丁苯酞对血管性痴呆大鼠海马CA1区Bax和Bcl-2表达的影响

目的观察丁苯酞(NBP)对血管性痴呆(VD)大鼠海马CA1区凋亡调控基因Bax和Bcl-2表达的影响,探讨NBP对VD的保护作用。方法将80只健康Wistar大鼠随机分为VD组(VD模型)、NBP治疗组(VD模型+NBP)、NBP对照组(假手术+NBP)、Sham组(假手术组),每组20只,每组又分为4个亚组:术后1、2、4、8 w,每个亚组5只。永久性双侧颈总动脉结扎法制备VD大鼠模型。NBP对照组和NBP治疗组大鼠术后1 d开始给予BNP腹腔注射,剂量为5 mg·kg~(-1)·d~(-1),Sham组和VD组给予生理盐水腹腔注射(0.2 ml/d)。各组大鼠连续腹腔注射给药7 d。术后各时间点(1、2、4、8 w)留取海马组织,免疫组化观察各组大鼠海马CA1区Bax和Bcl-2的表达。结果VD组大鼠海马CA1区Bax蛋白表达灰度明显高于Sham组(P<0.05);4 w和8 w时,NBP治疗组大鼠海马CA1区Bax蛋白表达均明显低于VD组大鼠相应时间点(P<0.05)。8 w时VD大鼠海马CA1区Bax表达灰度明显高于1 w和2 w时(P<0.05)。NBP对照组大鼠海马CA1区Bcl-2表达灰度均明显高于Sham组(P<0.05);8 w时NBP治疗组大鼠海马CA1区Bcl-2表达灰度明显高于VD组(P<0.05)。结论VD大鼠海马CA1区促凋亡蛋白Bax过度表达,抗凋亡蛋白Bcl-2的表达有减弱趋势;NBP对VD大鼠海马CA1区细胞凋亡具有明显的保护作用,可以抑制缺血损伤导致的海马CA1区Bax蛋白过度表达,同时能够提高大鼠海马CA1区Bcl-2蛋白的表达水平,抑制细胞凋亡,促进细胞存活,发挥神经保护作用。     

拮抗或激活促肾上腺皮质激素释放因子受体对肠易激综合征大鼠内脏敏感性及结肠动力的影响

目的 探讨促肾上腺皮质激素释放因子(CRF)及其受体对肠易激综合征大鼠内脏敏感性及结肠动力的影响.方法 SD大鼠60只,随机平均分入空白组(不做处理)、模型组(特殊气味条件刺激和肢体束缚直肠刺激非条件刺激轮替致敏)、干预对照组(造模前侧脑室注射0.9%NaC1)、干预一组(造模前侧脑室注射CRF-R1拮抗剂)和干预二组(造模前侧腑室注射CRF-R2激动剂).采用腹部收缩反射(AWR)评分标准评估各组大鼠肠道敏感性,记录各组大鼠结肠快、慢波波动率、最大振幅、收缩波数及振幅指数等电生理活动改变.采用SPSS16.0统计软件分析,计量资料采用方差分析,等级资料采用秩和检验.结果 以AWR=3分时所需的直肠注水量作为评价指标,模型组大鼠[(0.90±0.11)ml]较空白组[(1.23±0.07)ml]内脏敏感性增高(F＝82.586,P＜0.01);结肠电生理活动增强,造模成功.干预对照组直肠注水量为(0.81±0.11)ml,与模型组[(0.90±0.11)ml]差异无统计学意义(F＝3.734,P＞0.05),干预一组[(1.28±0.07)ml,F＝161.878,P＜0.01]和干预二组[(1.22±0.05)ml,F＝121.564,P＜0.01]较干预对照组内脏敏感性降低.干预对照组大鼠结肠快、慢波波动率、最大振幅、收缩波数及振幅指数等电生理活动与模型组无明显差异(P均＞0.05);干预一组和干预二组大鼠结肠电生理活动均较干预对照组明显减弱(均P＜0.05).结论 CRF在IBS发病中起重要作用,抑制CRF-R1或激活CRF-R2可降低1BS大鼠内脏敏感性并抑制结肠运动.

Abstract：

Objective To explore the effect of corticotropin releasing factor (CRF) and its receptor on visceral sensitivity and colon motility of irritable bowel syndrome (IBS) rats. Methods sixty SD rats were divided randomly and equally into control group (without treatment),model group (sensitized in turn with camphor odor as conditional stimulation and physical restraint in combination with rectal distention pressure as non-conditional stimulation),treatment control group (injected physiological saline into lateral ventricles before treatment),treatment group 1 (injected CRF-R1antagonist into lateral ventricles before treatment),treatment group 2 (injected CRF-R2 agonist into lateral ventricles before treatment). Then the rats' visceral sensitivity were assessed by AWR,and colonic electricity activities such as volatility,maximum amplitude of fast wave and slow wave,interdigestive number of contraction wave and index of contraction were recorded. The data was analyzed with SPSS 16. 0 software. Results By the amount of ractal water injection to reach AWR=3 as the evaluation index,model group [(0. 90±0. 11) ml] showed higher visceral sensitivity than that of control group [(1. 23±0. 07) ml,F=82. 586,P＜0. 01],and colonic electricity activity increased (P＜0. 05),model was successfully set up. There was no significant difference of the amount of ractal water injection between model group [(0. 90±0. 11) ml] and treatment control group [(0. 81±0. 11) ml,F=3. 734,P＞0. 05]. Compared with treatment control group,the visceral sensitivity decreased in treatment group 1 [(1. 28±0. 07) ml,F=161. 878,P＜0. 01] and treatment group 2 [(1. 22±0.05) ml,F=121. 564,P＜0. 01]. There was no significant difference between treatment control group and model group in electricity activities such as volatility,maximum amplitude of fast wave and slow wave,interdigestive number of contraction wave and index of contraction (all P＞0. 05). While the electricity activities was weakened in treatment group 1 and 2 compared with the treatment control group (all P＜0. 05). Conclusions CRF plays an important role in the pathogenesis of IBS. Inhibition of CRF-R1 or activation of CRF-R2 may lower visceral hypersensitivity and decrease colon motility of rats.     

帕金森病运动并发症发生机制中G蛋白偶联受体激酶6与N-甲基-D-天冬氨酸受体关系的研究

目的 研究G蛋白偶联受体激酶6(GRK6)在帕金森病(PD)运动并发症发生机制中与N-甲基-D-天冬氨酸(NMDA)受体的关系.方法 建立PD运动并发症大鼠模型,25只大鼠分为3组.异动症(LID)组10只,腹腔注射左旋多巴甲酯23 d;MK-801处理组10只,第23天左旋多巴甲酯注射前腹腔注射MK-801;PD组5只,腹腔注射0.2%维生素C液.另设假手术组5只为对照组.观察MK-801处理左旋多巴诱导的运动并发症模型大鼠的行为变化,并用免疫组织化学方法和Western印迹方法检测大鼠纹状体区GRK6蛋白的表达情况.结果 PD组大鼠长期使用左旋多巴后出现明显的异常不自主运动,与人类LID具有相似特征.免疫组化结果显示.PD组损伤侧纹状体区GRK6阳性细胞指数减少至(4.81±1.31)×103(P＜0.05),LID组损伤侧GRK6阳性细胞指数进一步减少至(3.23±0.41)×103(P＜0.01).MK-801组LID大鼠异常不自主运动评分减少,药效期延长,同时损伤侧纹状体区GRK6阳性细胞指数增多至(4.64±1.39)×103(P＜0.05).Western印迹法检测结果同免疫组化基本相符,PD组损毁侧/未损毁侧纹状体区GRK6含量比值为(83.7±2.1)%,LID组GRK6值降低为(76.8±2.2)%,而MK-801组GRK6值升高至(91.1±2.7)%(P＜0.01).结论 NMDA受体拮抗剂能逆转大鼠运动并发症的发生,其机制可能与GRK6增多,抑制了谷氨酸受体的过度活化有关.

Abstract：

Objective To investigate the relationship between G protein-coupled receptor kinase 6 (GRK6) and N-methyl-D-aspartate (NMDA) receptor in the mechanism study underlying motor complications in Parkinson's disease (PD).Methods The rat models (n= 25) of Parkinsonism related motor complications were established and were randomly divided into levodopa-induced dyskinesia (LID) group (n= 10,intraperitoneal injection of levodopa for 23 d),MK-801 treatment group (n= 10,intraperitoneal injection of MK-801 at day23 after intraperitoneal injection with levodopa for 22 days) and PD group (n= 5,intraperitoneal injection of vitamin C).Another 5 rats were served as controls (sham-operation group).The behavior changes of rats in MK-801 treatment group were observed,and the expression of GRK6 in the striate of rats was detected by immunohistochemistry and Western blot.Results After the chronic treatment with levodopa methyl ester,PD rats displayed abnormal involuntary movements,which was similar to levodopainduced dyskinesia in PD patients.Immunohistochemistry showed that GRK6-positive cells of lesion side were decreased in LID rats as compared with PD rats [(3.23±0.41 ) × 103 vs.(4.81 ± 1.31 ) ×103,P＜0.01].Rats in MK-801 treatment group displayed the decreased AIM scores and increased peak rotation,and the increased GRK6-positive cells of lesion side as compared with LID rats (P＜0.05).Western blot showed that the levels of GRK6 was 83.7% ±2.1% in PD group (presented as lesion side/unlesion side),76.8% ± 2.2% in LID group and 91.1% ± 2.7% in MK-801 treatment group (intergroups comparison:all P＜0.05).These results were in accordance with the results of immunohistochemistry.Conclusions Antagonist of NMDA receptor can be used to reduce the motor complications in rats.It may be due to increased GRK6 which inhibits the overactivation of glutamic acid receptors.     

普罗布考对2型糖尿病大鼠肾脏硫氧还蛋白系统表达的影响

目的 观察硫氧还蛋白系统在2型糖尿病大鼠肾脏组织的表达及早期普罗布考干预对硫氧还蛋白系统表达的影响.方法 将30只雄性SD大鼠分为正常对照组(C组)、糖尿病组(D组)、糖尿病普罗布考治疗组(P组),普罗布考干预8周后,通过免疫组化、RT-PCR、Western免疫印迹技术观察普罗布考对硫氧还蛋白(Thioredoxin,Trx)及硫氧还蛋白相互作用蛋白(Thioredoxin-interacting protein,Txnip)表达的影响,并检测各组大鼠空腹血糖(FPG)、胰岛素、肾功能、丙二醛、超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)的含量.结果 P组与C组比较,Trx表达明显降低(0.162±0.008对0.239±0.006,P＜0.05),Txnip表达明显升高(0.159±0.003对0.091±0.016,P＜0.05).P组与D组比较,Trx表达升高(0.162±0.008对0.108±0.013,P＜0.05),Txnip表达降低(0.159±0.003对0.236±0.009,P＜0.05).与C组比较,D组大鼠氧化应激指标升高、肾功能受损、抗氧化酶降低,而P组大鼠上述指标(除FPG外)有所改善(P＜0.05).结论 普罗布考通过部分恢复Trx功能,降低Txnip表达,减少氧化应激,发挥对2型糖尿病大鼠肾脏组织的保护作用.

Abstract：

Objective To observe the expression of thioredoxin (Trx) and thioredoxin-interacting protein (Txnip) in the kidney of type 2 diabetic rats induced by streptozotocin and the effect of probucol treatment on thioredoxin system. Methods Thirty male SD rats were divided into control group( C, n = 10), diabetes group ( D, n =10), and probucol treated diabetic group ( P, n = 10). After eight weeks of probucol treatment, the expressions of Trx and Txnip in the kidney of three groups were measured by RT-PCR, Western blot, and immunohistochemistry. Body weight,24 h microalbuminuria( ALB), fasting plasma glucose( FPG), fasting insulin( HNS), blood urea nitrogen (BUN), creatinine (Cr), malondialdehyde ( MDA ), superoxide dismutase ( SOD ), and catalase (CAT) were determined. Results Compared with group C, Trx was markedly decreased in group P (0. 162 ±0. 008 vs 0. 239 ±0. 006, P＜0.05 ), while Txnip was significantly increased (0. 159±0.003 vs 0. 091 ±0.016, P＜0.05 ). Trx in group P was increased as compared with group D (0. 162 ±0. 008 vs 0. 108 ± 0. 013, P ＜ 0. 05 ), while Txnip was lowered (0. 159±0.003 vs 0. 236±0.009 ,P＜0.05 ). FPG, 24 h ALB, BUN, Cr,and MDA levels in group D were markedly increased as compared with group C (P＜0. 05), while the activity of SOD, CAT, and FINS levels were decreased apparently (P＜0.05). The above markers except for FPG in group P were ameliorated (P＜0. 05 ).Conclusions Probucol attenuated oxidative stress by means of partially restoring Trx function and reducing Txnip expression, and thus played a major role in renoprotection of type 2 diabetic nephropathy.     

Effect of replenishment of vitamin D on survival in patients with decompensated liver cirrhosis: A prospective study

AIM To assess the vitamin D(VD) deficiency as a prognostic factor and effect of replenishment of VD on mortality in decompensated cirrhosis. METHODS Patients with decompensated liver cirrhosis were screened for serum VD levels. A total of 101 VD deficient patients( 20 ng/mL) were randomly enrolled in two groups: Treatment group(n = 51) and control group(n = 50). Treatment group received VD treatment in the form of intramuscular cholecalciferol 300000 IU as loading dose and 800 IU/d oral as maintenance dose along with 1000 mg oral calcium supplementation. The VD level, clinical parameters and survival of both the groups were compared for 6-mo.RESULTS Prevalence of vitamin D deficiency(VDD) in decompensated CLD was 84.31%. The mean(SD) age of the patients in the treatment group(M:F: 40:11) and control group(M:F: 37:13) were 46.2(± 14.93) years and 43.28(± 12.53) years, respectively. Baseline mean(CI) VD(ng/mL) in control group and treatment group were 9.15(8.35-9.94) and 9.65(8.63-10.7), respectively. Mean(CI) serum VD level(ng/mL) at 6-mo in control group and treatment group were 9.02(6.88-11.17) and 29(23-35), respectively. Over the period of time the VD, calcium and phosphorus level was improved in treatment group compared to control group. There was nonsignificant trend seen in greater survival(69% vs 64%; P 0.05) and longer survival(155 d vs 141 d; P 0.05) in treatment group compared to control group. VD level had no significant association with mortality(P 0.05). In multivariate analysis, treatment with VD supplement was found significantly(P 0.05; adjusted hazard ratio: 0.48) associated with survival of the patients over 6-mo. CONCLUSION VD deficiency is very common in patients of decompensated CLD. Replenishment of VD may improve survival in patients with decompensated liver cirrhosis.     

追赶生长大鼠胃泌素及内脏脂肪细胞CCK2R表达变化

24只Wistar大鼠分为正常对照组、限食组、追赶生长组,检测所有大鼠血糖、血脂、血清胃泌素,内脏脂肪体脂比、脂肪细胞CCK2R mRNA和蛋白水平.结果 显示限食组和正常组相比,血清胃泌素水平降低54%(P＜0.05),内脏脂肪体脂比减少55%(P＜0.05),脂肪细胞CCK2R mRNA和蛋白表达下降(2.19±0.18对3.2±0.24,0.11±0.03对0.15±0.04,P＜0.05).追赶生长组血清胃泌素水平分别高于限食组72%和正常组31%(P＜0.05),内脏脂肪体脂比高于限食组114%(P＜0.05),达到正常对照组水平;同时脂肪细胞CCK2R mRNA和蛋白表达高于正常对照组(4.09±0.59对3.2±0.24,0.25±0.05对0.15±0.04,P＜0.05).追赶生长大鼠内脏脂肪优先沉积的机制可能和胃泌素分泌增加及脂肪细胞CCK2R表达增加相关.

Abstract：

Wistar rats(n=24) were divided into normal control group(NC), food restriction group(FR), and catch-up group(RN). Serum glucose,lipids, gastrin, the ratio of visceral fat to body fat, adipocyte CCK2R mRNA and protein levels were determined. Compared with NC group, FR rats had lower serum gastrin and visceral fat formation. The adipocyte CCK2R mRNA and protein levels of FR rats were lower than those of NC rats. Serum gastrin level of RN rats was higher than those of FR and NC rats(P＜0.05). The ratio of visceral fat to body fat in RN rats increased compared with FR rats and was close to that of NC rats. The adipocyte CCK2R mRNA and protein levels of RN rats were higher than those of FR and NC rats. Gastrin and its receptor pathway possibly play a role in the mechanism of visceral fat accumulation in catch-up rats.     

Anti-tumor activity of tanshinone ⅡA in combined with cyclophosphamide against Lewis mice with lung cancer

Objective:To explore the anti-tumor activity of tanshinone ⅡA in combined with cyclophosphamide against Lewis mice with lung cancer and the effect on cellular immune function.Methods:Lewis tumor cells were inoculated suhcutaneously into the right armpit of mice in each group(n=20) to establish Lewis lung cancer mice model.After model establishment,mice in the model group were given normal saline by lavage,qd.Mice in treatment Ⅰ group were given intraperitoneal injection of TanIIA,15 mg/kg,qd.Mice in treatment Ⅱ group were given intraperitoneal injection of CTX,25 mg/kg,qd.Mice in treatment Ⅲ group were given intraperitoneal injections of TanIIA and CTX,in which the administration method of TanIIA was the same as in treatment Ⅰ group,continuously for 2 weeks,and the dosage of CTX was the same as in treatment Ⅱ group,24 h after model establishment,every other day.Mice were sacrificed 2 weeks after establishment.The tumor tissues were collected to calculate the anti-tumor rate.Immunohistochemistry was used to detect the expressions of Bcl-2,Bax,VEGF,Angiostatin,and Endostatin.FCM was used to detect T lymphocyte subsets in spleen and liver of mice.Results:The tumor weight in treatment Ⅰ,Ⅱ,and Ⅲ groups was significantly lower than that in the model group(P0.05).The tumor weight in treatment Ⅲ group was significantly lower than that in treatment Ⅰ and Ⅱ groups(P0.05).The anti-tumor rate in treatment Ⅱ and Ⅲ groups was significantly higher than that in treatment Ⅰ group(P0.05).Bcl-2 expression in the tumor tissues of treatment Ⅰ,Ⅱ,and Ⅲgroups was significantly lower than that in the model group(P0.05),while Bax expression was significantly higher than that in the model group(P0.05).Bcl-2 expression in the tumor tissues of treatment Ⅰ and Ⅱ groups was significantly higher than that in treatment Ⅲ group(P0.05),while Bax expression was significantly lower than that in treatment Ⅲ group(P0.05).CD4~+ and CD4~+/CD8~+ in treatment Ⅰ,Ⅱ,and Ⅲ groups were significantly higher than those in the model group(P0.05).CD4~+ in treatment Ⅲ group was significantly higher than that in treatment Ⅰ and Ⅱ groups(P0.05),while CD4~+/CD8~+ was significantly higher than that in treatment Ⅱ group(P0.05).The comparison of CD8~+ among each group was not statistically significant(P0.05).NK cell activity in treatment Ⅰ,Ⅱ,and Ⅲ groups was significantly higher than that in the model group(P0.05).NK cell activity in treatment Ⅲ group was significantly higher than that in treatment Ⅰ and Ⅱ groups(P0.05).Conclusions:TannA in combined with CTX can down regulate Bcl-2 expression in lung cancer tissues,up regulate Bax expression,inhibit the neovascularization of tumor tissues,and enhance the immunological function,with a significant anti-tumor activity.     

Effects of low-calorie diet on steatohepatitis in rats with obesity and hyperlipidemia

AIM: To evaluate the effects of low calorie diet (LCD) on nonalcoholic steatohepatitis (NASH) in rats with obesity and hyperlipidemia.METHODS: 29 Sprague-Dawley (SD) rats were randomly divided into three groups. The animals in control (n=9) and NASH group (n=10) were fed on standard rat diet and high fat diet respectively for 12 weeks, ten rats in LCD group were fed on high fat diet for 10 weeks and then low calorie diet for 2 weeks. At the end of the experiment, body weight, abdominal adipose content, liver function, and hepatopathological changes were examined to evaluate the effect of different feeding protocols on the experimental animals.RESULTS: There was no death of animal in the experimental period. All rats in the NASH group developed steatohepatitis according to liver histological findings. Compared with the control group, body weight (423.5±65.2 vs 351.1±43.0 g,P＜0.05), abdominal adipose content (14.25±1.86 vs9.54±1.43,P＜0.05), liver index (3.784-±0.533 vs2.957±±0.301%, P＜0.01),total serum cholesterol (1.60±0.41 vs 1.27±0.17 mmol/L, P＜0.05)and free fatty acids (728.2±178.5 vs 429.2±96.7 mmol/L,P＜0.01), serum alanine aminotransferase (1 257.51±671.34vs671.34±118.57 nkat/L, P＜0.05) and aspartic aminotransferse (2 760.51±998.66 vs 1 648.29±414.16 nkat/L, P＜0.01) were significantly increased in the NASH group. Whereas, when rats were fed on LCD protocol, their body weight (329.5±38.4 g,P＜0.01), abdominal adipose content (310.21±1.52 g, P＜0.05),liver index (3.199±0.552 %, P＜0.05), and serum alanine aminotransferase (683.03±245.49 nkat/L, P＜0.05) were significantly decreased, and the degree of hepatic steatosis (P＜0.05) was markedly improved compared with those in the NASH group. However, no significant difference was found in serum lipid variables and hepatic inflammatory changes between the two groups.CONCLUSION: LCD might play a role in the prevention and treatment of obesity and hepatic steatosis in SD rats,but it exerts no significant effects on both serum lipid disorders and hepatic inflammatory changes.     

阿托伐他汀对高胆固醇血症小鼠海马中tau蛋白磷酸化水平的影响

Objective To investigate the phosphorylation of tau protein and the effect of atorvastatin on tau protein phosphorylation in hypercholesteremic mouse hippocampus. Methods Male Kunming mice were randomly divided into normal control group, hypercholesteremia group, and low-dose atorvastatin treatment group (8mg·kg-1·d-1), middle-dose group (15mg·kg-1·d-1),and high-dose group (20mg·kg-1·d-1) with five mice in each group. The hypercholesteremia mouse model was induced by cholesterol-enriched diets. The expression level of tau protein phosphorylation in mouse hippocampus was detected by Western blot and immunohistochemistry methods. The activities of mitogen-activated protein kinase (MAPK) and cyelin dependent kinase 5 (Cdk-5) were measured by liquid scintillation counting of the hippocampus incorporated radioactivity and immunoprecipetation activity assay respectively. Results In hypercholesteremic group, the expression level of tau protein phosphorylation in mouse hippocampus was significantly increased(F=14.761,P＜0.01)as compared with control group, and so were MAPK activity and Cdk-5 activity(all P＜0.01). Atorvastatin treatment group showed the decreased expression level of tau protein phosphorylation at ser396/404 site in low-dose group (F=6.349,P＜0.05),middle-dose group (F=10.283,P＜0.01) and high-dose group (F=10.511,P＜0.01) as compared with hypercholesteremia mouse group. The activities of MAPK and Cdk-5 were decreased along with the increasing atorvastatin doses. Conclusions Hypercholesteremia induces tau protein hyperphosphorylation in mouse hippocampus. Abnormal cholesterol metabolism may play an important role in the pathology process of neurodegeneration in brain. Atorvastatin might inhibit tau protein hyperphosphorylation by inhibiting the activations of MAPK and Cdk-5 in brain, atorvastatin may have the protect effect for tau protein.     

尤瑞克林对大鼠脑缺血再灌注损伤Bcl-2和Bax表达的影响

目的 观察大鼠脑缺血再灌注损伤后脑组织中Bcl-2和Bax的表达及尤瑞克林的影响。 方法 48只SD大鼠,随机分为假手术组、模型组、生理盐水对照组和尤瑞克林治疗组。线栓法制作大鼠大脑中动脉脑缺血再灌注模型,缺血2h后再灌注,24 h后行神经功能评分,采用免疫组织化学法和反转录聚合酶链反应(RT-PCR)法观察脑组织Bcl-2和Bax表达的变化。 结果 大鼠脑缺血再灌注损伤后,模型组和生理盐水对照组Bcl-2阳性细胞数和mRNA的表达分别为[(14.28±2.54)个/HP、0.551±0.068和（16.54±1.84）个/HP、0.585±0.084],比假手术组[(7.58±0.97)个/HP、0.324±0.042]增多(P＜0.05);模型组和生理盐水对照组Bax阳性细胞数和mRNA的表达分别为[(24.38±3.58)个/HP、0.540±0.076和(26.74±4.04)个/HP、0.527±0.065],比假手术组[（8.24±1.95）个/HP、0.309±0.037]增多(P＜0.05)。尤瑞克林干预后,Bcl-2阳性细胞数和mRNA的表达显著上调[(25.61±3.41)个/HP、0.791±0.096],Bax阳性细胞数和mRNA的表达下调[(18.54±2.38)个/HP、0.359±0.053],与模型组和生理盐水对照组相比,差异均有统计学意义(P＜0.05)。 结论尤瑞克林可上调脑组织中Bcl-2的表达,下调Bax的表达,从而抑制细胞凋亡,这可能是其发挥脑保护作用的机制之一。     

5种黄酮类化合物对凋亡心肌细胞Bcl-2家族蛋白表达的影响

目的 观察5种黄酮类化合物芦丁(RU)、二氢杨梅素(DMY)、陈皮素(HP)、大豆黄素(DA)、红花黄色素A(HYSA)对H2O2诱导的心肌细胞凋亡的作用,探讨其与克山病的关系及可能的作用机制.方法 培养新生Wistar仔鼠心肌细胞,分为对照组、模型组、药物孵育组,用荧光染色法观察心肌细胞凋亡情况,流式细胞仪检测细胞凋亡率,Western blot法观察凋亡相关基因Bcl-2家族蛋白表达.结果 与模型组[(24.33±6.51)%]比较,RU[(13.95±3.80)%]、DA[(11.82±3.50)%]、HYSA[(12.33±3.78)%]能有效降低心肌细胞凋亡率(P＜0.05).5种黄酮类化合物能不同程度地上调Bcl-2蛋白表达,下调Bax蛋白表达,使Bcl-2/Bax比值增力口[RU(0.989±0.094)、DMY(0.931±0.280)、HP(0.980±0.095)、DA(1.049±0.092)、HYSA(1.031±0.039)],与模型组(0.490±0.046)比较,差异均有统计学意义(P＜0.05);而Bcl-xl表达无明显变化(P＞0.05).结论 RU、DMY、HP、DA、HYSA可以通过Bcl-2、Bax途径调节细胞凋亡过程,发挥抗氧化作用,抑制细胞凋亡,为克山病防治提供了一种思路.     

吡格列酮对高脂血症大鼠主动脉内皮细胞凋亡的作用

目的观察吡格列酮对高脂血症大鼠主动脉内皮细胞凋亡的影响,并探讨其可能的作用机制。方法清洁型SD大鼠26只,随机分为健康对照组(9只)、高脂饮食组(17只),高脂饮食组喂养12周后再随机分为模型组(8只)和吡格列酮组(9只),4周后,检测各组血脂水平,通过免疫组织化学法检测主动脉Bcl-2、Bax的表达,TUNEL染色法观察主动脉内皮细胞凋亡情况,计算细胞凋亡指数。结果 (1)高脂饮食组喂养12周后,血脂明显升高;给药4周后,与模型组比较,吡格列酮组TG、TC水平明显降低(P=0.000);(2)与健康对照组相比,模型组主动脉Bax蛋白表达明显增高(P=0.003),Bcl-2蛋白表达和Bcl-2/Bax比值明显降低(P=0.000);与模型组相比,吡格列酮组主动脉Bax蛋白表达明显降低(P=0.000),Bcl-2蛋白表达(P=0.001)和Bcl-2/Bax比值明显升高(P=0.000),且吡格列酮组主动脉内皮细胞凋亡指数较模型组明显降低,差异有统计学意义(17.5633±7.0584比6.0475±2.2370,P=0.000)。结论吡格列酮可改善高脂血症大鼠血脂水平,调节凋亡蛋白表达,减少主动脉内皮细胞凋亡。     

丹星通络汤治疗大鼠脑缺血再灌注Bax蛋白表达的影响

目的 观察丹星通络汤对大鼠脑缺血再灌注损伤海马区细胞凋亡调控基因Bax蛋白表达的影响,探讨丹星通络汤预防与治疗大鼠脑缺血再灌注损伤的作用.方法 SD大鼠40只,随机分成假手术组、模型组、尼莫地平组、丹星通络汤小剂量组、丹星通络汤大剂量组,每组8只.采用线栓法制备大鼠大脑中动脉闭塞(MCAO)再灌注模型.HE染色检测大鼠脑组织海马区的病理学变化,免疫组织化学法和医学图像分析法检测大鼠脑组织海马区Bax蛋白的平均光密度(OD)值.结果 与模型组比较,丹星通络汤大、小剂量组大鼠脑组织海马区Bax蛋白的OD值明显减低(P＜0.05),并且丹星通络汤大剂量组与尼莫地平组相比有统计学意义(P＜0.05).结论 丹星通络汤可能通过下调Bax蛋白的表达,从而抑制脑组织的凋亡机制发挥对脑组织的保护作用.     

缬沙坦对心力衰竭家兔钙调蛋白依赖性蛋白激酶Ⅱ的影响

目的 探讨家兔慢性心力衰竭(心衰)时心肌钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)蛋白表达及活性的改变及血管紧张素Ⅱ受体拮抗剂缬沙坦长期干预的意义.方法 27只家兔随机分为3组,假手术组、心衰组和缬沙坦组各9只,通过超容量负荷联合压力负荷建立家兔心衰模型,于术后7周观察左心室结构、血液动力学的变化及CaMK Ⅱ的表达和活性的改变.结果 与假手术组比较,心衰组左室重量指数(LVMI)、左窒舒张末压显著升高(P＜0.05),左室短轴缩短率及左室射血分数明显降低(P＜0.05);与心衰组比较,缬沙坦组左室重量指数、左室舒张未压显著降低(P＜0.05),左室短轴缩短率及左室射血分数明显升高(P＜0.05);心衰组CaMK Ⅱ蛋白表达及活性显著高于假手术组(P＜0.05);缬沙坦组CaMKⅡ蛋白表达及活性显著低于心衰组(P＜0.05).结论 缬沙坦长期干预心衰,能够改善心脏舒缩功能,可能与其降低CaMK Ⅱ蛋白表达及活性有关.     

着色性干皮病基因B对白介素-6介导的血管平滑肌细胞增殖及凋亡的影响

目的 探讨着色性干皮病基因B(xeroderma pigmentosum B,XPB)对白介素-6(IL-6)介导人血管平滑肌细胞( VSMC)增殖及凋亡的影响.方法 用脂质体转染法将重组质粒pcDNA3.1-XPB和空载质粒pcDNA3.1稳定转染VSMC,然后给予100 U/ml的IL-6孵育48 h.实验分为6组:(1)空白对照组;(2) pcDNA3.1组;(3) pcDNA3.1-XPB组;(4)IL-6组;(5) IL-6+ pcDNA3.1组;(6)IL-6+ pcDNA3.1-XPB组.用反转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法(Western blotting)检测XPB、Bcl-2、Bax和野生型p53(wt-p53)表达量的变化;用比色法(MTT)观察细胞增殖活力;用流式细胞仪检测细胞周期和凋亡率.结果 重组质粒pcDNA3.1-XPB转染使细胞XPB表达增高(P＜0.05或P＜0.01),同时Bcl-2表达降低、Bax和wt-p53表达增高(P＜0.05或P＜0.01),抑制IL-6促进VSMC的Bcl-2高表达、Bax和wt-p53降表达(P＜0.05或P＜0.01);XPB高表达抑制了细胞增殖活力(q=2.95,P＜0.05),并抑制IL-6促进VSMC增殖,IL-6+ pcDNA3.1-XPB组和IL-6 +pcDNA3.1组VSMC存活率分别为(102.6±6.2)％和(124.5+7.9)％(q=3.49,P＜0.05);流式细胞仪检测结果显示,XPB高表达引起细胞G0/G1期增加、S期减少、凋亡率增加(q值分别为2.99、5.39、3.05,P＜0.05或P＜0.01),并抑制IL-6促进VSMC G0/G1期减少、S期增加、凋亡率降低的作用,IL-6+ pcDNA3.1-XPB组和IL-6 +pcDNA3.1组分别为(70.9±6.7)％与(54.8±2.9)％、(20.2±3.6)％与(36.4±7.2)％、(5.9±2.1)％与(0.3±0.1)％(q值分别为6.91、8.54、7.53,均P＜0.01).结论 XPB基因能抑制VSMC增殖并促进其凋亡,并能抑制IL-6促进VSMC增殖和降低其凋亡的作用,有望成为治疗动脉粥样硬化的靶点.     

白藜芦醇对慢性脑低灌注大鼠认知功能损害的保护作用

目的 观察白藜芦醇对慢性脑低灌注大鼠认知功能损害的影响,并探讨白藜芦醇抗氧化机制在其中的作用.方法 采用永久性双侧颈总动脉结扎术(2VO)制备慢性脑低灌注大鼠模型,健康雄性Wistar大鼠随机分为假手术组、2VO组、2VO+白藜芦醇组.通过Morris水迷宫观察大鼠的空间学习及记忆功能,免疫组化检测皮质及海马CA1区4-羟基壬烯醛(4-hydroxynonenal,4-HNE)和8-羟化脱氧鸟苷(8-hydroxy-2′-deoxyguanosine,8-OHdG)表达的变化,以反应脂质和DNA的氧化性损伤.结果 2VO组大鼠第3～5天逃逸潜伏期分别为(42.1±5.4)s、(36.4±4.4)s、(30.4±4.0)s,与假手术组大鼠(25.1±3.3)s、(12.4±3.3)s、(8.1±3.4)s相比,逃逸潜伏期明显延长(q=10.91、14.54、14.07,均P＜0.01),2VO组大鼠目标象限探索时间(12.9±2.5)s,明显少于假手术组(18.9±2.2)s(q=6.47,P＜0.01);2VO+白藜芦醇组大鼠第3～5天逃逸潜伏期分别为(29.5±4.0)s、(25.6±4.3)s、(19.8±4.2)s,与2VO组大鼠(42.1±5.4)s、(36.4±4.4)s、(30.4±4.0)s比较,逃逸潜伏期缩短(q=7.71、6.22、6.37,均P＜0.01),2VO+白藜芦醇组大鼠目标象限探索时间(16.5±1.8)s,比2VO组大鼠(12.9±2.5)s延长(q=3.83,P＜0.05).2VO组大鼠皮质与海马CA1区的8-OHdG平均积分光密度值(IOD)分别为265.1±9.0和37.8±5.0,与假手术组168.2±6.0和24.0±4.0相比均增加(q=31.89、7.48,均P＜0.01);2VO+白藜芦醇组与2VO组相比,皮质与海马CA1区的8-OHdG平均IOD均降低,分别为195.1±7.0和26.0±4.3(q=23.03、6.49,均P＜0.01).结论 白藜芦醇能减轻大鼠慢性脑低灌注所致的血管性认知功能损害,其作用可能与抑制慢性脑低灌注后的氧化性损伤有关.     

国产与进口西罗莫司洗脱支架置入后血管内超声随访的对比

目的 利用血管内超声对比观察国产与进口西罗莫司洗脱支架对冠心病患者支架术后新生内膜增生的抑制作用.方法 2003年5月至2007年3月,对215例冠心病患者(317处病变)置入西罗莫司洗脱支架,并在术后1年行冠状动脉造影和血管内超声(IVUS)检查.其中Firebird组108例患者(147处病变)置入国产西罗莫司洗脱支架(Firebird支架),Cypher组107例患者(138处病变)置入进121西罗莫司洗脱支架(Cypher支架).结果 两组患者一般临床情况差异无统计学意义.两组靶病变部位、病变长度、狭窄程度及病变类型差异均无统计学意义,但Firebird组术后最小管腔直径大于Cypher组[(2.88±0.43)mm比(2.78±0.33)mm,P＜0.05].随访定量冠状动脉造影分析显示,Firebird组与Cypher组支架内晚期管腔丢失[(0.17±0.29)mm比(0.16±0.27)mm,P＞0.05]和节段内晚期管腔丢失[(0.18±0.36)mm比(0.20±0.32)mm,P＞0.05]差异均无统计学意义.IVUS分析显示,与Cypher组比较,尽管Firebird组支架面积[(6.99±2.25)mm~2比(6.46±1.71)mm~2,P＜0.05]、管腔面积[(6.89±2.30)nm~2比(6.36±1.73)mm~2,P＜0.05]、支架体积[(162.5±68.9)mm~3比(140.8±57.9)mm~3,P＜0.01]、管腔体积[(160.4±69.5)mm~3比(138.6±57.6)mm~3,P＜0.01]及最小支架面积[(5.40±1.85)mm~2比(4.92±1.43)mm~2,P＜0.05]均较大,但两组的内膜增生容积[(2.09±5.46)mm~3比(2.23±6.50)mm~3,P＞0.05]和内膜增生容积百分数[(1.68±5.84)%比(1.59±4.10)%,P＞0.05]差异均无统计学意义.结论 Firebird支架置人后再狭窄的发生率较低,抑制内膜增生作用与Cypher支架相似.     

Protective effect of RadixAstragali injection on immune organs of rats with obstructive jaundice and its mechanism

AIM： To observe the protective effect of RadixAstragali injection on immune organs （lymph nodes, spleen and thymus） of rats with obstructive jaundice （OJ） and its mechanism.
METHODS： SD rats were randomly divided into sham-operation group, model control group and Radix Astragali treatment group. On days 7, 14, 21 and 28 after operation, mortality rate of rats, pathological changes in immune organs, expression levels of Bax and nuclear factor （NF）-κB p65 proteins, apoptosis indexes and serum tumor necrosis factor （TNF）-α level in spleen and thymus were observed, respectively.
RESULTS： Compared to model control group, the number of dead OJ rats in Radix Astragali treatment group decreased （P 〉 0.05）. The TNF-α level （27.62 ± 12.61 vs 29.55± 18.02, 24.61 ± 9.09 vs 31.52± 10.95） on days 7 and 21, the pathological severity score for spleen [0.0 （0.0） vs 0.0 （2.0） on days 7 and 14 and for lymph nodes [0.0 （1.0） vs 1.0 （2.0）, 1.0 （0.0） vs 2.0 （1.0）] on days 21 and 28, the product staining intensity and positive rate of Bax protein in spleen [0.0 （0.0） vs 1.0 （2.0）, 0.0 （1.0） vs 2.0 （1.5） and thymus [0.0 （0.0） vs 1.0 （2.0）, 0.0 （1.0） vs 2.0 （1.5）] on days 14 and 28, the apoptotic indexes [0.0 （0.0） vs 0.0 （0.01）] in spleen and thymus [0.0 （0.0） vs 0.0 （0.01） on days 14 and 21 were significantly lower in Radix Astragali treatment group than in model control group （P 〈 0.05）.
CONCLUSION： Radix Astragali has protective effects on immune organs of OJ rats by relieving the pathological changes in immune organs, reducing TNF-α level and inhibiting Bax expression and apoptosis in spleen and thymus.     

八面蒙脱石联合莫沙必利灌胃对急性坏死性胰腺炎大鼠回肠NF-κB活化的影响

目的 探讨八面蒙脱石对急性坏死性胰腺炎(ANP)大鼠回肠黏膜NF-κB活化的影响.方法 120只雄性SD大鼠按完全随机法分为假手术组、ANP组、莫沙必利组、八面蒙脱石组和莫沙比利+八面蒙脱石(联合)组,每组24只大鼠.采用胰胆管内逆行注射牛磺胆酸钠诱导大鼠ANP模型;假手术组开腹注射生理盐水后关腹.各治疗组于造模前30 min分别给予相应药物灌胃1次.术后3、6、12 h分批处死动物.取胰腺组织行病理检查;取血测定TNF-a、IL-1水平;取回肠观察超微结构的改变;免疫组化法检测回肠黏膜NF-κB的表达;Western blotting法检测回肠组织IκBa水平.结果 与假手术组比较,ANP组胰腺损伤明显,血TNF-a、IL-1水平明显升高[(36.79±1.14)pg/ml对(9.00±0.01)pg/ml,(25.17±1.93)ng/ml对(3.45±0.11)ng/ml,P值均＜0.05],回肠绒毛高度显著下降[(0.25±0.22)×10~(-6)m对(1.50±0.33)×10~(-6)m,P＜0.05],柱状细胞指数显著降低[(2.40±1.65)×10~6个/m对(13.5±4.28)×10~6个/m,P＜0.05],NF-κB表达增加(6.92±1.54对1.28±0.29,P＜0.05),IκBa蛋白水平下降(3.30±1.99对24.32±1.93).莫沙必利组和八面蒙脱石组的各项指标与ANP组均无显著差异.联合组的血TNF-a、IL-1水平[(30.57±0.39)pg/ml和(13.45±1.26)ng/ml]、回肠绒毛高度和柱状细胞指数[(1.05±0.28)×10~(-6)m和(10.10±2.50)×10~6个/m]、NF-κB活化(4.32±1.57)和IκBa蛋白水平(15.99±1.26)均与ANP组有统计学差异(P值均＜0.05).结论 八面蒙脱石联合莫沙必利灌胃能减少ANP大鼠回肠黏膜NF-κB的大量激活,减轻胰腺和回肠黏膜的损伤程度.