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BACKGROUND: Human umbilical cord mesenchymal stem cells play a vital role in the repair of the blood-brain barrier after traumatic brain injury. OBJECTIVE: To investigate the protective effect of human umbilical cord blood mesenchymal stem cell transplantation on the blood-brain barrier after traumatic brain injury in rats and its possible mechanism. METHODS: Sixty Sprague-Dawley rats were randomly divided into sham operation group, injury control group (model group), cell transplantation group and Sunitinib group, with 15 rats in each group. Traumatic brain injury model was established by improved hydraulic impact method in all the groups except for the sham operation group. Rats in the sham operation group and model group were injected with 1 mL of normal saline, and those in the cell transplantation group were injected with 1 mL of 2×109/L human umbilical cord blood mesenchymal stem cells. The injection was done via the tail vein at 0.5, 24, and 48 hours after modeling. In the Sunitinib inhibitor group, the rats were given oral PDGFR-β pathway inhibitor, Sunitinib (80 mg/kg), from 1 day before modeling until being executed. Three days after modeling, the water content in brain tissue was measured by dry-wet specific gravity method, the permeability of the blood-brain barrier was measured by Evans blue method, expression of GFAP and vWF was observed by immunofluorescence staining and the expression of blood-brain barrier related proteins and PDGFR-β pathway proteins was detected by western blot method. RESULTS AND CONCLUSION: Compared with the sham operation group, the brain water content of the model group increased significantly (P < 0.05), while that of the cell transplantation group was significantly lower than that of the model group (P < 0.05). The Evans blue content in the model group was significantly higher than that in the sham operation group (P < 0.05), while the Evans blue content in the cell transplantation group was significantly lower than that in the model group (P < 0.05). Compared with the sham operation group, the expression of vWF and GFAP increased significantly in the model group (P < 0.05), while compared with the model group, the expression was significantly reduced in the cell transplantation group (P < 0.05). Western blot showed that ZO-1, Oclaudin-5, and PDGFR-β protein expressions in the model group were significantly lower than those in the sham operation group (P < 0.05), while these expressions were significantly increased in the cell transplantation group as compared with the model group (P < 0.05). To conclude, intravenous injection of human umbilical cord mesenchymal stem cells through the tail ein can reduce the permeability of blood-brain barrier and play a neuroprotective role in rats with traumatic brain injury. Its possible mechanism is related to the promotion of PDGFR-β expression in the injured area.
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Objective The nerve-protective effect of Apoli-poprotein J ( ApoJ) in intracerebral hemorrhage ( ICH) is not yet clarified.This study aimed to explore the therapeutic effect of trans -plantation of bone marrow mesenchymal stem cells (BMSCs) carrying the ApoJ gene on intracerebral hemorrhage in rats and its possible ac -tion mechanism. Methods Rat BMSCs were isolated, cultured in vitro, and transfected with the recombinant plasmid pEGFP -N1-ApoJ mediated with lipofectamine.Ninety-six SD rats were randomly divided into groups A, B and C, and ICH models were established by two -step autologous intracranial blood injection .At 24 hours after model-ing, the rats in groups A, B, and C were transplanted with the same volume of ApoJ-transfected BMSC suspension, BMSC suspension and normal saline, respectively.At 1, 3, 5 and 7 days after transplantation, the neurofunction recovery of the rats were evaluated with modified Neurological Severity Scores (mNSS), the brain water content measured by the dry -wet weight method, and the expression level of complement component 3 (C3) in the brain tissue detected by Western blot . Results The mNSS exhibited no statistically significant differences among the three group of rats on the 1st day after transplantation (P>0.05), but was remarkably lower in group A than in B and C on the 3rd (8.13±0.99 vs 9.25±1.28 and 10.88±0.84, P<0.05), 5th (6.75±1.04 vs 8.50±1.41 and 9.75±0.89, P<0.05) and 7th day (5.63±0.52 vs 7.00±0.54 and 7.88±1.25, P<0.05), and markedly lower in group B than in C (P<0.05).The water content in the brain tissue was also significantly lower in group A than in B and C on the 1st (78.17±0.82 vs 78.83±0.56 and 80.38±0.35, P<0.05), 3rd (78.68±0.55 vs 79.12±0.26 and 81.47±0.26, P<0.05), 5th (77.00±0.58 vs 78.13±0.46 and 79.74± 0.41, P<0.05) and 7th day (75.89±0.46 vs 76.86±0.29 and 78.44±0.44, P<0.05), and remarkably lower in group B than in C (P<0.05).Western blot showed that the expression of the C 3 protein in the brain tissue was markedly decreased in group A as compared with B and C on the 1st (0.096±0.011 vs 0.212±0.014 and 0.440±0.006, P<0.05), 3rd (0.083±0.005 vs 0.164±0.013 and 0.604± 0.011, P<0.05), 5th (0.064±0.009 vs 0.105±0.010 and 0.333±0.010, P<0.05), 7th day (0.045±0.007 vs 0.091±0.004 and 0.141± 0.003, P<0.05), and significantly lower in group B than in C (P<0.05). Conclusion ApoJ can promote the recovery of the neuro-logical function of ICH rats by inhibiting complement activation -mediated secondary brain damage and alleviating cerebral edema .
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Objective To investigate the effects of minimally invasive intracranial hematoma clearance on the perihematomal glutamate(Glu) level,permeability of blood-brain barrier(BBB) and brain edema.Methods Thirty rabbits with body weight of 2.80-3.40 kg were used to established the model of spontaneous intracerebral hemorrhage(ICH) and randomly divided into the minimally invasive group(MI) and control group(MC) after the model was prepared successfully.The MI group underwent minimally invasive procedures for removing intracranial hematoma by stereotactic instrument within 6 h after establishing the ICH model.The brain tissue was extracted on postoperative 1,3,7 d,and the perihematomal brain tissues were taken to detect the Glu level,BBB permeability and water content of brain tissue,which were compared with those in the control group.Results The Glu level,BBB permeability and brain water content on 1,3,7 d in the MI group were lower than those in the MC group,and the differences were statistically significant(P＜0.05).Conclusion The minimally invasive surgery for removing intracranial hematoma is helpful to reduce perihematoma Glu level,BBB permeability and brain water content.
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OBJECTIVE: To study the protective effects of sparstolonin B(8,5'-dihydroxy-4-phenyl-5,2'-oxidoisocoumarin, SsnB) on mouse with intracerebral haemorrhage (ICH). METHODS: The effect of SsnB on neuronal cell death induced by Hb using a microglia-neuron transwell system was investigated. Autologous nonanticoagulated blood was collected from the tail artery of the mouse and then injected into the striatum using a syringe pump. Neurological deficit of the mouse with ICH was assessed 1,3,5 and 7 d after SsnB administration using modified neurological severity score system, and brain water content of the mouse cerebral tissues after ICH was measured at the same day. The concentrations of pro-inflammatory cytokines in perihematoma tissues, including interleukin-6, interleukin-1 p and tumor necrosis factor-α, were measured by ELISA assay. RESULTS: The results show that SsnB significantly reduced the neurological deficit scores and the brain water content, and inhibited the levels of IL-6, IL-1β and TNF-α at first day and third day after ICH. CONCLUSION: SsnB can improve the brain injury in mouse induced by ICH. The mechanism may involve increased the neuronal survival rate and decreased expression of pro-inflammatory cytokines of mouse after ICH.
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Objective To explore the protective effect of 17β-estradiol on traumatic brain injury in rats.Methods A total of 45 adult male SD rats were divided into 3 groups using the random digit table, 15 rats in each group:the control group only exposed but not injured the brain, the injury group received traumatic brain injury ( TBI) by Feeney’ s method, and the treatment group received the same handling with injury group, and pretreatment with 17β-estradiol peritoneal injection, 1 mg/kg per day for one week.The other two groups were given the same volume of castor oil.At 6 h, 24 h and 48 h after injury, the brain tissue water content, malondialdehyde ( MDA) content and superoxide dismutase ( SOD) activity were measured.Results At 6 h, 24 h and 48 h after injury, the levels of brain tissue water content in the injury group and treatment group were significantly higher than those in the control group ( P0.05).At 24 h and 48 h after injury, the brain tissue water contents in the injury group was ( 105.17 ±0.43 )% and ( 107.54 ±0.39 )%, in the treatment group was (103.26 ±0.42)%and (105.89 ±0.43)%, respectively, showing a significant difference between the two groups ( P0.05).But at 24 h and 48 h after injury, the levels of MDA in the treatment group were significantly lower than those in the injury group [(130.39 ±7.02) μmol/g vs.(149.41 ±8.25) μmol/g, (125.41 ±6.59) μmol/g vs.(157.72 ± 8.93) μmol/L] , and the levels of SOD in the treatment group were significantly higher than thoset in the injury group [(88.46 ±7.17) U/g vs.(80.10 ±4.87) U/mg, (97.31 ±7.89) U/g vs.(84.29 ±6.13) U/g], with a significant difference ( P<0.05 ) between the two groups.Conclusions 17β-estradiol has a protective effect on traumatic brain injury.
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OBJECTIVE: To study the effects of anti-inflammatory active fraction (AF) from the tuber of Scirpus yagaraohwi in mice with intracerebral haemorrhage (ICH). METHODS: A 25 μL volume of autologous nonanticoagulated blood was collected from the tail artery of the mouse and then injected into the striatum using a syringe pump. Neurological deficits of the mouse was assessed 1, 3 and 5 days after ICH using a 28-point neurological deficit scale, and brain water content of the mouse cerebral tissues after ICH was measured at third day.The concentrations of pro-inflammatory cytokines in perihematoma tissues, including interleukin-6, interleukin-1β and tumor necrosis factor-α, were measured by ELISA assay. RESULTS: The results show that AF significantly reduced the neurological deficit scores and the brain water content after ICH. AF also markedly inhibited the levels of IL-6, IL-1β and TNF-α at first day and third day alter ICH. CONCLUSION: Anti-inflammatory active fraction of Scirpus yagaraohwi can improve the brain injury in mouse induced by ICH. The mechanism may involve decreased expression of pro-inflammatory cytokines of mouse after ICH.
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Objective To study the establishment of rat model of asphyxia-cardiac arrest and efficacy of CPR in order to find the length of optimum time of asphyxia to cause injury.Methods One hundred and twenty-six male Sprague-Dawley rats were randomly (random number) divided into sham operation group and experimental groups.Cardiac arrest was induced by asphyxiation after intravenous injection of vecuronium bromide.The experimental groups were assigned into AP4 (four-minute asphyxia period),AP6 and AP8 subgroups in accordance with different lengths of time of asphyxia subjected to.In these groups,CPR,including pre-cordial compression and synchronized mechanical ventilation,was initiated 4,6 and 8 min after asphyxia-induced cardiac arrest,respectively.The successful ratio of resuscitation and hemodynamic variables were recorded.Brain water content,neural deficit scores (NDS),imaging changes on MR,pathological changes of brain tissue and neuronal apoptosis were evaluated at 1 d,3 d and 7 days after ROSC.All the data were analyzed by single-factor analysis of variance or Chi-square test.P ＜ 0.05 was considered statistically significant.Result The lowest NDS occurred at 1 d after ROSC,brain water content and imaging changes on MR were most obvious at 3 d after ROSC,while pathological changes of brain tissue and neuronal apoptosis increased and reached the peak at 7d after ROSC.The survival rates after 24 hours of AP4,AP6 and AP8 groups were 85％,75％ and 45％,respectively.The rate of ROSC and survival rate of AP8 group were significantly lower than those of other groups (P ＜0.01).The longer time of asphyxia the severer pathological changes of brain tissue,brain edema,neural deficit,and magnetic resonance imaging changes in all experimental groups.As compared to other groups,the brain damage index of AP8 group was most serious,while that of AP6 group was moderate.Conclusions The rat model following asphyxia-induced cardiac arrest and cardiopulmonary resuscitation was established successfully.From the evidence of survival rate and damage grade of brain tissue,asphyxia for 6 min may be the rational length of ischemic time in this model.
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This study was aimed to explore the effect of Catalpol on the cerebral infarction size in acute phase, water content and inflammatory reaction of early recovery after permanent middle cerebral artery occlusion (pM-CAO). Adult male Sprague-dawley rats were subjected to chemical method to establish MCAT model. The detec-tion was made on neurobehavioral symptoms, cerebral infarction volume and water content at 24 h after surgery. The content of intedeukin-6 (IL-6), intedeukin-10 (IL-10) and nuclear factor kappa Bp65 (NF-κBp65) were de-tected after pMCAO with enzyme-linked immunosorbent assay (ELISA). The results showed that 24 h after MCAT, Catalpol 15-60 mg·kg-1 can significantly improve the neurobehavioral symptoms (P < 0.01, or P <0.001). The Catalpol 15 mg·kg-1 can significantly reduce cerebral infarction volume (P < 0.05). The Catalpol 30-60 mg·kg-1 can significantly reduce water content (P < 0.05). Catalpol 30-60 mg·kg-1 can significantly improve neurobehav-ioral symptoms from the 7th day after pMCAO. On the 14th after pMCAO, the content of IL-10 and NF-κBp65 of ischemia brain had no difference compared with sham-operated group. The IL-6 level of ischemia brain was obvi-ously reduced than the sham-operated group(P < 0.05). The intragastric administration of Catalpol 15 mg·kg-1 for 14 days can reduce the content of NF-κBp65 in the ischemia brain of model rats (P < 0.05). It was concluded that Catalpol can improve neurobehavioral symptoms of acute and subacute phase after cerebral ischemia, reduce infarcts and water content. These effects may not be related with its inhibition of inflammatory derived from cere-bral ischemia.
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Objective To observe the effect of splenectomy on mortality and brain water content of rats with brain injury so as to explore novel way for better clinical management of patients with severe traumatic brain injury. Methods Adult male SD rats were randomly divided into three groups, ie, sham operation on brain and spleen (Group A, n = 23), experimental brain trauma & sham operation on spleen (Group B, n =48) and experimental brain injury & splenectomy (Group C, n = 47). Modified Feeney' s method was used to create the animal model of experimental brain trauma, Longa' s scale was applied to evaluate the neurologic defect. Mortality within seven days following brain injury was calculat-ed. In the meantime, the brain water content was detected at days 1 (n = 8), 2 (n = 8), 3 (n = 8) and 7 (n = 7) after brain injury in each group, Results No statistical difference of Longs' s scale was found between Group B and Group C (P > 0.05). The mortalities within seven days after brain injury were 0%, 35.42 and 14.89% in Groups A, B and C respectively, with statistical difference between groups (P<0.05). The brain water content of Groups B and C at days 1, 2, 3 and 7 were (81.98±0.35)% & (81.78±0.41)%, (82.58±0.63)% & (81.81±0.48)% (P<0.05),(82.54±0.54)% & (81.52±0.84)% (P<0.05) and (81.50±0.41)% & (81.21±0.36)% (P>0.05) respectively. Conclusion Splenectomy can effectively reduce brain water content and significantly decrease mortality in rata with brain injury.
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Objective To observe the neuroprotection of Baicalin on brain damage following experimental intracerebral hemorrhage in rats.Methods Rat model of intracerebral hemorrhage was induced by collagenase method.The rats were randomly divided into sham operation control group,model group,high-,middle-and low-dose drug group,treated with different dose of Baicalin.The brain water content and the level of SOD,MDA,NO and NOS were measured at the third and seventh day after modeling repectively.Results Compared with the sham operation control group,the brain water content and the level of MDA,NO and NOS were significantly higher,and the level of SOD was significantly lower in model group.The brain water content and the level of MDA,NO and NOS were significantly lower,and the level of SOD was obviously higher in high-,middle-and low-dose Baicalin group than that in model group.Conclusion Baicalin has evident protective effect on brain damage after experimental intracerebral hemorrhage in rats,which maybe related to the increase of SOD and decrease of MDA,NO and NOS.
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Objective: To investigate the effective of Sanhua decotion(三化汤) on early brain water content,penetration of blood brain-barrier and the expression of aquaporin4(AQP4) in rats with acute cerebral ischemia/reperfusion(I/R).Methods: The string inserting method was employed to reproduce the rat model of focal cerebral I/R SD rats were randomly divided into sham operation group,model group and Sanhua decotion group.After I/R,each group was then divided into 6 hours,12 hours,24 hours,48 hours and 72 hours five subgroups.The brain water content was detected by the ratio of wet/dry weight to evaluate cerebral edema and the blood-brain barrier(BBB) damage was observed by Evan′s blue(EB) staining.The AQP4 mRNA and protein expressions were measured by reverse transcription-polymerase chain reaction(RTPCR) and immunohistochemistry respectively.Results: Compared with sham operation group,the brain water content,EB content,AQP4 protein and AQP4 mRNA expressions were increased at every time point after I/R in the model group(P
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Objective To observe the protective effect of edaraven on brain tissue and the therapeutic mechanism in rats following intracerebral hemorrhage(ICH).Methods In this series,96 Sprague-Dawley rats were divided into three groups at random(n=32 in each group):sham-operative group,ICH model group and edaraven therapy group.ICH model was induced by infusion of autologous whole blood into the caudate-putamen.Rats in sham-operative group were treated the same as above,but no blood injected.Rats in edaraven therapy group were treated with edaraven(3 mg/kg)by peritoneal injection 30 min before operation and once per 12 hours after operation.These rats were killed at 6,24,72,and 168 h after neurologic impairment test with Longa criteria.Brain water content was measured by dry-wet weight method.Malondialdehyde and TNF-alpha in the vicinity of the hematoma were measured by thio-barbituric acid method and ELISA respectively.Results Compared with sham operative group,neurologic impairment score,brain water content,malondialdehyde and TNF-alpha in brain tissues were obviously higher in ICH model group(P
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The effects of propylene glycol mannate sulfate (PGMS) on the lipid peroxide (MDA) , superoxide dismutase (SOD) , glutathion peroxidase (GSH-Px), nitric oxide (NO)and water content in the whole cerebral ischemia/reperfusion injury rabbits induced by four-vessel occlusion,The results show that PGMS can decrease the brain water and MDA, and can increase the SOD and GSH-Px level. No significant effect on the NO level has been detected. The results suggest that the protective effects of PGMS on ischemia/reperfusion injury may be related to its antioxidation.
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Objective To study the effects of phycocyanin on the expressions of matrix metalloproteinase 2 and 9(MMP-2 and MMP-9) after focal cerebral ischemic reperfusion in rats.Methods A rat middle cerebral artery occlusion/reperfusion(MCAO/R) model was built up using the intraluminal filament method and treated by phycocyanin.The brain water content(BWC) was calculated with dry-wet method and the Evans blue(EB) of the brain was measured with the method of colorimetry.The expressions of MMP-2 and MMP-9 were determined by immunohistochemical assay.Results The BWC and EB of the brain increased after cerebral ischemic reperfusion and peaked at reperfusion 2d,and decreased after applying phycocyanin.In control group,the upregulation of MMP-2 began at ischemic reperfusion 24h,reached a maximum at 3～7d,then subsided gradually,but was still in high level at 14d.In phycocyanin group,the expressing time-phase pattern of MMP-2 was similar to and significantly lower than that in control group at same time.In control group,the upregulation of MMP-9 began at ischemic reperfusion 6h,increased at 12h and reached a maximum at 2d,then subsided gradually at 3d to sham group level at 14d.In the phycocyanin group,the expression of MMP-9 was significantly lower than that in control group,and the time-phase pattern of MMP-9 was similar to that in control group.Conclusions Phycocyanin might play an neuroprotective effect by down regulating the expressions of MMP-2 and MMP-9 to reduce brain edema after cerebral ischemic reperfusion.
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Objective To investigate the changes and correlation between nuclear factor-?B (NF-?B) expression and brain water content (BWC) in perihematoma after experimental intracerebral hemorrhage (ICH) in rats. Methods Experimental ICH models of rat were made by injecting autologous blood using stereotaxic method. The expression of NF-?B in cerebral tissues was detected by immunohistochemistry technique. At the due time, BWC was measurement by day-wet method.Results NF-?B expression increased obviously at 6 h and peaked at 48 h in ICH group(P
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Objective:To investigate the relationship between the dynamic plasma level of arginine-vasopressin (AVP) and the brain water content (BWC) in the rats after severe burn.Methods:Radioimmunoassay(RIA) was performed to measure the dynamic plasma levels of AVP in the rats.Dry-wet weight method was used to measure the changes of water content in brain.All the data were statistically processed.Results:The plasma level of AVP and BWC in the rats increased after severe burn,and the plasma level reached the peak at 6h after burn.The positive correlations were observed between the two parameters( r =0.8790, P