Human neural apoptosis and regulation mechanism of apoptosis-related genes following traumatic brain injury

Objective To observe human neural apoptosis and undestand regulation mechanism and expression changes of apoptosis-related genes posterior to traumatic brain injury (TBI) so as to explore a fresh therapy for delayed neural death. Methods The specimens from patients with acute TBI were collected to observe the DNA injury by means of light,electrical micropoopes and TUNEL after TBI. By RT-PCR, in situ hybridization and immunohistochemistry, the expression changes of bcl-2, bax, p53 and caspase-3 p20 subunit at mRNA and protein level were observed. Results Apoptotic neurons occurred following TBI and peaked atChina Medical Abstracts(Surgery) the 24th hour. The normal brain tissues rarely contained mRNA or protein of bcl-2 and p53 or activated caspase-3. In a short time after TBI, the mRNA and protein expressions of bcl-2 and p53 increased and caspase-3 was activated. The primary expressions of bax mRNA and bax protein were continuously high in the normal brain tissues, After TBI, the expression level of ba     

Role of PI3K/Akt signaling in the protective effect of magnesium sulfate against ischemia-perfusion injury of small intestine in rats

Background The protective effects of magnesium sulfate against ischemia-reperfusion injury of the small intestine in Sprague-Dawley （SD） rats have been confirmed in our previous research. However, its exact mechanism is unclear. This study was to evaluate the role of PI3K/Akt signal pathway in the protective effect of magnesium sulfate against ischemia-reperfusion injury of the small intestine in SD rats. Methods Rat model of intestinal ischemia-reperfusion injury was used. The SD rats were divided into four groups randomly： sham operation group, ischemia-reperfusion group, magnesium sulfate group and magnesium sulfate plus LY294002 （an inhibitor of PI3K） group. The pathological changes of intestinal mucosa were examined; the activity of diamine oxidase （DAO） in plasma, the plasma contents of malondialdehyde （MDA）, and apoptosis rate of the intestinal mucosal cells were determined and compared. The expression of p-Akt was detected by Western blotting. Results There were more evident pathological changes of the intestinal mucosa （higher Chiu＇s score, P 〈0.05）, enhanced DAO activity （P 〈0.05）, elevated contents of MDA （P 〈0.05）, higher apoptosis rate （P 〈0.05）, and lower level of p-Akt （P 〈0.05） in the ischemia-reperfusion group compared with the sham operation group. There were less evident pathological changes of the intestinal mucosa （lower Chiu＇s score, P 〈0.05）, lower DAO activity （P 〈0.05）, lower contents of MDA （P 〈0.05）, and lower apoptosis rate （P 〈0.05）, but higher level of p-Akt （P 〈0.05） irL the magnesium sulfate group compared with the ischemia-reperfusion group. There were more evident pathological changes of the intes,inal mucosa （higher Chiu＇s score, P 〈0.05）, higher contents of MDA （P 〈0.05）, higher DAO activity （P 〈0.05） and higher apoptosis rate （P 〈0.05）, and lower level of p-Akt （P 〈0.05） in the magnesium sulfate plus LY294002 group compared with the magnesium sulfate group. Conclusions Activation of PI3K/Akt signal pathway results in the reduction of cell apoptosis, which likely accounts for the protective effect of magnesium sulfate against intestinal ischemia-reperfusion injury.     

Effects of Buyang Huanwu Decoction on Apoptosis of Nervous Cells and Expressions of Bcl-2 and Bax in the Spinal Cord of Ischemia-Reperfusion Injury in Rabbits

To study the effect of Buyang Huanwu Decoction （补阳还五汤BYHWD） on spinal ischemia-reperfusion injury and explore the possible mechanism involved. Method： 27 model rabbits with ischemia-reperfusion injury were randomly divided into a sham operation group, a model group and a BYHWD-pre-treated group. 24 and 48 hours after the ischemia-reperfusion injury, the motor functions of hind limbs were observed, the expressions of apoptosis-related proteins bcl-2 and Bax in the spinal tissue were investigated with the immunohistochemical methods and apoptosis of nervous cells with TUNEL straining; and the malondialdehyde （MDA） level and superoxide dismutase （SOD） activity in the spinal cord were then determined. Results： BYHWD can improve significantly the motor function of hind limbs, increase SOD activity and decrease the level of MDA, up-regulate expression of Bcl-2 and down-regulated expression of Bax, and depress apoptosis of nervous ceils in the spinal cord caused by the ischemia reperfusion in the rabbits. Conclusion： BYHWD has a protective action on the spinal ischemia-reperfusion injury, and the mechanisms may be related to the decrease of lipid peroxidation and inhibition of apoptosis of nervous cells.     

Effects of Carvedilol on Cardiomyocyte Apoptosis and Gene Expression In Vivo after Ischemia-Reperfusion in Rats

The effects of carvedilol on cardiomyocyte apoptosis and expression of bcl-2, bax genesfollowing ischemia (0.5 h) and reperfusion (48 h) in vivo and the possible biological mechanism ofcarvedilol inhibiting cardiomyocyte apoptosis were studied. The left anterior descending artery inWistar rats were ligated to establish ischemia-reperfusion (I/R) models. The model animals were di-vided into two groups: I/R group, the model rats not subject to other treatments except ischemia-reperfusion (n=8); carvedilol-treated group (n=8), I/R model rats treated with carvedilol. Eightrats in the sham-operated group were subjected to only experimental open operation. The number ofapoptotic cardiomyocyte was determined by TUNEL staining. Immunohistochemistry and in situ hy-bridization histochemistry (ISHH) were used to detect the expression of bcl-2 and bax genes. Imageprocessing system was used to quantitatively dispose the positive metric substances of both immuno-histochemistry and ISHH through the average optical den     

Effect of Shenfu on the Expression of Bcl-2 and Bax of Intestinal Mucosal Epithelial Cells after Ischemic Reperfusion in Rats

Objective To study the effect of Shenfu on the expression of bax and bcl-2 in small intestinal mucosal epithelial cells after ischemic reperfusion injury and explore the effect of Shenfu on small intestinal mucosal epithelial cells apoptosis. Methods 36 SD rats were randomized into three groups, each consisting of 12 rats： Sham group （S group）, Ischemic reperfusion group （IR group）, Shenfu group （SF group）, Ischemic reperfusion models were made by ligated the superior menseneric artery for 1 hour followed by 2 hrs reperfusion. Histological mucosal damage in each group was graded according to Chiu＇s score. Immunohistochemistry detected the expression of bax and bcl-2, and obtained the optical density （OD） value using a color image pattern analysis system, and then calculated the ratio of bcl-2/bax. TUNEL method measured apoptotic intestinal mucosal epithelial cells, and calculated the apoptotic index. Results There were edema and epithelial impairment in part of villus and the integral was higher in SF group than S group （P〈0.05）, but lower than IR group （P〈0.01）. Compared IR group with S group and SF group, the average OD value of bcl-2 and bax had significant statistic difference （P〈0.01）, and the average OD value of bcl-2 in SF group was higher than in S group （P〈0.05）, meanwhile bcl-2/bax was significantly lower in IR group than in S group and SF group （P〈0.01）, but that was higher in SF group than in S group （P〈0.05）. Apoptotic index in IR group was significantly higher than that in other two groups （P〈0.01）, and that in SF group was higher than in S group （P〈0.05）. Conclusion Shenfu can enhance the expression of bcl-2, decrease the expression of bax, meanwhile increase the ratio of bcl-2/bax, inhibit small intestinal mucosal epithetlial cells apoptosis, and protect small intestine mucosal epithetlial after ischemic reperfusion.     

Focal cerebral ischemia induces Alzheimer’s disease-like pathological change in rats

The changes in the tau protein phosphorylation and expression of bcl-2,and bax in rat parietal cortex neurons after focal cerebral ischemia-reperfusion(I/R)were explored,and the relationship between the tau protein phosphorylation and the expression of bax or apoptosis was clarified in order to elucidate the relationship between cerebral infarction and Alzheimer's disease.The rat focal cerebral I/R model was induced by occlusion of the right middle cerebral artery using the intraluminal suture method.The le...     

Expression of NF-kB in Schwann apoptosis in spinal cord following cells and its effect on motor neuron sciatic nerves injury in rats

Objective：To explore the expression of nuclear factor-kappa B （NF-kB） in Schwann cells （SCs） and its effect on motor neuron apoptosis in spinal cord following sciatic nerves injury in adult rats. Methods： Thirty-six adult Sprague-Dawley （SD） rats were divided randomly into normal control group （n=6）, and sciatic nerves crushing group （n= 30）, and the later was further equally randomized into 5 subgroups： 1, 3, 7, 14, and 21 d post-injury groups. The expression of NF-kB of normal and injured nerves were examined by immunohistochemistry staining, and the apoptosis of motor neurons in spinal cord of lumbar 4 to lumbar 6 （L4-L6） was investigated by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling （TUNEL） assay. Both were qua.ntitated by image analysis. Results： In crushing group, except 21 d post-injury group, the expression of NF-kB was markedly higher than that in the normal control group （P〈0.05, P〈0. 01）. At 1 d after sciatic nerves crushing, the expression of NF-kB was obviously up-regulated, reached peak at 3 d, and recovered at 21 d. The same trend was observed in the time-course on motor neuron apoptosis after sciatic nerves injury. Correlation analyses revealed that motor neuron apoptosis was significantly and positively correlated with the expression of NF-kB following sciatic nerves injury （r= 0. 976 0, P〈0. 01）. Conclusion： After injury of sciatic nerves, the presence and up-regulation of NF-kB in SCs may be involved in motor neuron apoptosis in L4-L6 spinal cord.     

Evidence of apoptotic smooth muscle cells in proliferative intima of injured arteries

Objective To investigate the occurrence and extent of apoptosis in the course of restenosis. Methods The experimental models of vessel narrowness and intima thickness were established in minipigs’ iliac arteries by balloon injury and specimens were retrieved on the 1st, 3rd, 6th,12th and 30th days for dynamic observation. Apoptotic smooth muscle cells (SMCs) were detected by terminal deoxynucleotidyl transferase- mediated dUTP nick- end labeling (TUNEL). Results Apoptotic SMCs occurred only in the thickened intima 12 days after injury accompanied with the proliferative SMCs, the percentage of apoptosis was 1. 94%±0. 42% on the 12th day and 1. 36%±0. 31% on the 30th day respectively. The low frequency of apoptosis compared with the proliferative SMCs was a feature in the restenotic pathology. Conclusions Apoptosis participates in the pathogenetic process of intimal thickening and its level was low compared with proliferation. The findings suggest that attempts to modulate apoptosis after vessel injury constitute a theoretical approach to the prevention of restenosis.     

Expression of NF-κB in Schwann cells and its effect on motor neuron apoptosis in spinal cord following sciatic nerves injury in rats

Objective: To explore the expression of nuclear factor-kappa B (NF-κB) in Schwann cells (SCs) and its effect on motor neuron apoptosis in spinal cord following sciatic nerves injury in adult rats. Methods: Thirty-six adult Sprague-Dawley (SD) rats were divided randomly into normal control group (n=6), and sciatic nerves crushing group (n=30). and the later was further equally randomized into 5 subgroups: 1. 3. 7. 11. and 21 d post-injury groups. The expression of NF-κB of normal and injured nerves were examined by immunohistochemistry staining, and the apoptosis of motor neurons in spinal cord of lumbar 4 to lumbar 6 (L4-L6) was investigated by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNED assay. Both were quantitated by image analysis. Results: In crushing group, except 21 d post-injury group, the expression of NF-κB was markedly higher than that in the normal control group (P<0. 05,P<0. 01). At 1 d after sciatic nerves crushing, the expression of NF-κB was obviously up-regulated, reached peak at 3 d. and recovered at 21 d. The same trend was observed in the time-course on motor neuron apoptosis after sciatic nerves injury. Correlation analyses revealed that motor neuron apoptosis was significantly and positively correlated with the expression of NF-κB following sciatic nerves injury (r=0. 976 0,P<0. 01). Conclusion: After injury of sciatic nerves, the presence and up-regulation of NF-κB in SCs may be involved in motor neuron apoptosis in L4-L6 spinal cord.     

Effects of rHu-EPO on Myocyte Apoptosis and Cardiac Function Following Acute Myocardial Infarction in Rats

The mechanisms of rHu-EPO attenuating the apoptosis after myocardial infarction in rats were studied. Thirty-two rats were divided into three groups: sham operation group (Sham), acute myocardial infarction group (MI) and rHu-EPO-treated group (MI EPO). Acute myocardial infarction model was made by ligating the anterior descending coronary artery, rHu EPO was administered i.p. in MI EPO group at the dose of 5 000 IU/kg body weight immediately after the ligation. Each rat in MI EPO group received the same dose of rHu-EPO daily the next 6 days. On the 14th day all rats underwent hemodynamic measurements and then killed. The samples were examined with HE stain, immunohistochemistry technique (bcl-2, bax) and TUNEL dyeing. The results showed that hemodynamic function in MI.4- EPO group was much better than in MI group.The number of the cells positive for bax and TUNEL in MI EPO group was less than that in MI group. The number of the cells positive for bcl-2 in MI EPO group was more than that in MIgroup. These findings suggested that rHu-EPO could treat myocardial infarction by preventing apoptosis and attenuating post-infarction deterioration in hemodynamic function.     

Effects of L-Tetrahydropalmatine on the Expressions of bcl-2 and bax in Rat after Acute Global Cerebral Ischemia and Reperfusion

Polygenesregulateapoptosis .bcl 2isananti apoptosisgene ,whilebaxisapro apoptosisgene .L Tetrahydropalmatine (L THP) ,akindofalkaloidextractedfromtraditionalChinesemedicineRhizomacorydalis,possessesanalgesic ,sedativeandhypnoticeffects.Recently ,ithasbeen…     

经颈内动脉注射pLXSN-bcl-2对脑局灶性缺血再灌注大鼠脑组织的影响

目的:研究经颈内动脉注射pLXSN-bcl-2对大鼠脑缺血再灌注损伤后的影响。方法:51只健康成年Wistar大鼠随机分对照组、空质粒组、bcl-2组,每组分缺血再灌注9,24h两小组。采用线栓法制作大脑中动脉梗死模型,2h后实现再灌注。3h后经颈内动脉注射质粒pLXSN,pLXSN-bcl-2。检测缺血再灌注24h后大鼠脑梗死体积,及各组大鼠脑内bcl-2、caspase-3的表达情况和神经细胞凋亡情况。结果:bcl-2组缺血再灌注24h的脑梗死体积最小;bcl-2组在缺血再灌注9,24h后bcl-2阳性表达明显升高;缺血再灌注24h caspase-3蛋白表达及凋亡细胞数明显降低,缺血再灌注9h后与其他组之间无差异。结论:脑缺血再灌注3h后经颈内动脉注射pLXSN-bcl-2可以增加bcl-2在脑内的表达,并在缺血再灌注24h后通过抑制caspase-3的表达,抑制神经细胞凋亡,减少脑梗死体积。而在缺血再灌注9h尚未表现出明显的神经保护作用。     

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。     

糖尿病结肠动力障碍大鼠远端结肠平滑肌细胞bax、bcl-2及caspase-3的表达

目的:探讨糖尿病(DM)结肠动力障碍大鼠远端结肠平滑肌细胞凋亡基因的表达。方法:雄性SD大鼠18只应用链脲佐菌素40mg/kg尾静脉注射后成功建立DM结肠动力障碍大鼠模型,分为模型6周和10周组;另18只分为正常对照6周和10周组,各组9只大鼠。应用实时荧光定量PCR检测各组大鼠远端结肠平滑肌细胞的bax、bcl-2和caspase-3mRNA表达,采用免疫组化SP法检测其蛋白的表达。结果:不同组别和作用时间各组大鼠远端结肠平滑肌细胞bax、bcl-2、caspase-3mRNA和蛋白的表达差异均有统计学意义(P均<0.001),模型组与同一时间点正常对照组以及模型10周组与6周组大鼠比较,远端结肠平滑肌细胞bax和caspase-3mRNA和蛋白的表达增强,bcl-2mRNA和蛋白的表达降低(P<0.05)。结论:远端结肠平滑肌细胞凋亡基因bax、bcl-2及caspase-3表达的改变可能是DM大鼠结肠动力障碍发病机制之一。     

雌二醇对大鼠脑缺血/再灌注损伤脑组织bcl-2、caspase-3表达的影响

目的：观察雌二醇时大鼠脑缺血／再灌注损伤脑组织hcl-2．capase-3表达的影响。方法：Wistar大鼠72只，随机分为假手术组、局灶性脑缺血2h／再灌注3h、6h．12h、24h组及雌二醇治疗(100μg／kg) 脑缺血2h／再灌注3h、6h、12h、24h组，每时间点8只，假手术组8只。采用线栓法建立大鼠局灶性脑缺血／再灌注损伤模型，采用连续切片免疫组化技术检测脑组织中bel-2、caspase-3表达部位及数量。结果：雌二醇治疗组皮层bcl-2免疫阳性细胞数与脑缺血／再灌注及似手术组比较，在再灌注后12h、24h增高，而纹状体bcl-2免疫阳性细胞数与脑缺血／再灌注组及假手术组比较筹异无统计学意义。假手术组无caspase-3表达，雌二醇治疗组caspase-3阳性细胞数，在皮层12h、24h明显减少．纹状体区caspase-3阳性细胞数治疗组各时间点稍减少，但与脑缺血／再灌注组比较无统计学意义。结论：雌二醇治疗可以诱导皮层bcl-2表达及下调caspase-3活性。     

盐酸戊乙奎醚预处理对大鼠局灶脑缺血再灌注损伤及凋亡相关基因caspase-3的影响

目的:观察盐酸戊乙奎醚预处理对大鼠局灶脑缺血再灌注损伤的影响及作用机制。方法:将60只雄性SD大鼠随机分为 3 组(每组20只)。对照组（C组）：进行假手术操作,未使用线栓法建立局灶脑缺血模型;缺血-再灌注组（I-R组）：麻醉前30 min腹腔内注射生理盐水2.0 mg•kg^-1后,使用线栓法建立大鼠局灶脑缺血模型,缺血120 min后进行再灌注;盐酸戊乙奎醚预处理组（P组）：麻醉前30 min腹腔内注射盐酸戊乙奎醚2.0 mg•kg^-1后,用线栓法建立大鼠局灶脑缺血模型,缺血120 min后进行再灌注。氯化三苯基四氮唑(TTC)染色观察测定脑梗塞体积;HE 染色法观察大鼠海马神经细胞形态的变化;半定量RT-PCR法测定海马组织caspase-3 mRNA的表达水平。结果:与I-R组比较,P组脑梗塞体积显著减小（P< 0.05）;I-R组海马神经细胞发生严重损伤,细胞核固缩、浓染,P组海马神经细胞损伤显著减轻;I-R组和P组caspase-3 mRNA表达水平明显高于C组（P< 0.05）,但P组caspase-3 mRNA表达水平低于I-R组（P< 0.05）。结论：盐酸戊乙奎醚预处理可以显著改善大鼠局灶脑缺血再灌注后海马神经细胞损伤,从而对脑缺血再灌注损伤发挥保护作用,这种保护作用可能与盐酸戊乙奎醚抑制大鼠脑海马组织凋亡蛋白酶caspase-3 mRNA的表达有关。     

Neuregulin-1β对阿霉素致心衰大鼠心肌细胞凋亡及bax、bcl-2蛋白表达的干预效应

目的 观察阿霉素所致心衰大鼠心肌细胞凋亡和bax、bcl-2蛋白表达情况及Neuregulin-1β(NRG-1β)的干预作用.方法 60只雄性SD大鼠,体质量200-250 g,分成3组:①模型组,即阿霉素心肌病组(ADR-DCM,n=20),阿霉素2 mg/kg,尾静脉注射,每周1次,连续注射8周;②阿霉素心肌病+NRG-1治疗组(NRG,n=20),在注射ADR后8周后即丌始给予NRG-1β 10μg(kg·d)尾静脉注射,连续5 d;③正常对照组(n=20),用2 ml/kg生理盐水,尾静脉注射,连续8周.9周后取心肌行HE染色病理观察,TUNEL法观察心肌细胞凋亡,Western blot检测bax、bcl-2蛋白的表达.结果 模型组与NRG治疗组心肌细胞凋亡指数较正常对照组显著增高(P<0.05),与模型组相比,NRG治疗组凋亡指数显著降低(P<0.05).与模型组比较,NRG治疗组bcl-2蛋白表达明显上调(P<0.05),bax蛋白表达明显下调(P<0.05).结论 NRG-1β能够抑制阿霉索所致心衰大鼠心肌细胞凋亡,其机制可能与bcl-2蛋白表达上调、bax蛋白表达下调、bcl-2/bax值上调有关.     

电针预处理对大鼠肝脏缺血再灌注损伤中肝细胞凋亡的影响

目的电针预处理,对大鼠肝脏缺血再灌注损伤中肝细胞凋亡的影响。方法24只健康雄性Wistar大鼠,随机分为3组,即假手术组(C)、电针刺预处理+缺血再灌注组(EAP+IR)、单纯缺血再灌注组(IR)。EAP+IR、IR组采用Pringle法建立肝缺血再灌注损伤模型,EAP+IR组于术前电针刺激阳陵泉穴30 min。再灌注6 h后,取部分肝组织用于原位DNA末端标记法(TUNEL)检测肝细胞凋亡率的变化;免疫组化测肝组织中bax和caspase-3的表达;WESTERN-BLOT法对肝脏中bcl-2、bax和caspase-3的表达。结果与C组比较,EAP+IR、IR组各种蛋白表达均增加,肝细胞凋亡率也明显升高。与IR组相比,EAP+IR组中肝细胞的凋亡率明显下降(P<0.05),caspase-3和bax蛋白表达量降低,而bcl-2蛋白则明显上调,免疫组化与WESTERN-BLOT法表现一致。结论电针预处理可以减轻缺血再灌注损伤后肝细胞凋亡。     

Regulatory Effect of Bcl-2 Family Proteins in CPB-induced Cardiomyocyte Apoptosis in Dog Hearts

Recent studies showed that apoptosis was in-volved in the pathophysiology of various heart dis-eases including congestive heart failure,ischemia-reperfusion injury,allograft,myocarditis,hypother-mic lung preservation and other cardiac pathologies.So far,there have no reports on the myocyte apopto-sis in human hearts following conventional hypother-mic cardiopulmonary bypass,and the mechanismsunderlying myocardiocyte apoptosis are still poorlyunderstood.The aim of the present study was to in-ve…     

补阳还五汤联合依达拉奉对脑缺血再灌注损伤小鼠神经细胞线粒体凋亡途径的影响

目的 研究补阳还五汤联合依达拉奉对脑缺血再灌注损伤的神经保护作用及可能机制。 方法 改良线栓法建立小鼠大脑中动脉缺血再灌注模型。90只小鼠随机分5组(每组18只):假手术组、模型组、补阳还五汤(BYHWD)组(按10 ml/kg灌服给药)、依达拉奉(ED)组(按3 mg/kg尾静脉注射给药)、补阳还五汤+依达拉奉(BYHWD+ED)组(BYHWD按10 ml/kg灌服给药+ED按3 mg/kg尾静脉注射给药)。给药1次/d,连续用药7 d。术后1、3、7 d观察各组神经功能缺损评分。术后7 d,氯化三苯基四氮唑(TTC)染色评估脑梗死体积;TUNEL法检测神经细胞的凋亡情况;免疫组化法观察脑皮质bcl-2、bax、caspase-3表达阳性细胞数。各组数据的比较用单因素方差分析,2组间比较用t检验,P＜0.05为差异有统计学意义。 结果 缺血再灌注后,神经功能评分随时间延长有减少;同一时间点,模型组评分最高,联合用药组评分最低(P＜0.01)。术后7 d各组比较:BYHWD+ED组的脑梗死体积比最小(P＜0.01);各用药组均能降低脑缺血后细胞凋亡数(P＜0.05); bcl-2表达阳性细胞数升高,bax、caspase-3表达阳性细胞数降低,BYHWD+ED组这一变化趋势最明显(P＜0.05)。 结论 两药联用对小鼠脑缺血再灌注损伤有协同神经保护作用,其机制与减少细胞凋亡、调控线粒体凋亡途径蛋白的表达有关。