Changes of Ski expression levels in rat activated astrocytes

AIM: To explore the time-dependent change of Ski protein expression in normal and activated astrocytes in rats.METHODS: The astrocytes were obtained from rat cerebral cortex and cultured in vitro. The astrocytes were treated with LPS and scratch injury for activation. Western blot analysis was used to determine glial fibrillary acidic protein (GFAP) and Ski protein levels in activated astrocytes at a series of time points. The indirect immunofluorescence staining method was performed to detect the location of Ski protein in the astrocytes.RESULTS: The protein of GFAP was naturally expressed in the astrocytes, beginning to increase after treated with LPS and scratch injury. Little protein expression of Ski in the normal astrocytes was observed. The Ski protein expression began to increase after treated with 1 mg/L LPS, peaked at 4 d (P<0.05) and then deceased, but was stills higher than that in the normal cells. The protein expression level of Ski after scratch injury was highly consistent with above mentioned. Ski was mainly observed in the nucleus of the normal cells and the cells treated with LPS for 6 d, while it was observed in the cytoplasm 2 and 4 d after treated with LPS.CONCLUSION: The protein of Ski is expressed in the astrocytes, and the expression level is increased in activated astrocytes, mainly located in the nucelus. Ski may plays an essential roles in the processes of activation and proliferation of astrocytes.     

Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.     

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

Effects of betaine on expression of GFAP,glycine and glycine receptor in hippocampus of pentylenetetrazole-induced epileptic rats

AIM：To study the effects of betaine on glial fibrillary acidic protein (GFAP), glycine (Gly) and glycine receptor (GlyR) expression in the hippocampus of rats with epilepsy induced by pentylenetetrazole (PTZ). METHODS：Forty-eight healthy male Wistar rats were randomly divided into control group, PTZ (35 mg·kg-1·d-1, intraperitoneal injection) group, PTZ+betaine (450 mg·kg-1·d-1, intragastric administration) group, PTZ+betaine (225 mg·kg-1·d-1, intragastric administration) group, PTZ+betaine (112.5 mg·kg-1·d-1, intragastric administration) group and PTZ+sodium valproate (200 mg·kg-1·d-1, intragastric administration) group. The rats in control group were intraperitoneally injected with saline at the same volume as PTZ injection, and those in control group and PTZ group received intragastric administration of saline at 1.0 mL·d-1. Rat behavior was recorded. Serum homocysteine (Hcy) level was measured. The expression of GFAP in the hippocampus was measured by immunofluorescence. Hippocampal Gly content was measured by an amino acid analysis system. The expression of GlyR was detected by immunofluorescence and Western blotting. RESULTS：There was no difference in the latency of grand mal seizures among groups (P>0.05). However, betaine treatment significantly decreased the duration of the first grand mal seizure compared with PTZ group (P<0.01). Serum Hcy level in PTZ group was significantly lowered compared with control group (P<0.01), and further decreased after betaine treatment (P<0.05). GFAP in PTZ group was significantly higher than that in control group (P<0.01), and decreased after betaine treatment (P<0.05). Gly in PTZ group was significantly lowered compared with control group (P<0.01), and increased after betaine treatment (P<0.05). The content of GlyR among groups showed the same trend as Gly. CONCLUSION：Betaine treatment shows antiepileptic effect, which may be related to its effects on the metabolites of Hcy and Gly.     

Expression of glial fibrillary acidic protein in developing rat brain after intrauterine infection

AIM: To investigate the alteration of glial fibrillary acidic protein (GFAP) immunoreactivity in developing rat brain after intrauterine infection. METHODS: Escherichia coli (E.coli) was inoculated into uterine horn of pregnant rats when gestation was 15 days and the control group was inoculated with normal saline. Immunohistochemistry was used for evaluation of GFAP expression in pup brains at postnatal day 1 (P1), P3, P7, P14, P21. RESULTS: GFAP-immunopositive cells was scarce in the periventricular white matter at P1 and P3 in two groups (P>0.05), but not in other brain regions. The number of GFAP-immunopositive cells of the E.coli-treated pups was markedly increased in periventricular white matter and hippocampus at P7 compared with the control group (P<0.05). The E.coli-treated pups at P14 showed a marked increase in GFAP expression in periventricular white matter, corpus callosum and cortex (P<0.01). However, no significant different levels of GFAP expression in any brain regions were found at P21 between two groups (P>0.05). CONCLUSION: Intrauterine infection induces an increased expression of GFAP in the neonatal brain.     

Losartan inhibits LPS-induced GFAP expression via AMPK activation in mouse hippocampus

AIM: To investigate the effects of losartan on lipopolysaccharide (LPS)-induced glial fibrillary acidic protein (GFAP) expression, and to determine whether adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activation is involved in the mechanism.METHODS: Adult male KM mice were divided into control group, LPS model group, losartan treatment group, and losartan and Compound C co-treatment group. To establish a model of central nervous system inflammation, the mice received daily intracerebroventricular injection of LPS (24 μg/d) for 2 d. Daily losartan administration (0.5, 1 or 5 mg·kg^-1·d^-1, ip) initiated at 14 d prior to LPS injection. Compound C (10 mg/kg, ip), a selective AMPK inhibitor, started to be injected daily at 2 d prior to LPS injection. The hippocampal tissues in each group were isolated at 3 d after the last LPS injection, and then the protein levels of GFAP, AMPK, p-AMPK, mammalian target of rapamycin (mTOR) and p-mTOR were determined by Western blot.RESULTS: Twice LPS injections significantly increased the expression of GFAP in the hippocampus (P<0.01). Losartan inhibited LPS-induced GFAP expression in a concentration-dependent way, and losartan at 5 mg·kg^-1·d^-1 significantly inhibited GFAP expression and AMPK activation (P<0.05), but it had no obvious effect on mTOR activation. Furthermore, Compound C significantly reversed the effect of losartan treatment on LPS-induced GFAP expression and AMPK phosphorylation (P<0.05).CONCLUSION: Losartan inhibits LPS-induced GFAP expression in the mouse hippocampus, and AMPK activation but not mTOR, is involved in the mechanism.     

Effects of astragaloside IV on autophagy in rats with cerebral ischemia/reperfusion injury

AIM: To investigate the effects of astragaloside IV (AS-IV) on autophagy in rats with cerebral ischemia/reperfusion (I/R) injury. METHODS: The focal cerebral ischemia/reperfusion of rat left middle cerebral artery occlusion (MCAO) was induced by suture method. Male SD rats (n=70) were randomly divided into sham operation group, I/R group, solvent control group, AS-IV group, AS-IV+autophagy inhibitor (3-methyladenine, 3-MA) group, 3-MA group and autophagy activator (rapamycin, Rapa) group. Except for sham operation group, the rats in other groups were subjected to ischemia for 2 h and reperfusion for 24 h. The rats with successful modeling were selected according to Zea Longa scoring criteria. The volume of cerebral infarction was measured by TTC staining. The morphological changes of nerve cells in the rats were observed with Nissl staining. The phenomenon of autophagy was observed under transmission electron microscope. The protein expression of beclin-1 and LC3-Ⅱ was determined by Western blot. RESULTS: No neurological deficit in sham operation group was observed, and the cerebral infarction was not found. Compared with sham operation group, obvious cerebral infarction was observed, the Nissl bodies were small in size and number and stained light, typical autophagosomes were observed, and the protein expression of beclin-1 and LC3-Ⅱ was increased in I/R group (P<0.05). Compared with I/R group, the volume of cerebral infarction was decreased obviously, neurological deficit restored significantly, and the number of autophagosomes and the protein expression of beclin-1 and LC3-Ⅱ were increased in AS-IV group and Rapa group (P<0.05). However, no significant difference between solvent control group and I/R group was observed (P>0.05). Compared with AS-IV group, the neurological deficit was serious, the volume of cerebral infarction and the number of autophagosomes were increased, while the expression of beclin-1 and LC3-Ⅱ was decreased in AS-IV+3-MA group and 3-MA group (P<0.05). CONCLUSION: Astragaloside IV may play an important role in atte-nuating cerebral ischemia/reperfusion injury by activating autophagy.     

Affecting factors of nylon monofilament induced focal cerebral ischemia in rats

AIM: To study factors affecting the stability of nylon monofilament for intraluminal middle cerebral artery occlusion in rats.METHODS:Successful rates and infarct volume of ischemic model were compared by polyvinyl alcohol (PVA) coated and silicon coated nylon monofilament intraluminal occlusion of middle cerebral artery under condition of ligation and without ligation of peterygopalatine artery.RESULTS:The successful rates were 60% and 53% in two groups under condition of ligation of peterygopalatine artery, and 20% and 27% in other two groups under condition of without ligation of peterygopalatine artery respectively. The intracranial length of nylon monofilament were about 7 mm in successful model and 4 mm in failure model. Animals in the PVA coated nylon monofilament group showed neurological dificit signs earlier, and had a significantly larger infarct volume at 12 hours of ischemia than those in the silicon coated group.CONCLUSIONS:The ligation of peterygopalatine artery is critical in the success of making this kind of ischemic model. The appropriate position of nylon monofilament entering crania and the good expansibility of PVA play an important role in occlusion of blood flow. Because of the relatively lower successful rate, a new substitute with unique quality, proper hardness and better elasticity needs to be explored in the future.     

Effects of baicalin on glial fibrillary acidic protein and nuclear factor-κB expression in juvenile rat hippocampus after status convulsion

AIM: To study the effects of baicalin (BC) on glial fibrillary acidic protein (GFAP) and nuclear factor-κB (NF-κB) expression and neuronal apoptosis in juvenile rat hippocampus after status convulsion (SC). METHODS: One hundred and ninety five juvenile male Sprague-Dawley rats were randomly divided into 3 groups: normal saline pretreatment group (NS group), SC group and SC with BC pretreatment group (BC group). Each of these 3 groups would be subdivided into 5 subgroups sacrificed at 4 h, 12 h, 24 h, 48 h and 72 h after SC. The rat SC model was prepared by lithium-pilocarpine chemical method. The protein expression of GFAP and NF-κB was detected by the method of immunohistochemistry. The mRNA expression of GFAP was detected by RT-PCR. The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). RESULTS: Compared with NS group, the GFAP positive cells was increased in SC group (P<0.05). Compared with SC group, the expression of GFAP was significantly reduced in BC group (P<0.05). Compared with NS group, the NF-κB positive cells was increased in SC group (P<0.05). Compared with SC group, the expression of NF-κB was significantly reduced in BC group. RT-PCR showed that the expression trend of GFAP mRNA was similar to that of the protein. Compared with NS group, the TUNEL positive cells in the hippocampus CA1 area in SC group increased significantly 12 h after SC (P<0.01), and reached a peak at 48 h. After the intervention with BC, the TUNEL positive cells decreased significantly between 12~48 h after SC (P<0.05 or P<0.01), but the number of TUNEL positive cells remained significantly greater than that in NS group (P<0.05). CONCLUSION: The expression of GFAP and NF-κB in the hippocampus increased after SC in rats. Baicalin decreases the expression of GFAP and NF-κB in hippocampus of rats with pilocarpine-induced seizures, and reduces the number of neuronal apoptosis, suggesting that baicalin may protect against the brain damage caused by status convulsion.     

Protective effect of atorvastatin on blood brain barrier in rats with focal cerebral ischemia-reperfusion injury

AIM: To explore the protective effects of atorvastatin on blood brain barrier(BBB) in cerebral ischemia-reperfusion(IR) injury and the potential mechanisms involved. METHODS: SD rats were divided into sham group, IR group and atorvastain group. Intraluminal suture method was used to establish cerebral IR model, and the ischemic brain was reperfused for 72 h after the occlusion. The rats in atorvastatin group were administered with atorvastatin(20 mg·kg^-1·d^-1) by gavage once a day for 3 consecutive days after operation. At 72 h after reperfusion, neurological function scores, the water content of the brain tissue, Evans blue(EB) content of ischemic hemisphere, the expression of tight junction(TJ)-associated protein occludin and inflammation factor phosphatidylinositiol 3-kinase-p110 gamma(PI3K-p110γ) were tested and analyzed. RESULTS: In IR group, the rats showed elevated neurological function scores(P<0.01), brain tissue water content(P<0.01) and EB content(P<0.01), accompanied with the down-regulation of occludin expression(P<0.01) and up-regulation of PI3K-p110γ(P<0.01) at 72 h after reperfusion. Compared with IR group, decreased brain edema(P<0.01) and EB leakage(P<0.01) were observed in atorvastatin group, accompanied with increased occludin expression(P<0.01) and decreased PI3K-p110γ expression(P<0.01). However, no statistical difference of the neurological function scores between the 2 groups was observed. CONCLUSION: Atorvastain attenuates cerebral IR injury, which may be associated with the inhibition of inflammatory reactions and the up-regulation of TJ-asso-ciated proteins to maintain the stability of BBB.     

Effects of bFGF on neuronal apoptosis and fractalkine expression in cerebral ischemia/reperfusion rats

AIM: To study the effects of basic fibroblast growth factor (bFGF) on neuronal apoptosis and fractalkine expression in ischemic penumbra after cerebral ischemia/reperfusion in rats.METHODS: Thirty-six rats were randomly divided into 3 groups: sham operation group, ischemia/reperfusion group and bFGF group. The model of middle cerebral artery occlusion was established by the method of intraluminal filament blockage. The middle cerebral arteries were blocked for 1 h and then reperfused for 24 h. Neurological performances of all rats were scored with Bederson's standard. The brain tissues of the rats were stained and the average infarct volume was calculated. TUNEL method was used to determine the number of apoptotic neurons, and the expression of fractalkine was detected by the method of immunohistochemistry.RESULTS: The score of neurological performances in bFGF group was 2.23±0.59, lower than that in ischemia/reperfusion group (3.18±0.65). The number of apoptotic neurons in bFGF group (13.22±1.35) was lower than that in ischemia/reperfusion group (17.28±1.01, P<0.05), which was the lowest in sham operation group (0.91±0.65). Compared with sham operation group, the expression of fractalkine in ischemia/reperfusion group was decreased. The expression of fractalkine in bFGF group was mainly higher than that in ischemia/reperfusion group (P<0.05).CONCLUSION: Up-regulation of fractalkine may be one of the molecular mechanisms of bFGF to protect neurons against ischemia/reperfusion injury.     

Combination of taurine and diazepam has neuroprotective effect on focal cerebral ischemia-reperfusion in rats

AIM: To observe the neuroprotective effect of combined treatment with taurine and diazepam against focal cerebral ischemia-reperfusion in rats. METHODS: Sixty male Sprague-Dawley rats were randomly divided into five groups: sham-operation group, vehicle group, taurine group (200 mg/kg, ip), diazepam group (10 mg/kg, ip) and combined treatment group (taurine 100 mg/kg+diazepam 5 mg/kg). Focal cerebral ischemia was induced by the method of middle cerebral artery occlusion (MCAO) in rats, and reperfusion was emerged by removing the thread 2 h later. The drugs were administered respectively at the time of reperfusion, and subsequently repeated once 12 h later. The animals in vehicle group were intraperitoneally injected with isodose normal saline. The neurological deficit score, the brain water content and cerebral infarction were measured 48 h after MCAO. Other 5 group animals of focal cerebral ischemia-reperfusion (n=16 in each group) were set up as mentioned above and accepted treatments 10 h after reperfusion, likewise repeated once 12 h later. Twelve animals in each group were adopted the same management as the previous 5 groups at 48 h after MCAO. The remained 4 animals in each group were sacrificed until two weeks after MCAO to observe the histopathological changes by nissl staining. RESULTS: Compared to vehicle group, the animals in combined treatment group at 2 h or 12 h after MCAO both decreased the neurological deficit score, reduced the brain water content and infarct volume (P<0.01 or P<0.05). The combined treatment significantly alleviated the neurological necrosis as well. The neuroprotective effect of the combined treatment was superior to that of using taurine or diazepam alone. CONCLUSION: These results suggest that combination of taurine and diazepam treatment has a coordinate neuroprotective effect on both the acute and chronic brain damage of focal cerebral ischemia-reperfusion.     

Effect of E-selectin pretreatment on cerebral ischemia-reperfusion injury in rats

AIM:To study the effect of nasal mucosal tolerance to E-selectin on cerebral ischemia-reperfusion injury.METHODS:Two different doses (single and booster) of E-selectin or PBS were dropped into membrana mucosa nasi of rats. The middle cerebral artery occlusion (MCAO) model referring to Zea Longa method with modifications was performed 48 h after the last dose of E-selectin or PBS. After 2 h ischemia and 22 h reperfusion, the numbers of CD3+CD4+T-lymphocyte and CD3+CD8+T lymphocyte subgroup in the blood were examined with flow cytometry. Rats were killed, then part of the animals was used to measure the cerebral infarction volume by TTC staining. mRNA expressions of E-selectin, ICAM-1 and lymphocyte function-associated antigen-1(LFA-1) were determined by RT-PCR and activity of SOD was determined by xanthinoxidanse method in ischemic cortex of the other part of animals. RESULTS:The ratio of the numbers of CD3+CD4+T-lymphocytes and CD3+CD8+T-lymphocytes increased in E-selectin single pretreatment group (P<0.05). Compared to other groups, E-selectin booster pretreatment group showed decreased CD3+CD8+T-lymphocytes (P<0.05), increased ratio of the numbers of CD3+CD4+T-lymphocytes and CD3+CD8+T-lymphocytes (P<0.05), reduced cerebral infarction volume by 40.87% (P<0.05), heightened activity of SOD (P<0.05), lowed E-selectin mRNA and ICAM-1 mRNA expression (P<0.05), and less tendency of LFA-1 mRNA expression.CONCLUSION:E-selectin induces cerebral ischemic tolerance and relieves cerebral ischemia-reperfusion injury. The mechanisms are related to the changes in the ratio of CD4+T-lymphocyte and CD8+T-lymphocyte. The heightened activity of SOD, the lowed mRNA expressions of E-selectin and ICAM-1, as well as the less tendency of LFA-1 mRNA expression are also involved.     

Effect of DADLE on lung injury in rats with acute global cerebral ischemia-reperfusion

AIM：To observe the effects of δ opioid receptor agonist DADLE on acute lung injury (ALI) induced by acute global cerebral ischemia-reperfusion in rats. METHODS：SD rats (n=30) were randomly divided into sham group, model (I/R) group and DADLE treatment group. Global cerebral ischemia-reperfusion model was established by a modified 2-vessel occlusion plus hypotension. DADLE (5 mg/kg) treatment was performed via the left jugular injection before reperfusion. After 120-min reperfusion, the pathological changes of the lung tissues were observed under light microscope and electronic microscope. The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) level were detected. The partial pressure of arterial oxygen (PaO2) was also measured. RESULTS：In I/R group, widened alveolar septum, capillary dilatation and congestion, endovascular and perivascular cells in the lung with neutrophil infiltration, and significantly reduced type II epithelial cell surface microvilli, alveolar lumen cavity and trachea with serous exudate were observed. SOD activity decreased, but the MDA level increased. Compared with I/R group, the SOD activity increased and MDA level decreased in DADLE treatment group, with significantly reduced lung congestion, the degree of lung injury, and the infiltration of neutrophils. Compared with I/R group, the PaO2 and oxygenation index in DADLE treatment group were increased. CONCLUSION：Various degrees of pulmonary injury were observed in acute global cerebral ischemia reperfusion model. DADLE might have a protective effect on lung tissues of ALI in rats.     

Effect of electroacupuncture on astrocyte morphology of ischemia cortex penumbra zone in cerebral infarct rats

AIM: Effect of electron acupuncture on astrocyte morphology of cerebral ischemic penumbra zone in rats was observed.METHODS: The brain infarct model in male Sprague-Dawley rats was made by thread embolism cerebral middle artery.Immunochemistry method was used to observe the change of astrocyte morphology in cerebral ischemic penumbrazone.RESULTS: There was no statistically difference in astrocyte morphology between cerebral infarct group and electroacupuncture treatment group.CONCLUSION: Acupoint stimulation of 5 and 10 Hz electro-acupuncture with a current intensity of 2 mA may not affect on astrocytic morphology.     

Changes of cortisol content in plasma and hippocampus after focal cerebral ischemia-reperfusion in rats

AIM: To investigate the relationship between glucocorticoid (Gc) and injury of hippocampus neurons and the effect of Gc on dementia episode after cerebral ischemia-reperfusion. METHODS: The rat model of middle cerebral artery occlusion (MACO) was established. Cortisol contents in hippocampus and plasma of the model rats were examined by means of the radioimmunoassay at 2 h, 6 h, 12 h, 24 h after reperfusion. RESULTS: The levels of cortisol content in model group were significantly higher than those in sham group and normal group both in hippocampus and plasma. The highest cortisol content was observed at 6 hours after reperfusion. HE staining showed that the impairment of hippocampus neurons was aggravated progressively with reperfusion interval elongating. CONCLUSION: The increased cortisol in hippocampus and plasma, after 2 h cerebral ischemia and 24 h reperfusion, could aggravate the injury of hippocampus neurons and lead to dementia post stroke.     

Inhibitory effect of ischemic postconditioning on autophagy induced by focal cerebral ischemia reperfusion in rats

AIM: To investigate the effect of ischemic postconditioning (IPC) on autophagy induced by focal cerebral ischemia reperfusion (I/R) in rats. METHODS: Healthy male SD rats were assigned randomly into sham-operation (sham) group, I/R group and IPC group with 10 rats in each group. The rats in sham group were only exposed the right common, internal and external carotid artery surgically. The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion. The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h. Autophagy was obeserved by transmission electron microscopy (TEM). The protein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot. Pathological changes of brain tissue were observed by HE staining. RESULTS: The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05). The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01). The cerebral infarction area and brain water content in IPC group were significantly lower than those in I/R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I/R group. TEM observation showed that IPC revealed fewer autophagosomes, with much less severe cell damage than that in I/R group. CONCLUSION: IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells, which might be related to the activation of mTOR.     

Effect of genistein on ammonia-induced NF-κB activation in astrocytes

AIM: To explore the effect of genistein on ammonia-induced nuclear factor-κB (NF-κB) activation and the underlying mechanism.METHODS: Primary astrocyte cultures were prepared and challenged with NH₄Cl to establish a hyperammonemic model. The activation of ERK, Akt and NF-κB was examined by Western blot.RESULTS: AG1478 and genistein significantly inhibited ammonia-induced activation of ERK and Akt. Ammonia-induced NF-κB nuclear translocation was significantly inhibited by the pretreatment of LY294002, genistein and AG1478.CONCLUSION: Genistein significantly inhibited ammonia-induced ERK activation and Akt-mediated NF-κB activation, which might represent the important mechanism by which this naturally occurring substance exerts its swelling-inhibiting effect.     

Effect of sedation by infusion of dexmedetomidine on inflammatory factors in patients with severe trauma

AIM: To study the effect of short-term sedation by infusion of dexmedetomidine (DEX) on the inflammatory factors in the patients with severe trauma. METHODS: Sixty patients with severe trauma in ICU were randomly divided into 3 groups: DEX group (n=20), midazolam (MDZ) group (n=20) and control group (without any sedatives, n=20). Dexmedetomidine at an initial loading dose of 1~2 μg/kg was administered intravenously over 10 min to the patients in DEX group before the sedation procedure of continuous infusion at dose of 0.2~0.7 μg·kg^-1·h^-1 by a 50-mL infusion syringe was perform. Midazolam at dose of 0.03~0.3 mg/kg was also administered intravenously immediately before the procedure of a continuous infusion at dose of 0.03~0.2 mg·kg^-1·h^-1. The doses of DEX or MDZ for the patients receiving short-term (48 h) sedation were adjusted according to the Ramsay sedation scale scores. The serum levels of interleukins (IL-1 and IL-6), tumor necrosis factor α (TNF-α) and C-reactive protein (CRP) were measured at the time points of the baseline (before the start of the study), 24 h and 48 h of sedative infusion. RESULTS: The levels of TNF-α, IL-1, IL-6 and CRP in the 3 groups of severe traumatic patients at the time when they were admitted to the hospital were significantly higher than those of the normal values. The levels of TNF-α, IL-1, IL-6 and CRP in control group gradually increased at 24 h and 48 h, while those in DEX group and MDZ group significantly declined. No significant difference of TNF-α, IL-1, IL-6 and CRP levels between DEX group and MDZ group was observed at 24 h, but the changes were significantly different at 48 h. CONCLUSION: Dexmedetomidine reduces over-stress responses to a certain extent in the traumatic patients to prevent the further production and release of inflammatory mediators, thus contributing to the stability and recovery of the patients with severe trauma.