Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways

Background: Cerebral ischemic injury remains a major cause of high mortality, with limited effective treatments available. Inflammatory responses play a critical role in the pathophysiology of cerebral ischemia/reperfusion (I/R) injury. Suppressing inflammation is a key strategy for mitigating cereb...

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Main Authors: Fahimeh Ramshini, Javad Amini Mahabadi, Reza Bayat, Sayyed Alireza Talaei, Zeinab Vahidinia, Hassan Hassani Bafrani
Format: Article
Language:English
Published: Elsevier 2025-12-01
Series:IBRO Neuroscience Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667242125001149
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author Fahimeh Ramshini
Javad Amini Mahabadi
Reza Bayat
Sayyed Alireza Talaei
Zeinab Vahidinia
Hassan Hassani Bafrani
author_facet Fahimeh Ramshini
Javad Amini Mahabadi
Reza Bayat
Sayyed Alireza Talaei
Zeinab Vahidinia
Hassan Hassani Bafrani
author_sort Fahimeh Ramshini
collection DOAJ
description Background: Cerebral ischemic injury remains a major cause of high mortality, with limited effective treatments available. Inflammatory responses play a critical role in the pathophysiology of cerebral ischemia/reperfusion (I/R) injury. Suppressing inflammation is a key strategy for mitigating cerebral I/R injury, making it a promising therapeutic target for stroke. Vitamin D supplementation has been revealed to exhibit anti-inflammatory and neuroprotective properties during I/R injury; however, the underlying protective mechanisms are not yet fully understood. This study aimed to investigate the effects of post-ischemic calcitriol treatment on ischemic stroke, focusing specifically on the TLR4/MyD88/NF-κB and FGFR2 signaling pathways Methods: Male Wistar rats were divided into three main groups: sham, I/R+ Vehicle, and I/R+ Calcitriol. An experimental I/R model was created by occluding the middle cerebral artery (MCA) for 1 h, followed by a 72-h reperfusion period. Calcitriol (1 μg/kg) was administered intraperitoneally for three consecutive days post-stroke. Neurological deficit scores and infarct size were evaluated 72 h after MCAO. Gene expression levels of TLR4, MyD88, NF-κB, and FGFR2 in the brain cortex were measured using RT-PCR. Additionally, histopathological changes in the cortex were examined with Nissl staining. A molecular docking analysis was performed to investigate the interactions of calcitriol with TLR4 and FGFR2, providing insights into their binding affinities and potential functional implications. Results: Our findings indicated that calcitriol treatment significantly enhanced neurological function (P < 0.05) and reduced infarct volume (P < 0.001) in cerebral I/R injury. Furthermore, calcitriol decreased the number of damaged neurons while markedly increasing the count of neurons with normal morphology (P < 0.001). Consistent with the results from molecular docking showing that calcitriol antagonizes TLR4 and FGFR2, RT-PCR analysis also revealed that calcitriol significantly suppressed the upregulation of TLR4 (P < 0.05), MyD88 (P < 0.01), NF-κB (P < 0.01), and FGFR2 (P < 0.001) mRNA expression levels. Conclusion: The results demonstrate that calcitriol treatment offers significant neuroprotective benefits following cerebral I/R injury. These protective effects may be mediated, at least in part, by the inhibition of inflammation through the TLR4/MyD88/NF-κB and FGFR2 signaling pathways. This study enhances our understanding of the molecular mechanisms involved in calcitriol's neuroprotective actions.
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spelling doaj-art-3a8752014bca4d0088e6e2735e29176c2025-08-20T03:14:36ZengElsevierIBRO Neuroscience Reports2667-24212025-12-011934535310.1016/j.ibneur.2025.06.018Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathwaysFahimeh Ramshini0Javad Amini Mahabadi1Reza Bayat2Sayyed Alireza Talaei3Zeinab Vahidinia4Hassan Hassani Bafrani5Anatomical Sciences Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, IranGametogenesis Research Center, Kashan University of Medical Sciences, Kashan, IranDepartment of Cell and Molecular Biology, Faculty of Chemistry, University of Kashan, Kashan, IranPhysiology Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, IranAnatomical Sciences Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, Iran; Correspondence to: Department of Anatomy, School of Medicine, Kashan University of Medical Sciences, Qotb-e Ravandi Blvd, Kashan 8715988141, Iran.Anatomical Sciences Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, Iran; Correspondence to: Department of Anatomy, School of Medicine, Kashan University of Medical Sciences, Qotb-e Ravandi Blvd, Kashan 8715988141, Iran.Background: Cerebral ischemic injury remains a major cause of high mortality, with limited effective treatments available. Inflammatory responses play a critical role in the pathophysiology of cerebral ischemia/reperfusion (I/R) injury. Suppressing inflammation is a key strategy for mitigating cerebral I/R injury, making it a promising therapeutic target for stroke. Vitamin D supplementation has been revealed to exhibit anti-inflammatory and neuroprotective properties during I/R injury; however, the underlying protective mechanisms are not yet fully understood. This study aimed to investigate the effects of post-ischemic calcitriol treatment on ischemic stroke, focusing specifically on the TLR4/MyD88/NF-κB and FGFR2 signaling pathways Methods: Male Wistar rats were divided into three main groups: sham, I/R+ Vehicle, and I/R+ Calcitriol. An experimental I/R model was created by occluding the middle cerebral artery (MCA) for 1 h, followed by a 72-h reperfusion period. Calcitriol (1 μg/kg) was administered intraperitoneally for three consecutive days post-stroke. Neurological deficit scores and infarct size were evaluated 72 h after MCAO. Gene expression levels of TLR4, MyD88, NF-κB, and FGFR2 in the brain cortex were measured using RT-PCR. Additionally, histopathological changes in the cortex were examined with Nissl staining. A molecular docking analysis was performed to investigate the interactions of calcitriol with TLR4 and FGFR2, providing insights into their binding affinities and potential functional implications. Results: Our findings indicated that calcitriol treatment significantly enhanced neurological function (P < 0.05) and reduced infarct volume (P < 0.001) in cerebral I/R injury. Furthermore, calcitriol decreased the number of damaged neurons while markedly increasing the count of neurons with normal morphology (P < 0.001). Consistent with the results from molecular docking showing that calcitriol antagonizes TLR4 and FGFR2, RT-PCR analysis also revealed that calcitriol significantly suppressed the upregulation of TLR4 (P < 0.05), MyD88 (P < 0.01), NF-κB (P < 0.01), and FGFR2 (P < 0.001) mRNA expression levels. Conclusion: The results demonstrate that calcitriol treatment offers significant neuroprotective benefits following cerebral I/R injury. These protective effects may be mediated, at least in part, by the inhibition of inflammation through the TLR4/MyD88/NF-κB and FGFR2 signaling pathways. This study enhances our understanding of the molecular mechanisms involved in calcitriol's neuroprotective actions.http://www.sciencedirect.com/science/article/pii/S2667242125001149CalcitriolIschemic strokeNeuroinflammationNF-kβTLR4MyD88
spellingShingle Fahimeh Ramshini
Javad Amini Mahabadi
Reza Bayat
Sayyed Alireza Talaei
Zeinab Vahidinia
Hassan Hassani Bafrani
Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
IBRO Neuroscience Reports
Calcitriol
Ischemic stroke
Neuroinflammation
NF-kβ
TLR4
MyD88
title Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
title_full Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
title_fullStr Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
title_full_unstemmed Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
title_short Therapeutic potential of calcitriol in cerebral ischemia/reperfusion injury: In vivo and in silico insights into TLR4 and FGFR2 pathways
title_sort therapeutic potential of calcitriol in cerebral ischemia reperfusion injury in vivo and in silico insights into tlr4 and fgfr2 pathways
topic Calcitriol
Ischemic stroke
Neuroinflammation
NF-kβ
TLR4
MyD88
url http://www.sciencedirect.com/science/article/pii/S2667242125001149
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