Influence of Electroacupuncture on the mRNA of Heat Shock Protein 70 and 90 in Brain after Cerebral Ischemia/Reperfusion of Rats

To explore the anti-apoptotic role of electroacupuncture (EA) and its molecular mecha-nisms after cerebral ischemia/reperfusion (IR) of rats, by using animal model of middle cerebralartery occlusion (MCAO), the changes of the cleavage of PARP were observed by Western blot andthe mRNA of heat shock protein (Hsp) 70 and Hsp90β detected by competitive RT-PCR after cere-bral IR and EA treatment. The results were as follows: (1) The cleavage of PARP was increased inischemic hippocampus, and EA treatment could attenuate the level of the cleavage remarkably; (2)The mRNA expression of Hsp70 was increased in the ischemic cortex and hippocampus and was fur-ther increased after EA treatment; (3) The mRNA expression of Hsp90β was decreased in ischemiccortex and hippocampus and the decrease was relatively slight after EA treatment. The above resultsdemonstrated EA treatment could protect neurons from apoptosis after cerebral IR. One of the molec-ular mechanisms was the promotion of the inducible expression of Hsp7     

Dynamic expression of glucose transporters 1 and 3 in the brain of diabetic rats with cerebral ischemia reperfusion

Background Blood glucose control improves the outcome of diabetic patients with stroke, but the target range of blood glucose control remains controversial. The functional recruitment of ischemia penumbra is extremely important to the recovery after stroke. The present study aimed to explore the expression of brain-type glucose transporters （GLUT1 and GLUT3） in cerebral ischemic penumbra at different blood glucose levels and different ischemic-reperfusion time in diabetic hypoxia-ischemia rats. The results might provide an experimental basis for clinical treatment of diabetic patients with stroke. Methods The Wistar rats included in this study were randomly assigned to 4 groups （50 rats each）： normal control group （NC）, uncontrolled diabetic group （DM1）, poorly-controlled diabetic group （DM2）, and well-controlled diabetic group （DM3）. Diabetic rats were induced by single intraperitoneal injection of streptozotocin, and the focal ischemic rat model of middle artery occlusion （MCAO） was made by insertion of fishing thread in 6 weeks after the establishment of the diabetic model. Each group was divided into 5 subgroups （10 rats each）： four focal ischemic subgroups at different ischemic-reperfusion time （at 3,12, 24 and 72 hours after reperfusion, respectively） and one sham-operated subgroup. The mRNA and protein expression of GLUT1 and GLUT3 was assessed by RT-PCR and Western blotting, respectively. Results There was significant difference in the mRNA expression of GLUT1 and GLUT3 between the four focal ischemic subgroups and the sham-operated subgroup at different reperfusion time in each group. The mRNA expression of GLUT1 and GLUT3 in the 4 ischemic groups began to increase at 3 hours, peaked at 24 hours after reperfusion and maintained at a higher level even at 72 hours compared with that of the sham-operated subgroup. The mRNA expression of GLUT1 increased more significantly than that of GLUT3. The mRNA expression of GLUT1 and GLUT3 was significantly different between the diabetic groups and normal control group. The mRNA expression of GLUT1 and GLUT3 was increased more significantty in the diabetic groups than that in the normal control group. There was a significant difference in the mRNA expression in the groups with different blood glucose levels. The mRNA expression tended to decrease with increased blood glucose levels. The expression trend of GLUT1 and GLUT3 protein was similar to that of GLUT1 and GLUT3 mRNA. Conclusions GLUT1 and GLUT3 expression was notably up-regulated in the penumbra region after cerebral ischemia in this study. But the up-regulated amplitude of GLUT1 and GLUT3 in the diabetic rats with cerebral ischemic injury became smaller than that of the normal controls. In the treatment of diabetic patients with cerebral embolism, blood glucose control should not be too strict, otherwise the up-regulation of GLUT1 and GLUT3 induced by cerebral ischemic injury might not be able to meet the needs of energy metabolism in cells. Chin Med J 2009; 122（ 17）： 1996-2001     

Relationship between AQP4 expression and structural damage to the blood-brain barrier at early stages of traumatic brain injury in rats

Background Although some studies have reported that aquaporin-4 （AQP4） plays an important role in the brain edema after traumatic brain injury （TBI）, little is known about the AQP4 expression in the early stage of TBI, or about the correlation between the structural damage to the blood-brain barrier （BBB） and angioedema. The aim of this project was to investigate the relationship between AQP4 expression and damage to the BBB at early stages of TBI. Methods One hundred and twenty healthy adult Wistar rats were randomly divided into two groups： sham operation group （SO） and TBI group. The TBI group was divided into five sub-groups according to the different time intervals： 1, 3, 6, 12, and 24 hours. The brains of the animals were taken out at different time points after TBI to measure brain water content. The cerebral edema and BBB changes in structure were examined with an optical microscopy （OM） and transmission electron microscopy （TEM）, and the IgG content and AQP4 protein expression in traumatic brain tissue were determined by means of immunohistochemistry and Western blotting. The data were analyzed with SPSS 13.0 statistical software. Results In the SO group, tissue was negative for IgG, and there were no abnormalities in brain water content or AQP4 expression. In the TBI group, brain water content significantly increased at 6 hours and peaked at 24 hours following injury. IgG expression significantly increased from 1 to 6 hours following injury, and remained at a high level at 24 hours. Pathological observation revealed BBB damage at 1 hour following injury. Angioedema appeared at 1 hour, was gradually aggravated, and became obvious at 6 hours. Intracellular edema occurred at 3 hours, with the presence of large glial cell bodies and mitochondrial swelling. These phenomena were aggravated with time and became obvious at 12 hours. In addition, microglial proliferation was visible at 24 hours. AQP4 protein expression were reduced at 1 hour, lowest at 6 hours, and began to increase at 12 hours, showing a V-shaped curve. Conclusions The angioedema characterized by BBB damage was the primary type of early traumatic brain edema. It was followed by mixed cerebral edema that consisted of angioedema and cellular edema and was aggravated with time. AQP4 expression was down-regulated during the angioedema attack, but AQP4 expression was upregulated during intracellular edema.     

Electro-acupuncture combined with transcranial magnetic stimulation improves learning and memory function of rats with cerebral infarction by inhibiting neuron cell apoptosis

This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism.Two hundred SD rats were randomly divided into four groups:normal group,sham-operated group,model group and EA+TMS group,and each group was then divided into five sub-groups in terms of the different time to start treatment post operation:6,12,24,48 and 72 h.Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R).After treatment for 14 d,the Morris water maze test was applied to examine the spatial learning and memory abilities of rats.In infarcted area,the expression of caspase-3 was immunohistochemically detected,and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA.The results showed that in EA+TMS group compared with model group at the same treatment time windows,the escape latency was substantially shortened,the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05).In the EA+TMS sub-groups,the escape latency was shortest,the expression level of caspase-3 lowest,and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h.It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3.The best time window is 24 h after perfusion treatment to ischemia.     

Protective effect of sulodexide on podocyte injury in adriamycin nephropathy rats

This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy （AN）. A total of 36 healthy male SD rats were randomly assigned to three groups： control group, AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin （6.5 mg/kg） in both AN group and sulodexide treatment group. Sulodexide （10 mg/kg） was administered the rats in the treatment group once daily by garage from the first day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14, 28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin, CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and protein were increased in the glomeruli of AN rats at day 14 and 28 after the model establishment, which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And, the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide-treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP （as well as 24-h urinary protein excretion）. It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.     

Effect of antimicrobial agents on the toll-like receptors and inflammatory cytokines in liver tissue of the alcohol-induced liver disease in rats with Vibrio vulnificus sepsis

Background Septicemia and inflammation-mediated septic shock caused by Vibrio vulnificus （VV） is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Methods Male Sprague-Dawley rats were assigned to the following treatment groups： normal control （N）, alcoholic liver disease control （A）, antimicrobial-treated alcoholic liver disease control （AA）, alcoholic liver disease with VV sepsis （AV）, and antimicrobial-treated alcoholic liver disease with VV sepsis （AVA）. Alcohol-induced liver disease was observed in all groups except N. Expression of mRNAs encoding hepatic toll-like receptors 2 and 4, myeloid differentiation protein-2, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and IL-10 was determined by RT-PCR. Results mRNAs encoding toll-like receptors 2 and 4 and myeloid differentiation protein-2 were significantly up-regulated in group AV as compared to control groups at 2-24 hours of sepsis; peak expression occurred at 12 hours. These mRNAs were also up-regulated in group AVA but to lesser degrees than in group AV at comparable time post-infection, mRNAs encoding TNF-α, IL-1β and IL-6 were significantly elevated in group AV as a function of infection. In group AVA as compared to AV, expression of TNF-α and IL-1β mRNAs was lower at 12-24 hours post-infection and expression of IL-6 mRNA was lower at 24 hours post-infection. Compared with control groups, IL-10 mRNA expression in group AV was markedly higher at 12-24 hours of sepsis. Expression of IL-10 mRNA was lower in group AVA as compared to AV at 24 hours of sepsis. Conclusions Antimicrobial therapy reduces expression of toll-like receptors and cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Monitoring hepatic toll-like receptor and cytokine expression during antibiotic therapy may be valuable for determining the course of VV sepsis in subjects with liver disease.     

针灸脑缺血再灌注损伤大鼠百会及足三里穴位对热休克蛋白70及肿瘤坏死因子-α表达的作用

Objective:To explore the effects of acupuncture on the peripheral serum expression of heat shock protein 70(HSP70)and tumor necrosis factor a(TNF-α)in rats with cerebral ischemia-reperfusion injury(CIRI).Methods:In total,152 Sprague-Dawley(SD)rats were randomly divided into an operated group and a non-operated group according to a random digits table.The operated group included a sham-operated group,a model group and an acupuncture group,whereas the non-operated group consisted of a normal group.Except for the normal group,each group was further divided into 12,24,48,72,96,and 144 h time points according to different reperfusion times.Eight rats were assigned in each operated group and in the normal group.The rat model of CIRI was established by the thread occlusion method in the model and acupuncture groups.The acupuncture group was treated with electroacupuncture at Baihui(DU20)and Zusanli(ST36)for the required time after successful operation.Blood was sampled to detect the HSP70 and TNF-αcontent by enzyme linked immunosorbent assay.Results:The expression of HSP70 protein in the peripheral serum of the experimental groups was higher than that in the normal control group.The peak time in both the model and the sham-operated groups was 12 h,and the peak time in the acupuncture group was 24 h.The expression in the acupuncture group declined to a lower level at 72 h and was lower than that in the model and sham-operated groups(P0.05).The peak time for the expression of TNF-αprotein in the peripheral serum of both the model and the acupuncture groups was 24 h,but the expression in the acupuncture group was lower than the model group.Additionally,the expression of TNF-αin all experimental groups was higher than the normal group(P0.05).Conclusions:Acupuncture at DU20 and ST36 in rats attenuated CIRI,which was associated with a reduction in the expression of HSP70 and TNF-α.These results provide clues to acupunctural neuroprotective properties.Acupuncture at DU20 and ST36 in rats after CIRI can adjust the expression of HSP70 and TNF-αin the peripheral serum,which might be one of the mechanisms of acupuncture's attenuation of CIRI.     

Contribution of Decreased Expression of Ku70 to Enhanced Radiosensitivity by Sodium Butyrate in Glioblastoma Cell Line（U251）

The present study investigated the enhanced radiosensitivity of U-251 cells induced by sodium butyrate(NaB) and its possible mechanisms.Increased radiosensitivity of U251 cells was examined by clonogenic cell survival assays.The expression of Ku70 mRNA and protein was detected by using RT-PCR and Western blotting respectively.γ-H2AX foci were measured at different time points after ionizing irradiation alone or combined with NaB treatment.The results showed that cell survival rate was significantly reduced,both D0 and Dq values were decreased(D0:1.43 Gy vs.1.76 Gy;Dq:1.22 Gy vs.2.05 Gy) after the combined treatment as compared with irradiation alone,and sensitivity enhancing ratio(SER) reached 1.23.The average number of γ-H2AX foci per cell receiving the combined treatment was significantly increased at different time points,and the expression levels of Ku70 mRNA and protein were suppressed by NaB in a dose-dependent manner.It was concluded that enhanced radiosensitivity induced by NaB involves an inhibited expression of Ku70 and an increase in γ-H2AX foci,which suggests decreased ability in DSB repair.     

Comparison of the Effects of Three Different Anti-fungus Drugs on Candida Albicans of Murine Vaginal Mucosa

To compare the therapeutic effects of three different anti-fungal drugs (i.e., terbinafine, fluconazole and intraconazole) in the treatment of experimental vaginitis caused by Candida albicans (C. albicans) in mice, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol. Mice were divided at random into different groups and then respectively treated with terbinafine, flucona- zole and intraconazole given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the begin- ning of the drug treatment. The fungal burdens in the vaginal lavage fluids taken at different time points from the mice treated with terbinafine were significantly higher than those taken at corre- sponding time points from mice treated with fluconazole or itraconazole (P<0.01). The fungal bur- dens in the vaginal lavage fluids taken from mice 1 week after the beginning of the treatment with terbinafine remained at a relatively high level. A dramatic drop in the fungal burden was noted in the vaginal lavage fluids taken on the 2nd day of the treatment from mice treated with itraconazole or flu- conazole group and the fungal burden on the 3rd day of the treatment in these mice were at a very low level, suggesting that fluconazole or itraconazole were highly effective for the treatment. How- ever, the difference in the therapeutic effect between the two drugs was not significant (P>0.05). Itraconazole or fluconazole, but not terbinafine, is very effective for the treatment of fungal vaginitis caused by C. albicans in mice.     

Effects of Acupuncture at Baihui(GV 20) and Zusanli(ST 36) on Peripheral Serum Expression of MicroRNA 124,Laminin and Integrin β1 in Rats with Cerebral Ischemia Reperfusion Injury

Objective:To explore the effects of acupuncture at Baihui(GV 20) and Zusanli(ST 36) on the peripheral serum expression of microRNA 124(miRNA 124),laminin and integrin β 1 in rats with cerebral ischemia reperfusion injury(CIRI).Methods:Seventy-two healthy male Sprague-Dawley rats were randomized into a model group,an acupuncture group,and a sham-operated group using a random digits table,with 24 rats per group.Each group was further randomly divided into 1-,3-,5-,and 7-day subgroups based on the reperfusion time according to a random digits table,with 6 rats in each subgroup.In the model and acupuncture groups,CIRI was induced using the thread occlusion method.Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations.Serum was sampled for detecting laminin and integrin β1 protein via enzyme-linked immunosorbent assay,and serum miRNA 124 was examined using quantitative polymerase chain reaction.Results:The serum level of miRNA 124 in the cerebral ischemia rats increased significantly,and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days.The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point(5.96 ±0.01 vs.3.11 ±0.04,P0.05).Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group.Compared with the model group,the level of laminin in the serum of the acupuncture group was significantly lower at each time point,especially at the 3-day,and 7-day time points(589.12±3.57 vs.793.05 ± 5.28,and 600.53 ±3.05 vs.899.06 ±5.74,P0.05).The level of integrin β 1 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and 7-day time points(208.66 ±0.95 vs.280.83 ±1.77,and212.36 ±0.95 vs.316.77±2.42,P0.05).Additionally,the model group and the acupuncture group showed dual peaks of integrin β 1 and laminin expression at 3-day and 7-day.Conclusions:Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin β 1 and laminin in peripheral serum.These changes may represent one of the mechanisms underlying acupuncture's attenuation of CIRI.     

PARP-1激活和AIF易位在肠缺血再灌注损伤中的作用

目的:研究大鼠肠缺血再灌注后肠损伤中是否存在聚腺苷二磷酸核糖聚合酶-1(PARP-1)激活和凋亡诱导因子(AIF)易位,并探索二者在肠缺血再灌注损伤中的作用?方法:通过阻断腹腔干和肠系膜上动脉30 min后,恢复血运,制作鼠肠缺血再灌注模型,分为肠缺血再灌注组(I/R),缺血再灌注前15min经静脉给PARP-1抑制剂3-AB组(Drug)和假手术组(Sham)?取肠组织分别作HE染色?聚腺苷二磷酸核糖(PAR)的免疫组化染色,TUNEL方法检测组织细胞凋亡情况,Western Blot检测凋亡早期信号PARP-1片段p85和凋亡诱导因子(AIF)的表达情况?结果:I/R组肠损伤程度?PAR阳性表达率?细胞凋亡率和凋亡早期信号PARP-1 p85片段?AIF表达程度均较Sham组显著增高(P < 0.05);应用PARP-1抑制剂后,Drug组上述指标均较I/R组显著降低(P < 0.05),但仍高于Sham组?结论:在大鼠肠缺血再灌注损伤中存在PARP-1激活和AIF易位两分子事件,并在肠缺血再灌注损伤中发挥重要作用;肠缺血再灌注前应用PARP-1抑制剂有肠保护效果?     

3-AB对Hela细胞PARP表达及X线照射后Hela细胞凋亡和周期进程的影响

目的：研究多聚（ADP-核糖）聚合酶(PARP)特异性抑制剂3-氨基苯甲酰胺（3-AB）抑制PAR后Hela细胞凋亡和周期进程的变化及PARP对电离辐射损伤细胞的作用机制。方法：采用体外传代培养的人宫颈癌上皮细胞Hela细胞为实验材料；给予Hela细胞3-AB( 5 mmol• L¹) 0、2、4、8及12 h后，采用PARP单克隆抗体荧光标记，流式细胞术检测对照组及3-AB组经3-AB作用不同时间后Hela细胞中PARP蛋白表达；在给予3-AB 2 h后，对阴性对照组、单纯X线照射组及3-AB加X线照射组Hela细胞行2 Gy X线照射2、8、12及24 h后应用流式细胞术测定 Hela细胞凋亡和细胞周期进程。结果：给予3-AB后2、4、8及12 h PARP蛋白表达阳性的Hela细胞百分数均明显低于阴性对照组（P<0.01），其中在2 h时降低较明显，阳性细胞率为(7.60±1.36)%。进行2 Gy照射后2、8、12及24 h各时间点，3-AB加X线照射组Hela细胞凋亡率均明显高于单纯X线照射组（P<0.01或P<0.05），而G₂期细胞百分数均低于单纯X线照射组（P<0.01或P<0.05）。结论：3-AB明显抑制Hela细胞PARP表达，增加电离辐射所致细胞凋亡并减少G₂期细胞阻 滞。     

3-氨基苯甲酰胺影响糖尿病大鼠晶状体上皮细胞NF-κB表达的免疫组化研究

目的观察聚腺苷二磷酸核糖聚合酶(PARP)抑制剂3-氨基苯甲酰胺(3-AB)对早期糖尿病(DM)大鼠模型晶状体上皮细胞中核因子κB(NF-κB)表达的影响。方法采用一次性腹腔注射1%链脲佐菌素(STZ)50 mg/kg建立DM大鼠模型。成模后将大鼠随机分为正常对照组(N组)、糖尿病组(DM组)和干预组(DM+P组)。自建立模型后第3天起,DM+P组每日给予3-AB 30 mg/kg灌胃,N组和DM组每天给予等体积0.9%生理盐水灌胃。分别于给药后2、4、8周处死各组大鼠,取出晶状体,免疫组化SP法检测晶状体上皮细胞中NF-κB P65的表达情况。结果 DM 2、4、8周组大鼠晶状体上皮细胞NF-κB P65表达较相应时间点的N组高;DM+P 2、4、8周组大鼠晶状体上皮细胞NF-κB P65表达较相应时间点的N组高;DM+P 2、4、8周组与相应时间点DM组比较,晶状体上皮细胞NF-κB P65表达降低。结论 PARP抑制剂3-AB可以抑制早期糖尿病大鼠晶状体上皮细胞中NF-κB的表达。     

急性心肌梗死猝死者心肌细胞hsf1和HSP70改变的意义

目的探讨急性心肌梗死猝死者梗死区心肌细胞内热休克转录因子1(hsf1)和热休克蛋白70(HSP70)改变的临床意义。方法分别用RT-PCR和免疫组化法检测(IHC)18例急性心肌梗死猝死者(研究组)和15例心脏正常因车祸快速死亡者(对照组)心肌细胞中hsf1和HSP70基因的mRNA和蛋白表达量。结果急性心肌梗死猝死者心肌细胞hsf1和HSP70的mRNA表达量都显著高于正常对照组(P<0.01),且hsf1和HSP70 mRNA的表达量之间呈显著的正相关关系(P<0.001)。急性心肌梗死猝死者心肌细胞hsf1和HSP70蛋白在细胞浆和细胞核表达较对照组显著增强(P<0.01),其中hsf1蛋白主要在心肌细胞核内表达,HSP70蛋白主要在心肌细胞浆内表达。结论急性心肌梗死猝死者心肌细胞内hsf1和HSP70可能共同参与了急性心肌梗死的病理生理过程,这一过程可能是热休克反应的另一调节途径。     

β受体阻滞剂阿替洛尔和酒石酸美托洛尔对大鼠急性心肌梗死后心肌细胞凋亡及凋亡相关基因表达的作用

目的比较阿替洛尔和酒石酸美托洛尔对大鼠急性心肌梗死(AMI)后心肌细胞凋亡及凋亡相关基因表达的作用。方法251只雌性SD大鼠结扎左冠状动脉建立AMI模型,术后24h存活的124只随机分为AMI对照(MI组,n=43)、阿替洛尔(A组,10mg·kg-1·d-1,n=39)和酒石酸美托洛尔(B组,20mg·kg-1·d-1,n=42)治疗组;另设假手术组(S组,n=27)。各组再按观察时点随机分为48h和4周两亚组。术后24h以直接灌胃法给药。末端脱氧核苷酸转移酶介导的dUTP切口末端标记技术(TUNEL)和DNA凝胶电泳检测心肌细胞凋亡。免疫组织化学方法和Western blot检测“凋亡抑制基因”bcl-2、“凋亡促进基因”bax和“凋亡执行因子”caspase-3基因的表达。结果与AMI对照组相比,AMI后48h,A、B两组梗死区、边缘区和非梗死区的心肌细胞凋亡指数,除B组梗死区显著降低(P<0·01)外,其他指标差异均无显著性(均P>0·05);心肌细胞中bcl-2的表达除A组的非梗死区外均增加(免疫组织化学染色),bax和caspase-3的表达均无明显降低。AMI4周时,A、B两组瘢痕区及其边缘区和非梗死区的心肌细胞凋亡指数均显著降低(P<0·05,P<0·01);bcl-2、bax和caspase-3的表达均无明显变化,仅A4周组非梗死区bax的表达明显降低。Western blot显示,与AMI对照组相比,A、B两组心肌细胞中caspase-3、bcl-2和bax的表达差异均无显著性,但bcl-2/bax的比值显著增加(P<0·05),并与假手术组相当。结论阿替洛尔和酒石酸美托洛尔均能减少AMI梗死/瘢痕区、边缘区和非梗死区的心肌细胞凋亡,作用相当,此作用主要是通过增加bcl-2的表达和bcl-2/bax的比值而实现。     

抑郁对心肌梗死大鼠缺血心肌细胞凋亡及Bcl-2、Bax和Caspase-3表达的影响

目的:探讨抑郁对心肌梗死大鼠缺血心肌细胞凋亡及Bcl-2、Bax和Caspase-3的影响。方法:大鼠随机分为四组,各组8只,A组为假手术组,B组为抑郁大鼠假手术组,C组为心肌梗死组,D组为抑郁大鼠心肌梗死组。用TUNEL法检测心肌细胞凋亡,用免疫组织化学法和逆转录-聚合酶链反应方法检测Bcl-2、Bax和Caspase-3的基因表达。结果:各组大鼠心肌细胞凋亡指数以及心肌细胞Bcl-2、Bax、Caspase-3蛋白表达和心肌细胞Bcl-2、Bax、Caspase-3 mRNA表达的差异有统计学意义(P<0.01),C、D组大鼠心肌细胞凋亡指数以及心肌细胞Bcl-2、Bax、Caspase-3蛋白表达和心肌细胞Bcl-2、Bax、Caspase-3 mRNA表达均高于A组和B组,差异有统计学意义(P<0.01),D组大鼠心肌细胞凋亡指数以及心肌细胞Bax、Caspase-3蛋白表达和心肌细胞Bax、Caspase-3 mRNA表达高于C组,而心肌细胞Bcl-2蛋白及mRNA表达低于C组,差异有统计学意义(P<0.05)。结论:抑郁可以加重心肌梗死大鼠缺血心肌细胞凋亡,其机制可能与上调促凋亡基因Bax及Caspase3表达、下调抗凋亡基因Bcl-2的表达有关。     

miR-126对急性心肌梗死大鼠心肌细胞凋亡的影响

目的探讨miR-126对急性心肌梗死(AMI)大鼠心肌细胞凋亡的影响。方法通过冠状动脉左前降支结扎大鼠建立AMI模型,随机分为AMI组、miR-126 mimics negative control(NC)组及miR-126组,其中NC组及miR-126组分别采用心肌组织局部注射慢病毒转染miR-126 mimics NC及miR-126 mimics,另外假手术组采用冠状动脉左前降支穿线不结扎。记录大鼠心脏功能,TCC法及原位末端凋亡法(TUNEL)检测心肌凋亡指数及梗死面积,比色法检测天冬氨酸蛋白水解酶(Caspase 3)及Caspase 8活性,Western-blot法检测Bax及Bcl-2表达,同时RTPCR法检测Fas及Fas-L mRNA表达。结果与假手术组比较,AMI组及NC组左室射血分数(LVEF)及左室长轴缩短分数(FS)升高,左室舒张末期内径(LVDd)及左室收缩末期内径(LVDs)降低,心肌凋亡指数提高,心肌梗死面积增大,Caspase 3及Caspase 8活性上升,Bax蛋白表达量上调,Bcl-2蛋白表达量下调,Fas及Fas-L mRNA水平上升,差异具有统计学意义(P0.01)。与AMI组及NC组比较,miR-126组能扭转此变化,差异具有统计学意义(P0.01)。结论 miR-126能显著抑制AMI大鼠心肌细胞凋亡,与调节细胞凋亡相关蛋白表达有关。     

中介素抑制大鼠心肌梗死后心室重构

目的利用外源性中介素(IMD)治疗急性心肌梗死(AMI)后大鼠,探讨其是否具有抑制心室重构的作用及可能的机制。方法结扎大鼠左冠状动脉前降支制备心肌梗死模型。观察模型大鼠存活情况,超声心动图检测左室舒张期末内径(LVEDD)和左室收缩期末内径(LVESD),Western blot检测胞核内核因子-κB(NF-κB)及胞浆内核因子-κB抑制蛋白α(IκBα)的蛋白表达。结果与假手术组比较,AMI组LVEDD、LVESD均显著升高,与AMI组比较,IMD治疗组LVEDD、LVESD均显著降低(P<0.01或0.05)。与假手术组比较,AMI组NF-κBp65蛋白明显增加,而IMD治疗组NF-κBp65蛋白表达较AMI组明显降低。与假手术组比较,AMI组IκBα蛋白表达明显降低,而IMD治疗组IκBα蛋白表达较AMI组明显增加。结论 IMD能够明显减少AMI后大鼠心肌NF-κB表达,减轻梗死后左室扩张程度。     

大鼠视网膜缺血再灌注后多聚二磷酸腺苷核糖聚合酶的表达变化

目的观察大鼠视网膜缺血再灌注损伤后不同时相视网膜结构变化及多聚二磷酸腺苷核糖聚合酶(PARP)的表达情况。方法升高眼压到110mmHg持续1h，建立大鼠视网膜缺血再灌注模型。HE染色观察视网膜组织病理学改变；免疫组化Ehvision^TM法检测视网膜细胞PARP表达。结果正常对照组大鼠视网膜仅见微量PARP表达；缺血1h再灌0h组视网膜结构未见明显变化；再灌注6h，视网膜开始有中性粒细胞浸润，神经节细胞层(ganglion cell layer，GCL)PARP表达显著增加；12、24h组视网膜中有较多中性粒细胞浸润，神经节细胞层PARP表达24h达高峰后下降，但仍维持较高水平；24h后内核层(inner nuclear layer，INL)细胞PARP表达明显增高，168h达高峰，伴之神经节细胞层和内核层细胞明显变薄。另外，也见自身对照眼视网膜神经节细胞层PARP表达增加。结论大鼠视网膜缺血再灌注早期PARP即表达上调，24h组在GCL达高峰，168h组在INL达高峰，这一过程可能与视网膜神经节细胞层和内核层细胞凋亡有关。     

氯化镉对大鼠心肌H9c2细胞DNA损伤的作用机制

目的：研究氯化镉（CdCl2）在体外对大鼠心肌细胞DNA的损伤作用,探讨CdCl2心肌毒性作用机制。方法：以0、5、10、30、50和80 μmol?L-1 CdCl2作用于大鼠心肌H9c2细胞,采用单细胞凝胶电泳法（SCGE）检测作用6、12及24 h的DNA损伤,Western blotting法检测CdCl2作用24 h对聚ADP-核糖聚合酶（PARP）的影响,免疫细胞化学法检测细胞色素C（Cyt-C）和凋亡诱导因子（AIF）蛋白表达变化的情况。结果：随CdCl2 浓度的增大和作用
时间延长,作用12、24 h后,10、30、50和80  μmol/L剂量组大鼠心肌细胞DNA损伤程度明显,组间比较差异有统计学意义（P< 0.05或P< 0.001）。作用24 h后,30、50和80  μmol?L-1剂量组可见明显PARP裂解89 000片段。Cyt-C和AIF蛋白的表达也随着CdCl2浓度的加大而显著增高,组间比较差异有统计学意义（P< 0.05或P< 0.001）。结论：CdCl2对大鼠心肌细胞的毒性作用机制可能与诱导DNA损伤,影响PARP、Cyt-C和AIF蛋白表达有关,且具有一定的剂量依赖关系。