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Objective:To explore the effect of SimMan 3G simulator based scenario simulation teaching method and case-based learning (CBL) in emergency medicine teaching.Methods:Sixty students from Batch 2013 eight-year program of clinical medicine were selected as subjects. They were randomly divided into an experimental group and a control group, with 30 students in each group. In the teaching of emergency medicine, the experimental group used the combination of scenario simulation with CBL teaching methods, and the control group used classic teaching methods. The test scores and the questionnaires satisfaction of the two groups were compared to evaluate the teaching effects. SPSS 17.0 was used for the statistical analysis, measurement data were compared between the groups by t test, and counting data were compared between groups by chi-square test.Results:The scores of the experimental group (94.24±1.13) were better than those of the control group (90.6±0.59), with significant differences ( t=12.85, P<0.05). The results of the questionnaires showed that the students of experimental group were more satisfied with the learning experience than those of the control group. Conclusion:The teaching method can improve the teaching effects, the students' emergency clinical thinking, skills, comprehensive analysis and judgment ability, team cooperation consciousness and leadership ability.
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Objective To investigate the effects of exogenous hydrogen sulfide (H2S) on injury of rat hippocampus neurons induced by oxygen-glucose deprivation and restoration (OGD/R) and explore its mechanism.Methods Hippocampus neurons were isolated from embryonic day 16-18 (E16-18) rat embryos.Hippocampus was immediately removed and digested with 0.25％ trypsin.The neurons were isolated and cultured at 37 ℃ for 7 days and neuron-specific enolase (NSE) was detected by immunohistochemical staining method to identify neurons.At 8th day,the neurons were placed in deoxygenated glucose-free medium and exposed to 95％ N2-5％ CO2 in an air tight chamber for 1 hour,and then replaced the glucose-free medium with original medium,and returned the cultures to a standard incubator in 5％ CO2 at 37 ℃ and incubated for another 24 h.The neurons were divided into 3 groups:group Ⅰ control; group Ⅱ OGD/R,and group ⅢOGD + NaHS,the latter was further divided into 5 subgroups:groups Ⅲ1-5 with 25,50,100,200,400 μmol/L NaHS added,respectively.Then cell viability was quantified by MTT method,the level of lactate dehydrogenase (LDH) were detected,apoptosis was measured by Annexin V FITC/PI Apoptosis Kits,and RT-PCR was used to assay HIF-1 α mRNA in neurons in all groups.Results Compared with control group,the LDH level,apoptosis and expression of HIF-1α mRNA in group Ⅱ were significantly increased,the cell viability was significantly decreased (P ＜ 0.01).There were no significant differences in the LDH level,apoptosis and expression of HIF-1 α mRNA and the cell viability between group Ⅱ and group Ⅲ1 (P ＞ 0.05).Compared with group Ⅱ,the LDH level,apoptosis and expression of HIF-1α mRNA in group Ⅲ2-4 were significantly increased,the cell viability was significantly increased (P ＜ 0.01).Compared with group Ⅱ,the LDH level,apoptosis and expression of HIF-1 α mRNA in the group Ⅲ 5 were significantly decreased,the cell viability was significantly decreased (P ＜ 0.01).Conclusions H2S of low concentration has no significant effects on injury of rat hippocampus neurons induced by OGD/R.H2S of moderate doses can protect rat hippocampus neurons from OGD/R injury and H2S of high concentration can aggravate injury.The expression of HIF-1α mRNA in rat hippocampus neurons was increased after OGD/R,and the protective role of H2S is associated with increase in the expression of HIF-1α mRNA.
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Objective To explore the effects of hydrogen sulfide (H2S) on apoptosis of cardiomyocytes after cardiopulmonary resuscitation (CPR) in rat models.Methods Forty-five male SD rats were randomly into sham group (n =15),CPR group (n =15) and NaHS group (n =15).Rats of CPR group and NaHS group were operated to induce cardiac arrest by transcutaneous electrical stimulation to epicardium.In NaHS group,NaHS (5 mg/kg) was administrated via the femoral venous line 1 min before CPR.Hemodynamic variables were monitored and obtained continuously.Survival rats were sacrificed at 24 h after restoration of spontaneous circulation and the hearts were removed for analysis by RT-PCR and TUNEL assays.Blood samples were collected and plasma content of cTnT was detected.Results Compared with the CPR group,animals treated with NaHS had improved left ventricular function (P ＜0.01),lower plasma cTnT levels (P ＜0.05) and decreased apoptosis index (P ＜ 0.01) 24 h after ROSC.The expressions of Caspase-3 mRNA,Bax mRNA and Bcl-2 mRNA in CPR group and NaHS group were higher compared with the control group (P ＜0.01).The NaHS group had lower expressions of Caspase-3 mRNA and Bax mRNA (P ＜0.01),but higher expression of Bcl-2 mRNA (P ＜0.05) compared with the CPR group.Conclusions Exogenous (H2S) regulated the expressions of Caspase-3,Bax and Bcl-2 mRNA,thereby preventing apoptosis of cardiomyocytes,inhibiting cTnT release and improving left ventricular function 24 h after CPR.
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Objective To study the efficacy of transurethral holmium laser resection as well as instant intravesical Pirarubicin instillation for superficial bladder carcinoma.Methods 51 cases with superficial bladder cancer were divided into observation group(A) including 26 cases and control group(B) including 25 cases stochastically.The 26 cases of group A were received transurethral holmium laser resection as well as instant intravesical Pirarubicin instillation therapy whereas 25 cases of group B received transurethral resection therapy.All patients of both groups were treated with intravesical instillation of Pirarubicin therapy regularly at least 1 year and have been followed up for 15 to 39 months.Results Differences were not significant between the two groups in the mean operation time and bladder perforation rate(P ＞0.05 ),but indwelling period of urethral catheter was markedly decreased in group A than group B ( P ＜ 0.05 ).There were 4 cases of recurrence in group A,recurrence rate was 15.4％ (4/26).While there were 11 cases of recurrence in group B,recurrence rate was 44.0％ (11/25 ).Significant difference in cancer recurrence rate was found between two groups ( P ＜ 0.05 ).Conclusion Compared with transurethral resection therapy transurethral holmium laser resection as well as instant intravesical Pirarubicin instillation for superficial bladder carcinoma was more effective and quicker recovery,which has a good applied future in the clinical practice.
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Objective To investigate the effects of hydrogen sulfide on sepsis-induced cardiac dysfunction in rats.Methods Male SD rats were randomly ( random number) divided into 4 groups:control group ( Ⅰ group),sepsis group ( Ⅱ group),sepsis + NaHS group (Ⅲ group),sepsis + PAG group (Ⅳ group). Cecal ligation and puncture technique was used in Ⅱ, Ⅲ, Ⅳ group. Hemodynamic was observed,and H2S synthesis,CSE (cystathionine-r-lyase) mRNA,MPO activity and the level of TNF-α,IL-1β were determined.The morphological changes and infiltration of inflammatory cells in myocardium were also observed.Results Compared with Ⅰ group,H2S synthesis,the levels of TNF-α,IL-1 β,the activity of MPO increased ( P ＜ 0.05 or P ＜ 0.01 ),and the expression of CSE-mRNA increased,and blood pressure of rats decreased significantly in Ⅱ group.Compared with Ⅱ group,H2S synthesis,the levels of TNF-α,IL-1β and the activity of MPO increased (P ＜ 0.05),the expression of CSE mRNA did not change noticeably ( P ＞ 0.05),and blood pressure of rats decreased more significantly in Ⅲ group.Compared with Ⅱ group, H2S level decreased,the levels of TNF-α,IL-1β and the activity of MPO decreased significantly,the expression of CSE mRNA decreased and blood pressure of rats decreased in Ⅵ group (P ＜0.05).Histopathological changes of myocardium were aggravated in the following severity order: Ⅰ ＜ Ⅳ ＜＜ Ⅲ.Conclusions CSE/H2S system of the myocardium was upregulated in sepsis rats.Hydrogen sulfide could raise the levels of MPO,TNF-α,IL-1β,aggravating myocardial injury. Contrarily,the inhibitor of H2S could counteract it.
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To investigate the interaction and involvement of sodium hydrosulfide (NaHS), a H(2)S donor, on hippocampus of rats suffering from sepsis-associated encephalopathy, rats were subjected to cecal ligation and puncture (CLP)-induced sepsis. Adult male Sprague-Dawley rats were randomly divided into four groups: Sham group, CLP group, CLP+NaHS group and CLP+aminooxyacetic acid (AOAA, an inhibitor of H(2)S formation) group. The four groups were observed at 3, 6, 9, 12 h after treatment. We examined hippocampal H(2)S synthesis and the expression of cystathionine-β-synthetase (CBS), a major enzyme involved in the H(2)S synthesis in hippocampus. CBS expression was detected by reverse transcription polymerase chain reaction (RT-PCR). The concentrations of inflammatory cytokines (TNF-α, IL-1β) were determined in hippocampus by using enzyme-linked immunosorbent assay (ELISA). Neuronal damage was studied by histological examination of hippocampus. In CLP group, H(2)S synthesis was significantly increased in hippocampus compared with sham group and it peaked 3 h after CLP (P<0.05). Sepsis also resulted in a significantly upregulated CBS mRNA in hippocampus. The levels of TNF-α and IL-1β in the hippocampus were substantially elevated at each time point of measurement (P<0.05), and they also reached a peak value at about 3 h. Administration of NaHS significantly aggravated sepsis-associated hippocampus inflammation, as evidenced by TNF-α and IL-1β activity and histological changes in hippocampus. In septic rats pretreated with AOAA, sepsis-associated hippocampus inflammation was reduced. It is concluded that the rats subjected to sepsis may suffer from brain injury and elevated pro-inflammatory cytokines are responsible for the process. Furthermore, administration of H(2)S can increase injurious effects and treatment with AOAA can protect the brain from injury.
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Objective To investigate the changes of the expression of HIF-1α (hypoxia induciblefactor-1a) mRNA in myocardium of rats after cardiopulmonary resuscitation (CPR) and the intervention effects of exogenous hydrogen sulfide on it. Method Forty five male SD rats were randomly divided into three groups: control group ( n = 15 ), cardiopulmonary resuscitation group ( CPR group, n = 15 ) and NaHS + CPR group (n = 15 ) . Rats of control group were anesthetized and intubated without asphyxia and cardiac arrest. The rats of CPR group and NaHS + CPR group were operated to induce cardiac arrest by asphyxiation. In the rats of NaHS + CPR group, NaHS in dose of 50 ug/kg was administrated via the femoral venous line 1 minute before CPR. Hemodynamic was monitored continuously. The expression of HIF - lα mRNA in myocardium of rats in each group was determined by using RT-PCR and the activity of myeloperoxidase (MPO) in the myocardium of rats in each group was assayed by using a patent reagent box 6 h after CPR. The histopathological changes of myocardium were also observed. The t- test was used for statistical analysis. Results There was no statistically significant difference in hemodynamic changes between CPR group and CPR + NaHS group ( P ＞ 0. 05 ) . When compared with the control group, the activity of MPO and the expression of HIF-1α mRNA in CPR group and CPR + NaHS group were both increased, and those increased in CPR + NaHS group was more significant (P ＜ 0. 05) . When compared with CPR group, the expression of HIF-1α mRNA in CPR + NaHS group was higher, however, the activity of MPO in CPR + NaHS group was lower ( P ＜ 0. 05) . There were various histopathological changes of myocardium of rats found in CPR group and CPR + NaHS group, and the damage of myocardium of rats in CPR group was more obvious than that in CPR + NaHS group. Conclusions The expression of HIF-1α mRNA in myocardium of rats was increased after CPR. Exogenous hydrogen sulfide can protect myocardium cells from resuscitation-perfusion injury, and the protection is associated with increase in expression of HIF 1 αmRNA.
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Objective To determinate the effects of sodium tanshinone ⅡA sulfonate(STS)on cardiomyocyte hypertrophy and explore the relative effects of STS on mitogen-activated protein kinase signal transduction system in rats with cardiomyocyte hypertrophy through constricting the thoracic aorta.Methods The models of cardiomyocyte hypertrophy were established in vivo,and the thoracic aorta was partially tied between the right innominate and the left common carotid arteries.The rats were randomly divided into 6 groups(n=8/group)as follows:①sham,②transverse aortic constriction(TAC),③TAC+low-dose Tan(TAC+LT)(5 mg/kg),④TAC+middle-dose Tan(TAC+MT)(10 mg/kg),⑤TAC+high-dose Tan(TAC+HT)(20 mg/kg),and ⑥ TAC+Val(10 mg/kg).After treatment for 8 weeks,echocardiography was performed to observe the changes in hypertrophy and heart function,and heart samples were cut into transverse sections and stained with hematoxylin and eosin(H&E).The MAPKs protein expression in the cardiomyocytes was detected by Western blot.Results The heart weight index(HWI),left ventricular mass index(LVMI)and cross-sectional diameter of cardiomyocytes(CD),left ventricular posterior wall thickness(LVWT),and interventricular septal thickness(IVS)were significantly increased in TAC group as compared with sham group.The relative parameters in STS groups and Val group were reduced as compared with those in TAC group.Western blot analysis revealed the p-ERK and p-p38 expression was significantly decreased in TAC group as compared with sham group(P＜0.01).The p-ERK expression was significantly decreased in STS groups and Val group as compared with TAC group(P＜0.05).The TAC+HT group,TAC+MT group and Val group had significantly higher p-p38 expression than TAC group(P＜0.05).Conclusion Tanshinone ⅡA could regulate the expression of protein in MAPK pathway to exert its inhibitory effects on hypertrophy of cardiomyocytes.
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Objective To investigate the protective effects of endotoxin precondition on hepatic tissue in rats with endotoxemia.Method The models of rats with acute endotoxemia were produced by injecting LPS directly.Seventy-two male wistar rats were randomly divided into three groups:saline control group(N,n=24),lipopolysaccharide (LPS)-treated group(L,n=24),LPS pretreated group(P,n=24).Each group was divid-ed into four subgroups:saline control and lipopolysaccharide (LPS)-treated 2 h,4 h,6 h,12 h groups and LPS-pretreated 2 h,4 h,6 h,12 h groups.Rats in group P were first administered with introperitoneal injection of 0.25 mg/kg LPS,and after 24 hours,the rats were injected with 0.5 mg/kg LPS.Rats in group N and group L received with an equivalent amount of saline.After 72 hours,rats in group L and group P were intravenonsly injected with 10 mg/kg LPS,and rats in group N received with an equivalent amount of saline.Six rats were killed at 2,4,6 and 12 hours after injection of LPS in group L and P.The hvers were removed for detecting Toll like receptor-4 (TLR-4),nuclear factor-кB(NF-кB),tumor Necrosis Factor-apha(TNF-α)and malondialdehyde(MDA).The blood was drawn for detecting Alamine aminotmnsferose (ALT) and Aspartate aminotransferose (AST).The patho-logical changes of liver were also examined.Software SPSS13.0 was utilized to do ANOVA for statistical analysis.Results The rats exposed to LPS alone demonstrated an increase in TLR-4.NF-кB and TNF-α activity of the liver tissue.Incontrast.the rats exporsed 10 LPS prelreatment exhibited a significant decrease in TLT-4,NF-кB and TNF-α activity.The contents of TLR-4,NF-кB and TNF-α of LPS-treated 4 h groupwere,(38.76±0.67),170.82 ±31.40),293.16±49.49)and(6.263±0.351),significantly higher than those of the saline control group.The administration of endotoxin pretreatment reduced the indexes to(22.35±1.35),(135.55±26.44)and(234.23±44.96),respectively(P＜0.05).Conclusions TLR-4,NF-кB and TNF-α take part in the progress of hepatic injury in rats with endotoxemia.Endotoxin pretreatment can eliminate hepatic injury and protect the hepatic tissue by downmgulating the levels of TLR-4.NF-кB and TNF-α.
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No abstract available.
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Intestinal ischemia-reperfusion (I/R) is an important event in the pathogenesis of multiple organ dysfunction syndrome (MODS). The aim of this study is to determine the effects of ginsenoside Rb1 on liver injury induced by intestinal I/R in rats. Adult male Wistar rats were randomly divided into four groups: (1) a control, sham-operated group (sham group); (2) an intestinal I/R group subjected to 1 h intestinal ischemia and 2 h reperfusion (I/R group); (3) a group treated with 20 mg/kg ginsenoside Rb1 before reperfusion (Rb1-20 group); and (4) a group treated with 40 mg/kg ginsenoside Rb1 before reperfusion (Rb1-40 group). Liver and intestinal histology was observed. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) level in serum and malondialdehyde (MDA) level in intestinal tissues were measured. Myeloperoxidase (MPO), TNF-alpha, MDA level and immunohistochemical expression of NF-kappa B and intracellular adhesion molecale-1 (ICAM-1) in liver tissues was assayed. In addition, a western blot analysis of liver NF-kappa B expression was performed. Results indicated intestinal I/R induced intestinal and liver injury, which was characterized by increase of AST and ALT in serum, MDA level in intestine, MPO, TNF-alpha and MDA level and ICAM-1 and NF-kappa B expression in the liver tissues. Ginsenoside Rb1 (20, 40 mg/kg) ameliorated liver injury, decreased MPO, TNF-alpha and MDA level, NF-kappa B and ICAM-1 expression in liver tissues. In conclusion, ginsenoside Rb1 ablated liver injury induced by intestinal I/R by inhibiting NF-kappaB activation.
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Animals , Male , Rats , Alanine Transaminase/blood , Aspartate Aminotransferases/blood , Biomarkers/metabolism , Ginsenosides/pharmacology , Intercellular Adhesion Molecule-1/metabolism , Intestines/blood supply , Ischemia/metabolism , Liver/enzymology , Liver Diseases/etiology , Malondialdehyde/metabolism , NF-kappa B/antagonists & inhibitors , Peroxidase/metabolism , Rats, Wistar , Reperfusion Injury/complications , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Background Left ventricular hypertrophy(LVH) is a cardiovascular risk factor independent of the blood pressure. JAK/STAT pathway has been confirmed to participate in cardiac hypertrophy and hyperplasia. Our previous reports have shown that sodium tanshinone ⅡA sulfonate(STS) reversed LVH,inhibited the myocardial cells Ca2+ influx,lowered left ventricular myocardial tumor necrosis factor-?(TNF-?)and the proto-oncogene c-fos,Bcl-2,and p53 protein expression. Objective To study the effect of sodium tanshinone ⅡA sulfonate(STS) on JAK/STAT pathway in left ventricular hypertrophy(LVH) induced by abdominal aorta stenosis in rats. Methods Twenty-four 9-weeks-old rats submitted to abdominal aorta constriction,were randomized to receive STS 10 mg/(kg?d)(n=8)or sterilized distilled water (1 mL/d)(n=8),or valsartan 10 mg/(kg?d) by gavage(n=8),with age and sex matched sham operated rats(n=8) as control. HE,VG and immunohistonchemical staining were used to evaluate the myocardial fiber dimension(MFD). Expressions of JAK1 and STAT3 were assessed by using Western blot. Results Compared with the control group,pressure loaded rats had higher SBP[(117.3?8.3) vs LVH: (186.5?13.5)mmHg,P
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Objective: To observe the changes of hemeoxygenase-1 (HO-1) activity in cerebral tissue, carboxyhemoglobin (COHb) level in blood of rats during global cerebral ischemia/reperfusion (I/R) and the influence of L-tetrahydropalmatine (L-THP) on them. Methods: Seventy-seven Sprague-Dawley (SD) rats were randomly divided into sham-operated group (S group, n=7), I/R group (I group, n=35) and L-2THP treatment group (T group, n=35). Cerebral I/R model was reproduced in SD rats of I group and T group. HO-1 activity, cyclic GMP (cGMP) in the brain and COHb in blood were evaluated respectively at 1, 3, 12, 24, and 48 hours after global cerebral I/R and compared to those of the S group. Results: ① HOCD*21 activity, COHb content and cGMP level in I group increased as compared with those in S group after global cerebral I/R (all P
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BACKGROUND: Being a traditional Chinese herb, huangqi (astragalus membranaceus) resists free radical, enhances immunity, promotes microcirculation and protects vascular endothelial cell and nervous system; of which, the protection of huangqi (astragalus membranaceus) on nervous vessel is related to its regulation on local cerebral cortical blood flow rate. OBJECTIVE: To observe the effects of huangqi (astragalus membranaceus) on local cerebral cortical blood flow rate after traumatic injury. DESIGN: Randomized paired experiment.SETTING: Neurological Surgical Institute of Lanzhou Military Area Command of Chinese PLA.MATERIALS: The experiment was performed in Traumatic Experimental Room of Neurological Surgical Institute of Lanzhou Military Area Command of Chinese PLA from February to May 2000, in which, 75 healthy male SD rats were employed and randomized into the control (5 rats), saline control (35 rats) and experimental group (35 rats). The latter two groups were subdivided into 7 time spots of 1, 3, 6, 8, 12, 24 and 48 hours after traumatic injury, 5 rats were involved in each spot.METHODS: In normal control, the model was not prepared and in the other two groups, the modified Feeney's method was used to establish craniocerebral traumatic model. In experimental group, right after injury, huangqi injection (5 g/kg) was applied abdominally and in saline control, physiological saline 0.5 mL was injected abdominally. MAIN OUTCOME MEASURES: It was to determine the blood flow rates in local cerebral cortex at corresponding time spots after traumatic injury. Fenestration was done in normal control for direct determination. RESULTS: By supplemented, 75 rats entered result analysis. Blood flow rates in local cerebral cortex: the rate in saline control 1 hour after trauma was lower than the control [(6.90±0.68), (7.94±0.65), P < 0.05], it was de creased to the minimum in 24 hours and began increasing in 48 hours [(5.86±0.61), (6.15±0.60)]; the rate in experimental group at every time spot was higher than saline control (P < 0.05). CONCLUSION: Huangqi (astragalus membranaceus) increases remark ably blood flow rate in local cerebral cortex, which is associated with its neurovascular protection.
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To investigate the role of NF-κB in endotoxic shock in rats, the model of endotoxinshock rats was induced by intravenous infusion of lipopolysaccharide (LPS). 1 h, 2 h, 4 h and 6h after LPS injection, the activation of NF-κB in blood mononuclear cells and the content of TNF-α and IL-6 in plasma was detected by enzyme-linked immunoadsordent assay (ELISA). The level of mean arterial pressure (MAP) and the histopathological changes of lung and liver were also observed. The activation of NF-κB in mononuclear cells increased 1 h after LPS injection and reached its peak 2 h after the injection, and its level was higher than that of normal group. The level of TNF-α was increased 1 h after the infusion and peaked 2 h after the injection, and its level was higher than that of normal group after LPS infusion. The content of IL-6 increased gradually with time, the IL-6 level was higher than that of normal group after LPS injection. MAP was decreased gradually with time and its level was lower than that of normal group after LPS injection. Pathological examination showed that endotoxic shock could cause pulmonary alveolar hemorrhage, edema and infiltration of inflammatory cell in lung tissue and congestion, edema, capillary dilation and inflammatory cell infiltration in liver tissue. It is concluded that NF-κB can up-regulate the expression of TNF-α and IL-6 in plasma and play an important role in endotoxin-induced shock in rats.
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BACKGROUND: The studies in recent years proved that the inflammatory reaction is of the main reasons in the damage of cerebral ischemia reperfusion. The nuclear factor κB (NF-κB), as a kind of transcription factor, plays an important role in regulating the expressions of various inflammatory cell factors in the inflammatory reaction of cerebral ischemia reperfusion. The previous experiments show that puerarin functions to resist the oxidated free radicals and the apoptosis of nerve cells. In case it has the functions of anti-inflammation, its brain protection can be explained further.OBJECTIVE: To study the effect of puerarin on NF-κB for the rats with the damage of ischemia reperfusion.DESIGN: A random parallel controlled study.SETTING: The Emergency Department of Beijing Hospital, Emergency Department of Tongji Hospital, Pathology Department and Experimental Animal Center of Tongji Medical College, and Health Statistics Department of Public Health College of Huazhong University of Science and Technology.MATERIALS: The experiment was started on April 12, 2003 in the Pathology Department of Tongji Medical College. The 75 healthy and clean Wistar rats were randomized into 3 groups with 25 in each, Sham operation group, cerebral ischemia reperfusion group, and puerarin group. Each group was reperfused at 2, 6, 12, 24, and 48 hours after ischemia and 5rats were used at each time point.METHODS: [1] Sham operation group: Without electric coagulation of bilateral vertebral arteries, without blockage of bilateral common carotid arteries, without medicinal administration. [2] Cerebral ischemia reperfusion group: Ten minutes after the blockage of bilateral common carotid arteries with non-invasive artery clamp, the reperfusion was given. At the beginning of reperfusion, the abdominal injection of normal saline 1 mL was applied and later every 6 hours the injection was repeated once. [3] Puerarin group:The procedure was the same as for the reperfusion group, only with normal saline changed to puerarin 100 mg/kg.MAIN OUTCOME MEASURES: At the time points of 2, 6, 12, 24, and 48 hours after reperfusion, the activity of NF-κB and inhibitory protein κB(IP-κB) in the hippocampus CA1 region was examined with immunohistochemical method; the expression of tumor necrosis factor-α (TNF-α) mRNA was measured with in situ hybridization method; and the number of surviving neurons was detected with hematoxylin-eosin (HE) staining.RESULTS: After supplement, 75 rats entered the result analysis. [1] Activity of NF-κB: In the ischemia reperfusion group, it was obviously increased at 2 hours after reperfusion, to the highest at 6 hours, and still higher than that of the sham operation group, (P ＜ 0.01). In the puerarin group, it was lower at each time point than that of the ischemia reperfusion group (P ＜ 0.01). [2] Expression of TNF-α mRNA: In the ischemia reperfusion group, it was obviously increased at 2 hours after reperfusion, to the highest at 12 hours, and still higher than that of the sham operation group at 48 hours (P ＜ 0.01). In the puerarin group, it was lower than that of the ischemia reperfusion group at 6-48 hours (P ＜ 0.01). [3] Activity of IP-κB:In the ischemia reperfusion group, it was obviously decreased at 2 hours after reperfusion, to the lowest at 6 hours, and then gradually increased to the level of 12 hours. In the puerarin group, it was higher than that of the ischemia reperfusion group at each time point (P ＜ 0.01 or 0.05). [4] Number of surviving neurons: In the ischemia reperfusion group, it was decreased gradually with the time prolonging after reperfusion (P ＜ 0.01). In the puerarin group, at each time point, it was higher than that of the ischemia reperfusion group (P ＜ 0.05 or 0.01).CONCLUSION: In the cerebral ischemia reperfusion, puerarin can protect the brain through decreasing the degradation of IP-κB, the activity of NF-κB, the expression of TNF-α mRNA, and the inflammatory reaction.
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To investigate the effects of L-Tetrahydropalmatine (L-THP) on the expressions of bcl-2, bax and neuronal apoptosis after cerebral ischemia and reperfusion, 60 Wistars rats were randomly divided into 3 groups: sham-operation group (group S, n = 20), ischemic-reperfusion group treated with saline (group I, n = 20) and ischemia-reperfusion group treated with L-THP (group T, n = 20). The rat model of global cerebral ischemia and reperfusion was induced by Pulsinelli's four-vessel occlusion method. The expression of bcl-2 and bax mRNA was detected by in situ hybridization and reverse transcriptional polymerase chain reaction (RT-PCR). The number of apoptotic neurons was examined by terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. Compared with group S, the expression of bcl-2 and bax mRNA in group I was increased significantly (P<0.01), and the number of apoptotic neurons increased either (P< 0.01). After L-THP treatment, the expression of bcl-2 mRNA was up-regulated (P< 0.01) and that of bax mRNA was down-regulated (P<0.01); the number of apoptotic neurons was decreased (P<0.01). Our results indicated that bcl-2 may suppress apoptosis and bax promote apoptosis after cerebral ischemia and reperfusion. L-THP could ameliorate cerebral ischemia and reperfusion damage by reducing the apoptosis through regulating bcl-2 and bax.
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Animals , Rats , Apoptosis , Berberine Alkaloids , Pharmacology , Brain Ischemia , Metabolism , Pathology , Neurons , Pathology , Proto-Oncogene Proteins c-bcl-2 , Genetics , RNA, Messenger , Genetics , Random Allocation , Rats, Wistar , Reperfusion Injury , Metabolism , Pathology , bcl-2-Associated X Protein , GeneticsABSTRACT
AIM To study the effects of puerarin on the expression of Bcl-2 and Bax, genes relating to neuronal apoptosis after cerebral ischemia and reperfusion. METHODS After global cerebral ischemia followed by reperfusion, changes in protein expression of Bcl-2 and Bax were detected by immunohistochemical method. The number of neuronal apoptosis was assessed by TUNEL. The effects of puerarin intervention were observed. RESULTS In CA1, the level of positive expression of Bcl-2 varied to the duration of reperfusion and the peak level was at 6 h reperfusion after 10 min global cerebral ischemia,the peak expression of Bax was at 24 h. The number of neuronal apoptosis after cerebral ischemia reperfusion was increased. In puerarin group, the expression of Bcl-2 was up-regulated and that of Bax was down-regulated, the number of neuronal apoptosis after cerebral ischemia reperfusion was decreased. CONCLUSION Our result indicate that Bcl-2 may restrain apoptosisl. Bax may promote apoptosis after cerebral ischemia and reperfusion and puerarin ameliorate ischemic damage by reducing the apoptosis through regulating Bcl-2 and Bax.