Whole-body vibration protects against early brain injury and neuroinflammation after experimental subarachnoid hemorrhage

doi:10.21203/rs.3.rs-2340178/v1

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Whole-body vibration protects against early brain injury and neuroinflammation after experimental subarachnoid hemorrhage

https://doi.org/10.21203/rs.3.rs-2340178/v1

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Whole body vibration (WBV), as a form of physical stimulation through mechanical vibration, has been proved to have neuroprotective effects on a variety of central nervous system (CNS) diseases. However, whether WBV plays a neuroprotective role in early brain injury (EBI) after subarachnoid hemorrhage (SAH) has not been well demonstrated. Herein, we focused on investigating the potential mechanism of the therapeutic effects of WBV on SAH-induced mice. The endovascular perforation was performed to induce SAH in C57BL/6J male mice. The mice were exposed to WBV twice a day at a frequency of 30 Hz for 20 days. The curative actions of WBV were assessed using the modified Garcia scale and the beam balance scoring system, along with measuring brain water content 24 h after SAH induction. TUNEL staining was performed to observe the apoptotic cells. Immunofluorescence staining was used to evaluate the expression of astrocytes and microglia in mice's cerebral cortex. Additionally, the ELISA assay was performed to detect inflammatory cytokines, such as IL-10, IL-18, and IL-1β. Western blot was conducted to explore the expression analysis of apoptosis-associated proteins (cleaved Caspase-3). Morris Water Maze (MWM) test and rotarod test were used to evaluate the long-term neurological function of mice. Nissl staining was used to evaluate the loss of neurons in the hippocampus of mice. Our study illustrated that WBV can reduce brain water content without significantly affecting the weight of mice. Also, the TUNEL-positive cell counts of the cerebral cortex of mice in the SAH+WBV group were significantly reduced compared with that in the SAH group. The protein level of cleaved Caspase-3 in the SAH+WBV group was also decreased than that in the SAH group. Immunofluorescence staining showed that WBV suppressed the high expression of GFAP and Iba-1 caused by SAH. MWM assay and rotarod test revealed that the long-term neurological dysfunction of mice following SAH was attenuated by WBV treatment compared with SAH-induced mice, which may be closely related to the low level of neuronal loss in the hippocampal regions. Our research demonstrated that WBV treatment can reduce EBI and neuroinflammation and improve the long-term neurological dysfunction of mice after SAH, which provides a new possibility for clinical treatment of SAH in the future.

As a lethal disease of the cerebrovascular system that features extremely high mortality and disability, subarachnoid hemorrhage (SAH) is mostly attributed to intracranial aneurysm rupture [1]. Evidence has indicated that a major prognostic factor in SAH patients is early brain injury (EBI), which occurs within 72 h of SAH onset [2]. Therefore, one of the keys to improving the prognosis of patients with SAH may be to reduce EBI.

More and more evidence shows that whole body vibration (WBV), as a method of rehabilitation exercise through the vibration of a specific frequency, plays an important role in a variety of diseases, such as cerebral palsy, multiple sclerosis, Parkinson's disease (PD), and stroke [3]. Oroszi et al. verified that the cognitive and motor impairment caused by aging may be significantly improved by the treatment of WBV [4]. In addition, the inflammatory markers and infarct volume in rat tissue can be obviously reduced, while the brain-derived neurotrophic factor can be increased by the treatment of WBV [5]. Chen et al. confirmed that WBV treatment can attenuate neuroinflammation and neuronal apoptosis caused by traumatic brain injury (TBI) [6]. However, whether WBV plays a neuroprotective role in SAH remains obscure. Therefore, the current study is to investigate the underlying mechanism of the neuroprotective effects of WBV in SAH.

2.1 Experimental animals

The present experimental design was approved by the Animal Ethics Committee of the Shandong First Medical University in China, which also conformed to the animal protection law stipulations. C57BL/6J male mice were procured from the laboratory of Shandong First Medical University. These mice were raised in 25°C animal houses and had free access to food and water.

2.2 Experimental design

2.2.1 Experiment 1

To explore the neuroprotective effects of WBV, 26 C57BL/6J male mice were randomly divided into 3 groups: (1) Sham group (n = 6); (2) SAH group (n = 12, 4 mice were excluded for death and 2 mice were excluded due to the SAH grade score lower than 8); (3) SAH + WBV group (n = 8, 2 mice were excluded for death). During the 20 days of WBV treatment and subsequent experiments process, we measured the weight of the experimental mice once every 5 days to ensure that there was no statistical difference in the weight of mice in each group. Firstly, the modified Garcia scale and beam balance score system were performed to evaluate the neurological function and motor function. Finally, the wet-dry method was used to measure the brain water content of mice.

2.2.2 Experiment 2

To investigate the neuroprotective effects of WBV on neuroinflammation and neuronal apoptosis, 24 C57BL/6J male mice were randomly divided into 3 groups: (1) Sham group (n = 6); (2) SAH group (n = 10, 4 mice were excluded for death); (3) SAH + WBV group (n = 8, 2 mice were excluded for death). TUNEL staining and immunofluorescence staining were performed to observe apoptotic cells and analyze the expression of GFAP and Iba-1. Furthermore, the Western blot assay was conducted to analyze the protein level of cleaved Caspase-3. ELISA assay was used to detect IL-10, IL-18, and IL-1β.

2.2.3 Experiment 3

To demonstrate the long-term neuroprotective effects of WBV, Morris Water Maze (MWM) test and rotarod test were performed to evaluate the memory and motor coordination ability of mice. 24 C57BL/6J male mice were randomly divided into 3 groups: (1) Sham group (n = 6); (2) SAH group (n = 13, 5 mice were excluded for death and 2 mice were excluded due to the SAH grade score lower than 8); (3) SAH + WBV group (n = 10, 2 mice were excluded for death and 2 mice were excluded due to the SAH grade score lower than 8). Rotarod test was carried out on the 7th, 14th, and 21st days after SAH induction. Additionally, the MWM test was performed on the 22nd to 27th day after SAH induction. On the 28th day after SAH induction, Nissl staining was conducted to evaluate the loss of neurons of the mice hippocampus.

2.3 SAH induction

As previously described [7], the endovascular perforation was performed to induce the mice's SAH model. After intubation and isoflurane (3%) anesthetization, skin dissection was performed from the cervical midline of mice. The muscle tissues and blood vessels were then isolated by blunt dissection. The left internal carotid artery was inserted with a 5-cm-long nylon suture (4–0, pointed) via the external carotid artery and carotid bifurcation. The suture was not discontinued until the resistance was felt at the bifurcation. Immediately following the arterial perforation, the suture was pulled out, and the final suturing of the incision was done. Model validation was carried out on the SAH grade scale involving the anatomical-morphological outcomes of the murine cerebral zone. The same procedure (apart from arterial puncturing) was implemented on the sham group mice as well.

2.4 Assessment of SAH grade

Mice were euthanized after 24 h of SAH induction. As per the previously elaborated standards, the SAH grade scoring of mice was evaluated by a researcher who was unaware of the assigned grouping [8]. We partitioned the basal cistern into size areas and scored them separately between 0 and 3. The exclusion criterion of this study was mice having a SAH score of < 8.

2.5 Evaluation of neurobehavioral test

The murine neurological function was assessed after 24 h of SAH induction using a beam balance experiment and a modified Garcia scale [9]. The indices for neurological function evaluated by the modified Garcia synthetic score were comprised of locomotor activity, forelimb motion, wire cage climbing, trunk touch, and a touch of vibrissae. Each of the above parts was scored between 0 and 3. Meanwhile, the mice were arranged onto the beam for the beam balance experiment, and their walking distance within 1 min was recorded. A score of 0 indicated no walk and fall; a score of 1 indicated no walk but staying on beam; a score of 2 indicated walk but fall; a score of 3 indicated walk shorter than 20 cm; a score of 4 indicated walk exceeding 20 cm. The beam balance experiment was conducted in triplicates, and the mean of the resulting values was taken for comparative analysis.

2.6 Evaluation of brain edema

Following a previously described protocol [10], the cerebral moisture content was measured to determine brain edema using the wet-dry technique after 24 h of SAH. After the mice were sacrificed, their left hemispheres were harvested and instantly weighed to record the wet weights. Next, the brain samples were subjected to 100°C for 24 h to obtain the dry weights. The computational formula used for measuring cerebral moisture content was as follows: (wet weight – dry weight)/wet weight × 100%.

2.7 TUNEL staining

We performed TUNEL staining to observe apoptosis as per the protocol of the manufacturer (Roche Inc., Switzerland). After isoflurane (3%) anesthetization and 24 h of SAH, transcardial perfusion was performed on the mice using ice-cold PBS (60 ml) and paraformaldehyde (60 ml, 10%) subsequently. Next, the brains were harvested and immobilized in paraformaldehyde (10%) for 24 h and maintained for an extra 72 h in sucrose solution (30%). After the incubation, brains were sectioned into 10 µm thick coronal pieces. The assessment site was the left temporal cortex present in the vicinity of the subarachnoid hematoma and hippocampus. 6 sections of each brain tissue were observed under fluorescence microscopy using high magnification. The counting for TUNEL-positive cells was accomplished by Image J software and was measured in terms of cells/mm². The data were represented as percentages (%) of neurons positive to TUNEL.

2.8 Immunofluorescence staining

4 µm thick brain slices were treated with 0.1% Triton X-100 and then blocked with 5% BSA. The samples were cultured with the following primary antibodies: Iba-1 (1:100, #17198, Cell Signaling), GFAP (1:100, #80788, Cell Signaling) overnight at 4 ° C. After 3 times of PBST washing, the samples were incubated with secondary antibodies at 37 ° C for 1 hour. The images were taken by light microscope (BX 53, Japan).

2.9 Nissl staining

4 µm thick brain slices were soaked in 0.1% cresol violet for 2 min, dehydrated in the flex tissue specimen system, removed in xylene, and covered with paramount. The left hippocampus of mice were photographed under the light microscope (BX 53, Japan). Image J software was used to analyze and calculate the number of Nissl- positive cells. The data were expressed as the number of neurons per field.

2.10 Western blot assay

Western blot was conducted as per the previously mentioned protocol [11]. After 24 h of SAH, mice were euthanized and perfused using pH 7.4 ice-cold PBS (200 ml). Their left hemispheres were harvested rapidly and preserved in a − 80°C refrigerator until further experimental use. The RIPA lysis buffer (#9806S) from Cell Signaling (USA) was used to homogenize the cerebral samples, which was followed by 20-min of centrifugation at 12,000 r/min at 4°C. The resulting supernatants were mixed using loading buffer (#7722S) from the same company and then subjected to 10-min of boiling before use. To perform the electrophoresis, 50 µL of each protein sample was placed into the SDS-polyacrylamide gel lane. Then, the protein isolates were shifted onto nitrocellulose membranes and subjected to 2-h blockage using 5% skimmed milk. This was followed by 8-h overnight incubation with primary antibodies at 4°C. The following primary antibodies were used for this experiment: anti-cleaved Caspase-3 (1:1000, #9661T, Cell Signaling, USA) and anti-β-actin (1:1000, #3700S, Cell Signaling, USA). Finally, the membranes were rinsed thrice and subsequently incubated with a secondary antibody conjugated to HRP for 2-h incubation at ambient temperature. An X-radiography was utilized to detect the protein bands, and Image J software was used to quantify their intensity.

2.11 ELISA assay

After 24 h of SAH induction, mice were euthanized and the brain tissue of the left hemisphere was used to detect inflammatory cytokines, such as IL-10, IL-18, and IL-1β according to the method of using the ELISA Kit described by the manufacturer (BMS 625, BNS630, and BMS629, Invitrogen).

2.12 Morris Water Maze test

To evaluate the long-term neurological deficits caused by SAH induction, the MWM test was performed as previously described [12]. The MWM test was performed on the 22ed day after SAH to eliminate the influence of movement disorder and/or sensory disorder on the test. The spatial acquisition trial was performed from 23rd to 26th day after SAH induction. On the 27th day after SAH induction, the mice were tested with a 60 s probe test and the platform was removed from the water.

2.13 Rotarod test

Rotarod test was performed to assess the motor and coordination abilities of mice as previously described [13]. The cylinder started at 5 and 10 revolutions per minute (rpm) and accelerated by 2 rpm every 5 s. The time that the mice could stay on the accelerating rotating cylinder was recorded by the beam circuit.

2.14 Statistical analysis

Experimental data were analyzed on GraphPad Prism 8.0 (La Jolla, USA) and were expressed as means ± SDs. Each group's data were examined for normality through Shapiro-Wilk and Kolmogorov-Smirnov tests. When data did not pass normality, the nonparametric test (Kruskal-Wallis test) was used for statistical analysis. To find the differences in the inter-group comparison, the one-way analysis of variance (ANOVA) was adopted, and subsequently, Tukey’s test of pairwise multiple comparisons was used. The P-values of 0.05 were considered significantly different.

3.1 SAH grade and animal usage

As shown in Table 1, 79 mice in total were involved in our study. 19 mice in the SAH group and SAH + WBV group were dead. 6 mice were excluded due to SAH grade scores lower than 8. The mortality rate of the mice in this study is 24.05% (19/79). There was no statistical difference in SAH grade scores between the SAH group and SAH + WBV group (p > 0.05, Fig. 1C).

3.2 WBV attenuated EBI after SAH

All the mice in this study were recieved WBV pretreatment, the body weight of these mice suggested that WBV can reduce brain water content (p < 0.05, Fig. 1D) without significantly affecting the weight of mice (p < 0.05, Fig. 1B). The modified Garcia scores and beam balance scores of mice in the SAH + WBV group were significantly increased than that in the SAH group (p < 0.05, Fig. 1E and F).

3.3 WBV alleviated neuronal apoptosis in the cerebral cortex of mice

WBV treatment also alleviated the SAH-induced neuronal apoptosis in the WBV treatment group than that in the SAH group (p < 0.05, Fig. 2A and B). The apoptosis-related protein (cleaved Caspase-3) level was also decreased in the WBV treatment group compared with the SAH group (p < 0.05, Fig. 2C and D).

3.4 WBV inhibited the expression of Iba-1 in the cerebral cortex of mice

The expression levels of Iba-1 were decreased in the WBV treatment group compared with the SAH group at 24 h after SAH (p < 0.05, Fig. 3A and B).

3.5 WBV inhibited the expression of GFAP in the cerebral cortex of mice

The immmunostaining also suggested that WBV treatment significantly decreased the expression of GFAP in the cerebral cortex of mice at 24 h after SAH (p < 0.05, Fig. 4A and B).

3.6 The effects of WBV on regulating inflammatory cytokines

ELISA assay was conducted to detect the levels of inflammatory cytokines, we found that the levels of IL-18, and IL-1β were downregulated by WBV. Additionally, the level of anti-inflammatory cytokine (IL-10) was increased in the WBV treatment group than that in the SAH group, which illustrated that WBV treatment has the effect of anti-inflammation at 24 h after SAH (p < 0.05, Fig. 5A, B and C).

3.7 WBV improved the motor ability of mice after SAH

Rotarod test was conducted in the 1st, 2ed and 3rd week after SAH induction to evaluate the motor ability of mice, our results revealed that WBV treatment improved the coordination, balance and motor ability of mice (p < 0.05, Fig. 6A).

3.8 WBV attenuated learning and memory deficits of mice after SAH

MWM test was conducted to evaluate the learning and memory ability of mice. The results demonstrated that the latency to platform decreased from day 1 to day 5, which reflects the learning process of mice. During the 5-day test, the time between the WBV treatment group and the SAH group was statistically different (p < 0.05, Fig. 6B). Furthermore, the time in platform quadrant was also longer in the WBV treatment group than that in the SAH group, indicating the improvement of learning and memory ability (p < 0.05, Fig. 6C).

3.9 WBV reduced the loss of neurons in the hippocampus of mice

Nissl staining was conducted to evaluate the neuronal loss of the hippocampus of mice. our results indicated that the number of Nissl-positive cells in the WBV treatment group was increased compared with the SAH group (p < 0.05, Fig. 6D and E), which illustrated that WBV treatment reduced the neuronal loss after SAH.

In the present study, we found that the SAH-induced neurological deficits of the WBV pre-treated mice, such as motor function, brain edema, and neuronal apoptosis, were significantly abated compared with the SAH group. WBV treatment decreased the expression levels of Iba-1 and GFAP. Additionally, inflammatory cytokines were regulated by WBV. The expression levels of IL-18 and IL-1β were downregulated, while the expression level of IL-10 was upregulated in the SAH + WBV group than that in the SAH group, which indicated that WBV may have the effects of alleviating neuroinflammation. The long-term neurological functions of mice were evaluated by the MWM test and rotarod test, which showed that the memory and learning deficits caused by SAH were attenuated and the coordination abilities of mice were improved by WBV. Lastly, Nissl staining was performed to observe the loss of neurons in the hippocampus of mice. Our results verified that the reduced number of Nissl-positive cells caused by SAH was abolished by WBV and this may be one of the mechanisms by which WBV improves the learning and memory function of mice.

At this stage, when the research of neuroprotective drugs encounters a bottleneck, physical exercise, as a relatively affordable intervention, plays a pretty significant role in CNS diseases. Relevant studies have shown that exercise intervention has a positive impact on memory function and motor function [14] [4]. WBV is an effective way of passive activity for some people who can not exercise actively due to physical or psychological diseases [15] [16]. In recent years, many studies have clarified the positive effects of WBV on various diseases. WBV has been proved to prevent bone loss caused by breast cancer (BC) metastasis and bone marrow vascular involvement. [17]. Soohrabzadeh et al. clarified that WBV can significantly reduce the plantar pain of diabetes patients and improve their motor coordination and their quality of life [18]. Furthermore, Pen et al. confirmed that WBV improved depression-like behavior in rats [19]. Here, the WBV at 30 Hz was used as an intervention based on some proven studies showing that WBV of 30 Hz has positive effects on the improvement of neurological function in mice as well as humans. Oroszi et al. proved that WBV at 30 Hz improves the cognitive abilities of mice [4]. Keijser et al. also found that WBV (30 Hz; 1.9G) significantly improves motor performance and attention for object recognition in mice [14]. In our study, the neuroprotective effects of WBV (30 Hz twice per day for 20 days) on neuroinflammation, neuronal apoptosis, and learning and memory deficit caused by SAH induction were verified. Therefore, rational use of this intervention may provide a new strategy for clinical treatment of SAH.

In previous studies, the roles of WBV in CNS diseases have been elucidated, but these studies mainly focus on the exploration of physical performance, cognitive ability, and mental state. Zhang et al. clarified that WBV treatment can improve the motor ability of stroke patients by improving nerve remodeling [20]. Wei & Cai verified that the physical performance and balance ability of patients with stroke were significantly improved by WBV [21]. Some studies have also shown that WBV can increase the expression level of neurotrophic factors and thus play a neuroprotective role. Raval et al. illustrated that the expression levels of brain-derived growth factor (BDNF) and TrkB protein were upregulated by WBV treatment [5]. Thus, it plays a neuroprotective role in ischemic cerebrovascular disease. Moreover, Ribeiro et al. confirmed that the blood BDNF levels of patients with fibromyalgia syndrome (FMS) were also increased by WBV treatment [22]. Similarly, Simao et al. found that the lower limb muscle performance of aged females with osteoarthritis of the knee (KOA) was improved by WBV treatment and the increased expression levels of BDNF may be involved in the underlying mechanism of the neuroprotective effects of WBV [23]. However, the therapeutic effects of WBV on neuroinflammation are rarely reported. Recently, Chen et al. confirmed that WBV treatment reduced TBI-induced brain damage by regulating neuroinflammation. Therefore, we proposed and confirmed the hypothesis that WBV can attenuate SAH-induced neuroinflammation and thus play a neuroprotective role in SAH.

SAH is one of the most dangerous cerebrovascular diseases worldwide, and has a high mortality and disability rate. The prognosis of patients with SAH has been proved to be closely related to neuroinflammation. In the acute stage of SAH, the inflammatory reaction is mainly manifested as the local inflammatory reaction caused by blood components entering the subarachnoid space and triggering the downstream inflammatory cascade. Subsequently, in the subacute and chronic phases, when central resident immune cells are activated, a large number of peripheral inflammatory cells enter the subarachnoid space under the chemotaxis of inflammatory cytokines [24]. Inflammatory cells secrete a variety of inflammatory cytokines and play an important pathogenic role in CNS diseases [25] [26]. Therefore, alleviating neuroinflammation may be one of the keys to attenuating the brain damage caused by SAH. In the present study, we found that WBV can decrease the expression levels of Iba-1 and GFAP compared with the SAH group. The expression levels of IL-1β and IL-18 were significantly decreased while the expression levels of IL-10 were increased in the SAH + WBV group than that in the SAH group. In addition to the EBI within 72 h after SAH, the impairment of learning and memory function is also one of the important factors affecting the prognosis of patients with SAH [27]. In this study, less neuronal loss of hippocampus in CA1, CA3, and DG regions was found in the SAH + WBV group compared with the SAH group, which suggested that WBV treatment contributes to the increased number of neurons in the hippocampus after SAH.

Some limitations to this study should be pointed out. Firstly, the WBV treatment in this study was achieved by pre-treatment, which indicated that the experimental mice in this study received WBV treatment before SAH-induction. However, clinical SAH is often acute and difficult to predict. Secondly, some detections of the neurotrophic factors that have been proven to be upregulated by WBV treatment were not involved in this study, which requires further exploration in the future. Lastly, the underlying mechanisms of the neuroprotecitve effects of WBV still need to be further investigated in the future.

To sum up, the neuroprotective effects of WBV were verified in our study. WBV at 30 Hz for 20 days reduced the EBI after SAH. The expression levels of Iba-1 and GFAP in the cerebral cortex and hippocampus of mice were markedly decreased in the SAH + WBV group than that in the SAH group, which suggested that WBV treatment attenuated the neuroinflammation caused by SAH. Furthermore, the long-term neurological dysfunctions of mice, such as learning and memory deficits, were significantly improved by WBV, which may be closely related to the less loss of neurons in the hippocampus of mice. These results suggested that WBV may be one of the potential candidates for the clinical treatment of SAH.

Author contributions

Yue Wang: experimental design, preparation of the first draft; Yingying Ding: animal experiments and molecular biology experiments; Wang Zhang: data analysis; Yu Sheng: image editing; Tao Chen: reviewing and proof reading; Yuhai Wang: reviewing and final proof reading.

Funding

This study was supported by the National Natural Science Foundation of China (No. 8187080447).

Conflict of interests

None

Zhang X, Karuna T, Yao ZQ, Duan CZ, Wang XM, Jiang ST, Li XF, Yin JH, He XY, Guo SQ, Chen YC, Liu WC, Li R, Fan HY (2018) High wall shear stress beyond a certain range in the parent artery could predict the risk of anterior communicating artery aneurysm rupture at follow-up. J Neurosurg 131:868–875
Sehba FA, Hou J, Pluta RM, Zhang JH (2012) The importance of early brain injury after subarachnoid hemorrhage. Prog Neurobiol 97:14–37
Huang D, Yang Z, Wang Z, Wang P, Qu Y (2018) The macroscopic and microscopic effect of low-frequency whole-body vibration after cerebral ischemia in rats. Metab Brain Dis 33:15–25
Oroszi T, Geerts E, de Boer SF, Schoemaker RG, van der Zee EA, Nyakas C (2021) Whole Body Vibration Improves Spatial Memory, Anxiety-Like Behavior, and Motor Performance in Aged Male and Female Rats. Front Aging Neurosci 13:801828
Raval AP, Schatz M, Bhattacharya P, d'Adesky N, Rundek T, Dietrich WD, Bramlett HM (2018) Whole Body Vibration Therapy after Ischemia Reduces Brain Damage in Reproductively Senescent Female Rats.Int J Mol Sci19
Chen T, Liu WB, Ren X, Li YF, Li W, Hang CH, Wang YH (2022) Whole Body Vibration Attenuates Brain Damage and Neuroinflammation Following Experimental Traumatic Brain Injury. Front Cell Dev Biol 10:847859
Xu P, Tao C, Zhu Y, Wang G, Kong L, Li W, Li R, Li J, Zhang C, Wang L, Liu X, Sun W, Hu W (2021) TAK1 mediates neuronal pyroptosis in early brain injury after subarachnoid hemorrhage. J Neuroinflammation 18:188
Sugawara T, Ayer R, Jadhav V, Zhang JH (2008) A new grading system evaluating bleeding scale in filament perforation subarachnoid hemorrhage rat model. J Neurosci Methods 167:327–334
Fan R, Enkhjargal B, Camara R, Yan F, Gong L, ShengtaoYao, Tang J, Chen Y, Zhang JH (2017) Critical role of EphA4 in early brain injury after subarachnoid hemorrhage in rat. Exp Neurol 296:41–48
Okada T, Enkhjargal B, Travis ZD, Ocak U, Tang J, Suzuki H, Zhang JH (2019) FGF-2 Attenuates Neuronal Apoptosis via FGFR3/PI3k/Akt Signaling Pathway After Subarachnoid Hemorrhage. Mol Neurobiol 56:8203–8219
Li Y, Wang X, Liu M, Zhang W, Li R, Nie Z (2020) Effect of Methylene Blue and PI3K-Akt Pathway Inhibitors on the Neurovascular System after Chronic Cerebral Hypoperfusion in Rats. J Mol Neurosci
Bromley-Brits K, Deng Y, Song W (2011) Morris water maze test for learning and memory deficits in Alzheimer's disease model mice. J Vis Exp
Xie Z, Enkhjargal B, Wu L, Zhou K, Sun C, Hu X, Gospodarev V, Tang J, You C, Zhang JH (2018) Exendin-4 attenuates neuronal death via GLP-1R/PI3K/Akt pathway in early brain injury after subarachnoid hemorrhage in rats. Neuropharmacology 128:142–151
Keijser JN, van Heuvelen MJG, Nyakas C, Toth K, Schoemaker RG, Zeinstra E, van der Zee EA (2017) Whole Body Vibration Improves Attention and Motor Performance in Mice Depending on the Duration of the Whole-Body Vibration Session. Afr J Tradit Complement Altern Med 14:128–134
van Heuvelen MJG, Rittweger J, Judex S, Sanudo B, Seixas A, Fuermaier ABM, Tucha O, Nyakas C, Marin PJ, Taiar R, Stark C, Schoenau E, Sa-Caputo DC, Bernardo-Filho M, van der Zee EA (2021) Reporting Guidelines for Whole-Body Vibration Studies in Humans, Animals and Cell Cultures: A Consensus Statement from an International Group of Experts. Biology (Basel) 10
Bemben D, Stark C, Taiar R, Bernardo-Filho M (2018) Relevance of Whole-Body Vibration Exercises on Muscle Strength/Power and Bone of Elderly Individuals. Dose Response 16:1559325818813066
Matsumoto T, Mukohara A (2022) Effects of Whole-Body Vibration on Breast Cancer Bone Metastasis and Vascularization in Mice. Calcif Tissue Int
Sohrabzadeh E, Kalantari KK, Naimi SS, Daryabor A, Akbari NJ (2022) The immediate effect of a single whole-body vibration session on balance, skin sensation, and pain in patients with type 2 diabetic neuropathy. J Diabetes Metab Disord 21:43–49
Peng G, Yang L, Wu CY, Zhang LL, Wu CY, Li F, Shi HW, Hou J, Zhang LM, Ma X, Xiong J, Pan H, Zhang GQ (2021) Whole body vibration training improves depression-like behaviors in a rat chronic restraint stress model. Neurochem Int 142:104926
Zhang M, Wei J, Wu X (2022) Effects of whole-body vibration training on lower limb motor function and neural plasticity in patients with stroke: protocol for a randomised controlled clinical trial. BMJ Open 12:e060796
Wei N, Cai M (2022) Optimal frequency of whole body vibration training for improving balance and physical performance in the older people with chronic stroke: A randomized controlled trial. Clin Rehabil 36:342–349
Ribeiro VGC, Lacerda ACR, Santos JM, Coelho-Oliveira AC, Fonseca SF, Prates ACN, Flor J, Garcia BCC, Tossige-Gomes R, Leite HR, Fernandes JSC, Arrieiro AN, Sartorio A, Sanudo B, Sa-Caputo DC, Bernardo-Filho M, Figueiredo PHS, Costa HS, Lima VP, Cardoso RF, Bastone AC, Soares LA, Mendonca VA, Taiar R (2021) Efficacy of Whole-Body Vibration Training on Brain-Derived Neurotrophic Factor, Clinical and Functional Outcomes, and Quality of Life in Women with Fibromyalgia Syndrome: A Randomized Controlled Trial. J Healthc Eng 2021:7593802
Simao AP, Mendonca VA, Avelar NCP, da Fonseca SF, Santos JM, de Oliveira ACC, Tossige-Gomes R, Ribeiro VGC, Neves CDC, Balthazar CH, Leite HR, Figueiredo PHS, Bernardo-Filho M, Lacerda ACR (2019) Whole Body Vibration Training on Muscle Strength and Brain-Derived Neurotrophic Factor Levels in Elderly Woman With Knee Osteoarthritis: A Randomized Clinical Trial Study. Front Physiol 10:756
Zhou Y, Wang Y, Wang J, Anne Stetler R, Yang QW (2014) Inflammation in intracerebral hemorrhage: from mechanisms to clinical translation. Prog Neurobiol 115:25–44
Hu X, Tao C, Gan Q, Zheng J, Li H, You C (2016) Oxidative Stress in Intracerebral Hemorrhage: Sources, Mechanisms, and Therapeutic Targets. Oxid Med Cell Longev 2016:3215391
Healy LM, Yaqubi M, Ludwin S, Antel JP (2020) Species differences in immune-mediated CNS tissue injury and repair: A (neuro)inflammatory topic. Glia 68:811–829
Regnier-Golanov AS, Gulinello M, Hernandez MS, Golanov EV, Britz GW (2022) Subarachnoid Hemorrhage Induces Sub-acute and Early Chronic Impairment in Learning and Memory in Mice. Transl Stroke Res 13:625–640

Tables 1 is available in the Supplementary Files section.

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Whole-body vibration protects against early brain injury and neuroinflammation after experimental subarachnoid hemorrhage

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Abstract

Figures

1. Introduction

2. Materials And Methods

2.1 Experimental animals

2.2 Experimental design

2.2.1 Experiment 1

2.2.2 Experiment 2

2.2.3 Experiment 3

2.3 SAH induction

2.4 Assessment of SAH grade

2.5 Evaluation of neurobehavioral test

2.6 Evaluation of brain edema

2.7 TUNEL staining

2.8 Immunofluorescence staining

2.9 Nissl staining

2.10 Western blot assay

3. Results

3.1 SAH grade and animal usage

3.2 WBV attenuated EBI after SAH

3.3 WBV alleviated neuronal apoptosis in the cerebral cortex of mice

3.4 WBV inhibited the expression of Iba-1 in the cerebral cortex of mice

3.5 WBV inhibited the expression of GFAP in the cerebral cortex of mice

3.6 The effects of WBV on regulating inflammatory cytokines

3.7 WBV improved the motor ability of mice after SAH

3.8 WBV attenuated learning and memory deficits of mice after SAH

3.9 WBV reduced the loss of neurons in the hippocampus of mice

4. Discussion

5. Conclusion

Declarations

References

Tables

Additional Declarations

Supplementary Files

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