ABSTRACT Background and Objective: Clinical and experimental observations emphasize the role of inflammation as a direct risk factor for stroke. To better characterize the inflammation, we have conducted a detailed histological analysis of the inflammatory cell population after transient middle cerebral artery occlusion in a rat model.

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Section: Discussionmentioning

confidence: 99%

Inflammatory cell populations in early neuroinflammation in the core and peri-ischemic lesions of rat brain after transient focal cerebral ischemia: A morphometric study

Horváth

Orădan

Chiriac

et al. 2018

Preprint

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“…After 21 days, in I and T groups, transient focal brain ischemia was achieved during MCAO surgery, using the intraluminal filament technique. The surgical method, anaesthesia, preoperative and postoperative monitoring conditions and the measurement of the cerebral infarction volume were performed as described in a previous study [26]. Briefly, all groups were subjected to anaesthesia (3.5% isoflurane for induction and 1.5% for maintenance) and surgical procedure in order to prepare the carotid bifurcation and to occlude the left middle cerebral artery (MCA).…”

Section: Unilateral Tmcao and Reperfusion In Ratsmentioning

confidence: 99%

Fish-Oil Preconditioning Up-Regulates Expression of Splenic Arg1 Positive M2 Type Macrophages and the Arg1/Inos2 Ratio After Experimental Induced Transient Cerebral Ischemia

Horváth¹

2019

FARMACIA

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During a stroke, the contraction of the spleen can mobilize inflammatory cells that infiltrate the damaged brain tissue. We analysed the effects of transient cerebral ischemia and fish oil preconditioning on the spleen volume, architecture, and macrophage polarization. Wistar rats were subjected to middle cerebral artery occlusion (ischemic and treatment groups, each n = 10), and were sacrificed after 24 hrs, along with a sham group (n = 10). The spleen volume suffered only mild variations, while the marginal zone showed size reduction in the ischemic group, compared to treated and control groups. Granulopoiesis and myeloperoxidase positivity were significantly increased in spleens of ischemic animals (p < 0.001 vs. the treatment and sham groups). Arginase 1 was up-regulated and the Arg1/iNOS2 expression ratio showed a significant shift on macrophages of marginal zone/red pulp interface in the treatment group compared to ischemic rats (p < 0.001). Arg1 positive M2 type monocytes/macrophages participate in resolution and tissue repair thus may feature as potential therapeutical targets in cerebral ischemia. RezumatÎn ischemia cerebrală acută, contracția splinei mobilizează celule inflamatorii care infiltrează teritoriul lezat. Am analizat efectele ischemiei cerebrale tranzitorii și a precondiționării cu ulei de pește asupra variațiilor de volum, arhitectură și polarizare a macrofagelor în splina de șobolani Wistar randomizați în grup ischemic și tratat (n = 10 fiecare), supuși ocluziei arterei cerebrale medii, sacrificați după 24 ore, împreună cu un grup control. Volumul splenic a suferit variații moderate, în timp ce zona marginală a scăzut în diametru la grupul ischemic în comparație cu cel tratat și control. Granulopoieza și expresia mieloperoxidazei a crescut semnificativ în splina animalelor cu ischemie față de cele tratate și control (p < 0,001). Expresia arginazei 1 și raportul expresiei Arg1/iNOS2 la nivelul macrofagelor din interfața zonei marginale/pulpă roșie s-a deplasat în favoarea grupului tratat față de cel ischemic (p < 0,001). Deoarece macrofagele/monocitele Arg1 pozitive participă în rezoluție și reparație tisulară, pot fi ținte terapeutice în atacul ischemic cerebral.

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“…The S group, served as control, being subjected to similar anaesthesia and surgery with no filament insertion. The surgical procedure, anaesthesia, preoperative and postoperative monitoring conditions were performed as previously reported (18). Two animals from the placebo group (P 1 and P 4 ) died shortly during the post-ischemic/reperfusion period, one animal from group P (P 3 ) had a negligible ischemic volume on MRI examination, consequently they were excluded from the histological study.…”

Section: The Experimental Protocolmentioning

confidence: 99%

N-3 polyunsaturated fatty acids induce granulopoiesis and early monocyte polarization in the bone marrow of a tMCAO rat model

Horváth

Huţanu

Orădan

et al. 2019

Revista Romana De Medicina De Laborator

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Introduction: Experimental acute cerebral ischemia quickly triggers circulating inflammatory cells, provoking infiltration of neutrophils and macrophages in the damaged brain region. N-3 polyunsaturated fatty acids alleviate the ischemic deterioration, however, their potential effect on bone marrow cell mobilization is less known. Materials and methods: healthy male Wistar rats were submitted to intraperitoneal saline injection (n=10, sham Group), transient middle cerebral artery occlusion (tMCAO) and saline injection (n=10, placebo Group), tMCAO and highly purified fish-oil administration (n=10, T Group). At the two latter groups, twenty-four hours after tMCAO, MRI scans were performed to identify the ischemic regions; the eligible animals were sacrificed, the left parietal bones being removed and subjected to qualitative and quantitative histological and immunohistochemical analysis. Results: The active hematopoietic surface was maximal at the T-Group, being significantly lower in the P- and S-Groups (p=0.006 and p= 0.017). The MPO positive surface increased significantly in the T-compared to the S-Group (22.57± 0.86 % vs. 18.87± 0.68%, p= 0.004). Arg1 expression was significantly higher (p=0.001), while iNOS expression was lower (p=0.004) in the T- than in the P-Group, but similar to the S-group. The Arg1/iNOS2 ratio was higher in the FO-treated than in the P-group (p<0.001). Conclusion: the ischemic conditions triggered granulopoiesis and the increase of iNOS2 positive, type M1 macrophage in the rat bone marrow. Fish-oil treatment generated the expansion of overall hematopoietic surface along with normalization of iNOS2, up-regulating the potentially protective Arg1 positive M2 type macrophages and causing a significant shift in the M2/M1 ratio.

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Improved Rat Stroke Model by Intraluminal Middle Cerebral Artery Occlusion: A Special Emphasis on Surgical Technique

Cited by 5 publications

References 12 publications

Inflammatory cell populations in early neuroinflammation in the core and peri-ischemic lesions of rat brain after transient focal cerebral ischemia: A morphometric study

Inflammatory cell populations in early neuroinflammation in the core and peri-ischemic lesions of rat brain after transient focal cerebral ischemia: A morphometric study

Fish-Oil Preconditioning Up-Regulates Expression of Splenic Arg1 Positive M2 Type Macrophages and the Arg1/Inos2 Ratio After Experimental Induced Transient Cerebral Ischemia

N-3 polyunsaturated fatty acids induce granulopoiesis and early monocyte polarization in the bone marrow of a tMCAO rat model

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