转基因诱导人血红素加氧酶1过表达对大鼠肾脏缺血再灌注损伤的保护作用

目的 观察转基因方法诱导人血红素加氧酶1(hHO-1)过表达对大鼠肾脏缺血再灌注损伤的作用.方法 构建SD大鼠的肾脏缺血再灌注模型.治疗组(18只),经供肾动脉用负载人血红素加氧酶-1腺病毒(Ad-hHO-1)2.5×109pfu/只灌注保存1 h用以转染肾细胞.模型组(12只)和空载体组(18只)则分别用生理盐水1.0 ml和腺病毒空载体(Ad-EGFP)2.5×109pfu/只代替.检测各组大鼠外周血尿素氮和肌酐水平;HE染色和电镜观察缺血再灌注损伤后肾脏的组织形态变化;原位末端标记(TUNEL)法检测肾组织中细胞凋亡情况;免疫组化方法观察肾脏内hHO-1的表达;定量检测肾组织中hHO-1的酶活性.结果 经肾动脉灌注Ad-hHO-1可以成功转染大鼠肾脏细胞,治疗组肾组织中hHO-1的酶活性[(1.62±0.07)nmol·mg-1·min-1]明显高于模型组[(1.27±0.07)nmol·mg-1·min-1]和空载体组[(1.22±0.06)nmol·mg-1·min-1](P＜0.01).免疫组化证实治疗组的肾组织内有大量hHO-1表达,而凋亡细胞减少,相应的肾功能生化检查及肾组织形态变化较模型组和空载体组明显改善.结论 hHO-1在大鼠肾脏中的过表达可以显著减弱缺血再灌注对肾脏引起的损伤.     

乌司他丁对出血坏死性胰腺炎兔心肌缺血/再灌注损伤的保护作用

目的 通过观察出血坏死性胰腺炎兔心肌缺血/再灌注、超氧化物歧化酶(SOD)和丙二醛(MDA)及心肌细胞凋亡的变化,探讨乌司他丁预处理对心肌缺血/再灌注损伤的保护作用.方法 逆行胰管注射5%牛磺胆酸钠溶液制作急性坏死性胰腺炎(ANP)模型.ANP兔随机分为缺血/再灌注生理盐水治疗组(C组,12只); 缺血/再灌注乌司他丁治疗组,该组共分两个亚组(U1组,12只;U2组,12只),U1组在阻断前经静脉注射乌司他丁1万U/kg;U2组在阻断前经静脉注射乌司他丁2万U/kg.每组兔均接受左冠状动脉前降支阻断60 min和再灌注180 min.分别于阻断前(T0)、阻断60 min(T1)、再灌注60 min(T2)、120 min(T3)、180 min(T4)测定血浆SOD、MDA浓度.于T4时点后立即将兔处死,TUNEL法观察兔缺血/再灌注心肌细胞凋亡的变化.结果 再灌注后180 min,U2组血浆MDA T1、T2、 T3、T4明显低于C组(P<0.05);U1组T1、T2明显低于C组(P<0.05);各组血浆SOD自T0后不同程度下降,U2组明显升高,U2组T3、T4与C组比较差异有统计学意义(P<0.05);乌司他丁治疗组心肌凋亡与C组相比明显减少(P<0.01).结论 乌司他丁对出血坏死性胰腺炎兔心肌缺血再灌注损伤有保护作用且高剂量组更为明显. Abstract： Objective To study the effects of ulinastain preconditioning on ischemia-reperfusion injury of myocardium in rabbit with severe acute pancreatitis. Methods The mode of ANP was established by retrograte injection of 5% sodium taurocholate solution into the biliopancreatic duct in the rabbits. In the ANP model, Thirty-six healthy male rabbits weighed 2-2.5 kg were randomly allocated to 3 groups and 12 animals each group: The controlling group were treated with saline(group C); and group U1 and group U2 were pretreated with ulinastain(10 000 U/kg) and ulinastain(20 000 U/kg). All the rabbits in the three groups were subjected to one-hour occlusion of left anterior descending coronary artery and then three-hours reperfusion. Plasma concentrations of SOD and MDA in blood samples were measured at T0 (before occlusion), T1 (60 min after occlusion), T2(60 min after reperfusion),T3(120 min after reperfusion), T4(180 min after reperfusion),respectively. At T4, after killing all rabbits, the TUNEL method was used to examine the changing of myocardium apoptosis against ischemia-reperfusion injury. Results The plasma concentrations of MDA in group U2 at T1,T2,T3,T4 were much lower than that in group C(P＜0.05); The plasma concentrations of SOD in group U2 at T3,T4 were much higher than that in group C (P＜0.05). The apoptosis rate in group U1 was(20±6)%, group U2 was (10±3)% was lower than that in group C[(35±5)%]significantly (P<0.01). Conclusions Ulinastain preconditioning has protective effects against ischemia-reperfusion injury in rabbit with severe acute pancreatitis, especially obvious for high dose.     

针刺预处理对高龄大鼠短暂性脑缺血/再灌注损伤后海马神经元的保护作用

目的 观察针刺预处理对老年大鼠短暂性缺血/再灌注损伤后海马CA1区神经元凋亡的影响.方法 健康老年雄性Wistar大鼠120只,随机分为四组每组30只.组Ⅰ(单纯脑缺血/再灌注组):四动脉阻断法全脑缺血4 min建立老年大鼠全脑缺血模型;组Ⅱ(针刺预处理后脑缺血/再灌注组):连续5 d对百会穴实施穴位针刺预处理(疏密波,强度1mA,频率15 Hz),每天持续30 min,然后建立伞脑缺血模型;组Ⅲ(单纯针刺预处理组):仅针刺百会穴30 min/d,持续5 d;组Ⅳ(假手术组),仪暴露4条动脉,不缺血.每组分别于缺血后12 h、1、2、3和7 d各随机处死6只大鼠,随机抽取一只动物提取海马CA1区脑组织作透射电镜观察神经元超微结构,余下的5只光镜下观察海马神经元形态和凋亡细胞,监测Caspage-3蛋白表达.结果 各组于光镜和电镜下均可见凋亡神经元,与针刺对照组(再灌注2 d时6.7个±0.8个)和假手术组(再灌注2 d时7.2个±0.7个)相比,脑缺血再灌注组和针刺预处理后脑缺血再灌注组细胞凋亡数和Caspase-3表达阳性细胞数均显著增多,再灌注2 d后达峰值,差异有统计学意义(P<0.01);与脑缺血再灌注组(再灌注2 d时44.5±2.0比78.8±4.5)相比,针刺预处理后脑缺血再灌注组细胞凋亡数(冉灌注2 d时30.6±2.7个)和Cagpage-3表达阳性细胞数(再灌注2 d时43.8±2.4个)显著减少,差异有统计学意义(P<0.01).结论 针刺预处理可通过调控caspase-3蛋白的表达对老年大鼠短暂脑缺血再灌注损伤起到的保护作用.

Abstract：

Objective To investigate the effects of acupuncture pre-conditioning on apoptosis in hippocampal neurons following ischemia-reperfusion injury in aged rats. Methods A total of 120 senile male Wistar rats aged 19-21 months (corresponding to 60-year-old human being) weighting 550-710 g were randomly divided into 4 groups (n = 30 each). Cerebral ischemie group: 4-vessel-block was conducted for 4 minutes to establish cerebral ischemie models; Acupuncture pre-eonditioning group: electroacupuneture was applied at acupoint Baihui ( GV20 ) with a frequency of 15 Hz and 2 mA for 30 minutes once daily for 5 days. Then the rats received 4-vessel-block for 4 minutes; Sham-operation group: 4 vessels were exposed; Sham-acupuncture group: only eleetroaeupuncture for 5 days without operation. The rats were sacrificed at the end of predetermined duration of reperfusion 12 h, 1, 2, 3 and 7 d respectively. The brains were immediately harvested and hippocampal CA1 region was isolated for ( 1 ) light and electron microscopic examinations of hippoeampal neurons; (2) detection of apoptotic neurons (TUNEL) ; (3) determination of easpase-3 protein expression with SABC (streptavidin-biotin-poroxidase complex) immuno-histoehemieal technique. Results There were apoptotic neurons in all groups. The numbers of apoptotic neurons and positive neurons of caspase-3 significantly increased in the acupuncture pre-conditioning and cerebral isehemic groups versus the sham-acupuncture and sham-operation groups ( P < 0. O1 ). And the numbers of apoptotic neurons and positive neurons of caspase-3 significantly decreased in the acupuncture preconditioning group versus the cerebral ischemic group (P < 0.01 ). Conclusion Acupuncture preconditioning can decrease the neuronal apoptosis after ischemia-reperfusion injury through a lowered expression of caspase-3 protein in senile rats.     

盐酸戊乙奎醚对肢体缺血再灌注大鼠肾脏的保护作用

目的 探讨盐酸戊乙奎醚后处理对肢体缺血再灌注后肾脏的保护作用及其机制.方法 健康成年雄性Wistar大鼠72只,体重220～250 g,随机数字表法分为3组:对照组(C组)、肢体缺血再灌注组(I/R组)、盐酸戊乙奎醚后处理组(P组),根据缺血后再灌注时间点各组又分为缺血3 h(T0)、再灌注1 h(T1)、3 h(T2)、6 h(T3)四个亚组(n=6).除C组外,各组在大鼠双后肢根部用橡皮筋结扎,完全阻断血流3 h.P组在再灌注前3 min尾静脉注射盐酸戊乙奎醚0.15 mg/kg(0.8ml).比色法检测血清肌酐(Cr)和尿素氮(BUN)的含量、肾脏组织超氧化物歧化酶(SOD)的活性及丙二醛(MDA)的含量,酶联免疫吸附法测定血清肿瘤坏死因子α(TNF-α)的含量,免疫组织化学SABC法测定肾脏组织中缺氧诱导因子1α(HIF-1α)的表达,光镜下观察肾脏组织的病理学改变.结果 I/R组和P组血清BUN、Cr水平、SOD的活性、MDA水平、TNF-α水平、HIF-1α的表达均高于C组(均P<0.05);P组血清BUN、Cr、MDA、TNF-α水平及、HIF-1α表达均低于I/R组[T2时间点:(15.10 ±1.88)mmol/L比(19.46±2.76)mmol/L、(113±10)μmol/L比(143±11)μmol/L、(13.8 ±1.7)nmol/g比(15.5±1.8)nmol/g、(53.1±3.1)ng/L比(53.9±4.8)ng/L、0.298±0.015比0.471±0.032,均P<0.05],SOD的活性高于I/R组(P<0.05).结论 盐酸戊乙奎醚后处理可以下调HIF-1α的表达,减轻肢体缺血再灌注后肾脏的损伤.其机制可能是抑制了炎症反应及氧自由基的释放,改善了肾脏组织的缺血、缺氧状态.

Abstract：

Objective To evaluate the protection of penehyclidine hydrochloric postconditioning on HIF-1α (hypoxia-inducible factor -1α) in renal tissue injury induced by lower limb ischemia/reperfusion (I/R). Methods A total of 72 adult male Wistar rats weighing 230 - 250 g were randomly divided into 3 groups: control ( group C ) , limb ischemia-reperfusion ( group R/I) and penehyclidine hydrochloride postconditioning (group P). The animals were anesthetized by inhaling 2% isoflurane and blood flow of bilateral lower limbs was blocked with rubber bands for 3 h in groups P and R/I. In group P, penehyclidine hydrochloride 0. 15 mg/kg was injected via caudal vein at 3 min pre-reperfusion. After sacrificing, their kidneys were removed at 3 h of ischemia and 1, 3, 6 h of reperfusion respectively. The blood urea nitrogen (BUN) and creatinine ( Cr) were detected by colorimetric method, plasma tumor necrosis factor-α (TNF-α) by ELISA ( enzyme-linked immunosorbent assay ) and HIF-1α of renal tissue by immunohistochemistry. Renal pathological changes were observed under light microscope. Results Compared with group C, the serum levels of BUN and Cr increased while TNF-α and HIF-1α were upregulated in groups I/R and P (P < 0. 05). As compared with group I/R, the serum levels of BUN, Cr and MDA decreased while TNF-α and HIF-lα were down-regulated in group P . [at T2: (15. 10 ± 1. 88) mmol/L vs(19.46±2. 76) mmol/L, (113 ±10) μmol/L vs(143 ± 11) μmol/L, (13. 8 ±1.7) nmol/g vs (15.5 ±1.8) nmol/g, (53.1 ±3. 1)ng/L vs(53.9 ±4. 8) ng/L, 0.298 ±0.015 vs 0.471 ±0.032, all P<0.05 ]. Conclusion Penehyclidine hydrochloride can down-regulate the expression of HIF-lα and attenuate the renal injury induced by lower limb I/R. And the mechanisms may be through inhibiting the inflammatory reactions, reducing the release of oxygen free radicals and improving the conditions of hypoxia and ischemia.     

Effects of testosterone on norepinephrine release in isolated rat heart

The effects of testosterone on norepinephrine release were investigated in the isolated rat hearts.Sprague-Dawley male rats (n=120) were randomized to testosterone and control groups.The rats in testosterone group were perfused with modified Krebs-Henseleit buffer containing different concentrations of testosterone (0.1,1.0,10.0,and 100.0 nmol/L,respectively).Myocardial ischemia was induced by globally stopping the perfusion flow.Exocytotic norepinephrine release was induced by electrical field stimulation at 5 V (effective voltage) and 6 Hz (pulse width of 2 ms) for 1 min.The overflow of norepinephrine was determined by high pressure liquid chromatography and electrochemical detec-tion (HPLC-EC).Following acute ischemia,testosterone (1.0,10.0 and 100.0 nmol/L) significantly re-duced norepinephrine release (P<0.01),and the norepinepherine overflow was similar between the con-trol and 0.1 nmol/L testosterone group (P>0.05).Electrical stimulation of the ventricle evoked norepi-nepherine release,and this was diminished by the perfusion with testosterone at the concentrations of 1.0,10.0 and 100.0 nmol/L (P<0.01).It is suggested that testosterone suppresses ischemia-and electri-cal stimulationinduced norepinepherine release in the isolated rat hearts.     

七氟烷后处理通过调节Bcl-2家族蛋白表达减轻离体大鼠心肌细胞凋亡

目的 研究七氟烷后处理对离体大鼠心肌细胞凋亡蛋白表达的影响.方法 雄性SD大鼠30只,体重230～250 g,使用随机数字表法将实验动物随机分为3组(n=10):假手术组,缺血再灌注组(I/R组),七氟烷后处理组(SPC组).采用Langendroff离体心脏灌注模型,除假手术组外其余两组平衡灌注30 min,全心缺血40min,再灌注120 min.SPC组在再灌注即刻给予2.5%(约1.0MAC)七氟烷10 min后改用普通Krebs-Henseleit(K-H)液灌注.记录平衡灌注末,再灌注30、60、90、120min时左室收缩压(LVSP)、左室舒张末压(LVEDP)、左室发展压(LVDP)、左室内压上升最大速率(+dp/dt)、左室内压下降最大速率(-dp/dt)、心率(HR)以及冠脉流量(CF).分光光度法测量肌酸激酶(CK)和乳酸脱氢酶(LDH).再灌注末TTC染色法测定心肌梗死面积.Western印迹法测定凋亡蛋白Bcl-2、Bax表达情况.结果 SPC组LVSP、LVDP、±dp/dt和CF在再灌注30 min后各时点均高于I/R组(均P＜0.05),LVEDP均低于I/R组(均P＜0.05);HR较I/R组高,但差异无统计学意义(P＞0.05).SPC组LDH和CK释放量均低于I/R组(均P＜0.05),梗死面积低于I/R组(22.2%±2.8%比44.9%±6.6%,P＜0.05).与I/R组比较,SPC组Bcl-2表达高,Bax表达低(均P＜0.05).结论 七氟烷后处理可改善心功能,降低心肌梗死面积,通过调节凋亡蛋白表达减轻离体大鼠心肌细胞凋亡.

Abstract：

Objective To explore the effects of sevoflurane postconditioning on ischemic/reperfused myocardial apoptosis.Methods Isolated perfused rat hearts were randomly assigned into 3 groups:shamoperation ( sham), ischemia/reperfusion (I/R) and sevoflurane postconditioning (SPC).Except for the sham group, the hearts were subjected to 40 min global myocardial ischemia and 120 min reperfusion.Left ventricular systolic pressure (LVSP), left ventricular developed pressure (LVDP), left ventricular enddiastolic pressure ( LVEDP), maximum increase rate of LVDP ( + dp/dt), maximum decrease rate of LVDP ( - dp/dt), heart rate (HR) and coronary flow (CF) were measured at baseline, R (reperfusion) 30 min, R60 min, R90 ain and R120 min.Creatine kinase (CK) and lactate dehydrogenase (LDH) were measured at 5 min and 10 min post-reperfusion.Infarct size was determined by triphenyltetrazolium chloride staining at the end of reperfusion.The expressions of Bcl-2 and Bax were determined by Western blot.Results The values of LVSP, LVDP, ± dp/dt and CF were higher while that of LVEDP was lower in the SPC group than the I/R group at all time points of reperfusion ( P ＜ 0.05 ).The releases of CK and LDH and infarct size were significantly reduced in the SPC group versus the I/R group (22.2% ±2.8% vs L/R:44.9% ±6.6%, P ＜0.05).The expression of Bcl-2 increased significantly while that of Bax decreased in the SPC group verus the I/R group.Conclusion Sevoflurane postconditioning may improve myocardial functions, reduce infarct size and attenuate myocardial apoptosis.And the modulated expression of apoptotic proteins plays an important role in sevoflurane-induced myocardial protection.     

Ischemic preconditioning in immature hearts： mechanism and compatibility with cardioplegia

Objective To investigate (1) whether ischemic preconditioning (IPC) could protect immature rabbit hearts against ischemia-reperfusion injury and (2) the role of K(ATP) channel in the mechanism of myocardial protection. Since cardioplegia is a traditional and effective cardioprotective measure in clinic, our study is also designed to probe the compatibility between IPC and cardioplegia. Methods New Zealand rabbits aged 14-21 days weighing 220-280 g were used. The animals were anesthetized and heparinized. The chest was opened and the heart was quickly removed for connection of the aorta via Langendorff’s method within 30 s after excision. All hearts were perfused with Krebs-Henseleit buffer balanced with gas mixture (O［２］∶CO(2)=95%∶5%) at 60 cm H［2O］ (perfusion pressure). IPC consisted of 5 min global ischemia plus 10 min reperfusion. Glibenclamide was used as the K(ATP) channel blocker at a concentration of 10 μmol/L before IPC. Cardiac arrest was induced with 4℃ St. Thomas cardioplegic solution, at which point the heart was made globally ischemic by withholding perfusion for 45 min followed by 40 min reperfusion. Thirty immature rabbit hearts were randomly divided into four groups: CON (n=9) was subjected to ischemia-reperfusion only; IPC (n=9) underwent IPC and ischemia-reperfusion; Gli (n=6) was given glibenclamide and ischemia-reperfusion; and Gli+IPC (n=6) underwent glibenclamide, IPC and ischemia-reperfusion. Coronary flow (CF), HR, left ventricle developed pressure (LVDP), and ±dp/dt(max) were monitored at equilibration (baseline value) and 5, 10, 20, 30 and 40 min after reperfusion. The values resulting from reperfusion were expressed as a percentage of their baseline values. Arrhythmia quantification, myocardial enzyme in the coronary effluent and myocardial energy metabolism were also determined. Results The recovery of CF, HR, LVDP and ±dp/dt(max) in preconditioned hearts was best among the four groups. The incidence of arrhythmia was low and less CK-MB leaked out in the IPC group. Myocardial ATP content was better preserved by IPC. Pretreatment with glibenclamide completely abolished the myocardial protection provided by IPC, but did not affect ischemia-reperfusion injury. Conclusions While applying cardioplegia, IPC provides significant cardioprotective effects. Activation of K(ATP) channels is involved in the mechanism of IPC-produced cardioprotection.     

Astragaloside IV Regulates Expression of ATP-sensitive Potassium Channel Subunits after Ischemia-reperfusion in Rat Ventricular Cardiomyocytes

Objective: Astragaloside IV (AsIV) is the major effective component extracted from the Chinese herb Astragalus membranaceus, which has been widely used to treat cardiovascular disease. Recent studies have shown that AsIV can potentially protect the heart from myocardial ischemic injury, but the mechanisms of action are unknown. ATP-sensitive potassium (KATP) channels are activated during ischemia and exert a compensatory protective effect on cardiomyocytes. We therefore examined the effects of AsIV on KATP channel currents and channel expression in isolated rat ventricular cardiomyocytes after ischemia-reperfusion injury. Methods: Forty Wistar rats were divided into five groups: control group, ischemia-reperfusion (IP) group, IP + glibenclamide group, IP + pinacidil group and IP + AsIV group. The ischemia-reperfusion injury model was established in enzymatically isolated ventricular cardiomyocytes by perfusion with calcium-free Tyrode solution for 10 min, arrest for 30 min, and reperfusion for 45 min. The different drugs were applied for 10-15 min, and the KATP channel current (IKATP) was recorded with voltage-clamp mode by whole-cell patch-clamp technique. Protein and mRNA expression of the KATP channel subunits Kir6.1, Kir6.2, SUR2A and SUR2B was quantified using western blotting and real-time PCR. Results: The KATP current in IP group was significantly greater than that in control group (211.45±33.67 vs 83.51±23.67 pA; P<0.01). Glibenclamide (10 μmol/L) blocked KATP currents, whereas both AsIV (1 mg/L) and the known channel opener pinacidil (50 μmol/L) significantly increased IKATP (P<0.05). Consistent with this, AsIV significantly up-regulated protein and mRNA expression of Kir6.1, Kir6.2, SUR2A, SUR2B (P<0.01 vs IP group). Conclusion: The protective effects of AsIV in ischemia-reperfusion injury may be related to the up-regulation of several KATP channel subunits and facilitation of KATP currents.     

CD59 prevents human complement-mediated injuries in isolated guinea pig hearts

Objective To assess complement-mediated myocardial injury on isolated guinea pig working hearts and cardioprotective effects of CD59. Methods Using a modified Langendorff apparatus, isolated guinea-pig working hearts were perfused with a modified Krebs Henseleit buffer containing 3% heat-inactivated human plasma and zymosan （IPZ） （control） (n=10), 3% normal human plasma and zymosan （NPZ） (n=10), or 3% normal human plasma and zymosan and 1.5μg/ml CD59 （NPZC）(n=10), respectively. Epicardial electrocardiogram (ECG), cardiac output (CO), coronary arterial flow (CF), maximum left ventricular developed pressure (LVP(max)), maximum left ventricular developed pressure increase rate (+dp/dt(max)), maximum left ventricular developed pressure decrease rate (-dp/dt(max)) and heart rate (HR) were recorded at 0, 15, 30, 45 and 60 min of treatment. After the experiment, immunohistochemical examination was performed to detect the presence of C3a or C5b-9 in the myocardium of the isolated hearts.Results Compared the IPZ group, hearts treated with NPZ showed a slight depression on ST segments of epicardial ECG at 15 min, a significant elevation between 30min to 60min, a decrease in CF, CO, LVP(max), +dp/dt(max) and -dp/dt(max), and an increase in HR at 15 min. The observed alterations in CF, CO, LVP(max), +dp/dt(max) and -dp/dt(max) remained decreased, while the HR remained increased until the end of the protocol. The all above parameters of hearts treated with NPZC were similar to the control group (IPZ) at any given time. Immunohistochemical examination showed positive signals of C3a and C5b-9 in the myocardium of hearts treated with NPZ. C3a was positive in NPZC, and C3a and C5b-9 were negative in IPZ.Conclusions Activated human complements directly damage isolated guinea pig working hearts, and CD59 offers a significant protection against the injuries.     

无氧阈强度运动治疗对慢性缺血性心脏病患者运动耐力的影响

目的 探讨无氧阈强度运动治疗对血管重建后的慢性缺血性心脏病患者运动耐力的影响.方法 43例完全血管重建后的慢性缺血性心脏病患者(3例冠脉搭桥,22例陈旧性心肌梗死及18例不稳定性心绞痛),实施了心肺运动负荷试验(CPET)和3个月的无氧阈强度的运动治疗,其中32例患者完成了运动治疗; 3个月后复查CPET,比较患者运动耐力的前后变化.结果 无氧阈运动强度心率[(97±9)次/min]显著小于传统运动康复的最小运动强度心率[(112±7)次/min],小于运动试验的最初缺血强度心率[(115±11)次/min];3个月后运动处方执行组无氧阈氧耗量从(10.7±2.4 ) ml·min-1·kg-1到(12.6±2.9) ml·min-1·kg-1(P=0.04),有氧运动负荷能力从(37±18) J/s到(47±13) J/s(P=0.04),峰值氧耗量从(15.3±3.1) ml·min-1·kg-1到(20.6±4.2) ml·min-1·kg-1(P=0.02)及峰值运动负荷能力(68±12) J/s到(87±14) J/s(P=0.01),均明显增加;运动处方未执行组的无氧阈氧耗量从(11.0±2.7) ml·min-1·kg-1到(11.3±2.8) ml·min-1·kg-1,有氧运动负荷能力从(38±11) J/s到(37±9) J/s,峰值氧耗量从(15.3±2.9) ml·min-1·kg-1到(16.2±3.1) ml·min-1·kg-1] 和峰值运动负荷能力从(65±13) J/s到(73±16) J/s增加,差异均无统计学意义.结论 无氧阈强度显著小于慢性缺血性心脏病患者运动最初出现缺血时的强度;无氧阈强度运动治疗能有效地提高慢性缺血性心脏病患者的氧代谢水平和运动耐受力.

Abstract：

Objective To investigate the effects of exercise therapy at the intensity of anaerobic threshold (AT) for exercise tolerance in patients with chronic stable coronary artery disease. Methods Forty-three patients with chronic stable coronary artery disease (3 patients after coronary arterial bypass graft (CABG) surgery, 22 patients with old myocardial infarction and 18 unstable angina pectoris undergoing successful percutaneous coronary intervention (PCI) finished twice cardiopulmonary exercise test (CPET) and followed their rehabilitation program for 3 months. Thirty-two patients finished their aerobic exercise therapy based on their individual anaerobic thresholds while 11 patients had no exercise therapy. Results The heart rate at AT intensity (97±9 /min) was lower than their traditional minimal target heart rate (112±7 /min) and lower than heart rate (115±11 /min) at ischemic threshold post-CPET. The O2 consumption (10.7±2.4 to 12.6±2.9 ml·min-1·kg-1) （P=0.04） and workload (37±18 to 47±13 J/s) （P=0.04）at AT level and the O2 consumption (15.3±3.1 to 20.6±4.2 ml·min-1·kg-1,P=0.02) and workload（68±12 and 87±14 J/s, P=0.01）at peak levelmarkedly increased after 3 months in the exercise group. And the O2 consumption（15.3±2.9 to 16.2±3.1 ml·min-1·kg-1）and workload（65±13 to 73±16 J/s）at peak level mild increased after 3 months in the non-exercise group, but their O2 consumption （11.0±2.7 to 11.3±2.8 ml·min-1·kg-1）and workload （38±11 to 37±9 J/s）at AT level had no obvious change. Conclusion AT exercise intensity was lower than ischemic threshold post-CPET. Exercise therapy at the intensity of anaerobic threshold can improve oxygen capacity and exercise tolerance.     

白首乌提取物对血管性痴呆模型小鼠学习记忆功能的影响

目的 研究自首乌提取物对血管性痴呆模型小鼠学习记忆功能和抗氧化水平的影响.方法 小鼠反复脑缺血再灌注合并尾部放血降压建立痴呆模型;Morris水迷宫观察各组小鼠的学习记忆功能;快速断头处死,观测张口喘气时间;测定海马区及大脑皮质组织匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)水平.结果 与模型组相比,药物高剂量组逃避潜伏期缩短,差异具有显著性(P<0.01),正确探索时间延长(P<0.01);张口喘气时间延长[(19.3±1.6)s vs(15.7±0.7)s,P<0.01];SOD活性升高[(121.0±14.2)U·mgprot-1 vs(101.5±9.4)U·mgprot-1,P<0.05];MDA含量降低[(5.12±1.09)nmol·mgprot-1和(6.77±0.96)nmol·mgprot-1,P<0.01].结论 白首乌提取物对小鼠脑缺血再灌注损伤有保护作用,其机制可能与抗自由基损伤有关.     

参附注射液对糖尿病大鼠缺血再灌注损伤心肌DJ-1表达的影响

目的：观察参附注射液(SFI)对糖尿病大鼠缺血再灌注损伤(IR)心肌DJ-1表达的影响,并探究其对糖尿病IR心肌保护作用的分子机制。方法健康雄性SD大鼠腹腔注射60 mg/kg链尿佐菌素制备糖尿病模型。取糖尿病造模成功大鼠30只,随机数字表法分为三组(n=10)：假手术组(S组)、IR组、IR+SIF组。心肌IR模型采用结扎冠脉左前降支(LAD)30 min后松开120 min制备,S组只穿线包绕LDA而不结扎。IR+SFI组开胸前SFI以10 mL·kg-1·h-1持续泵注,开胸后以3 mL·kg-1·h-1持续输注至手术结束,其余两组泵注等量晶体液。检测SD大鼠心梗面积、血清肌酸激酶-MB(CK-MB)含量、心肌DJ-1表达,超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与S组比较,IR组大鼠的心梗面积[(49.0±5.3)%vs (0.0±0.0)%],CK-MB [(1982±275) U/L vs (1076±262) U/L]、MDA [(13.1±1.9) nmol/mg vs (7.6±1.1) nmol/mg]含量显著增加,差异均有统计学意义(P<0.05),而DJ-1表达[(0.65±0.14) vs (1.0±0.0)]和SOD活性[(79.0±19.3) U/mg vs (143±15.4) U/mg]显著降低,差异均具有统计学意义(P<0.05);与IR组相比,IR+SFI组大鼠的心梗面积[(35.7±5.3)%vs (49.0±5.3)%],CK-MB [(1364±228) U/L vs (1982±275) U/L]和MDA [(7.3±1.25) nmol/mg vs (13.1±1.9) nmol/mg]含量显著降低,差异均有统计学意义(P<0.05),而DJ-1表达[(0.9±0.14) vs (0.65±0.14)]和SOD活性[(119.4±14.6) U/mg vs (79.0±19.3) U/mg]显著降低,差异均有统计学意义(P<0.05)。结论 SFI能显著降低糖尿病心肌IR损伤的易损性,其机制可能与其上调心肌DJ-1表达、增强内源性抗氧化应激有关。     

urantide对小鼠急性肝细胞凋亡的影响及其机制

目的 探讨urotensinⅡ(UⅡ)特异性受体(UT)特异性拮抗剂urantide对急性肝细胞凋亡的影响及其机制.方法 雄性BALB/c小鼠按随机排列表法随机分为4组(每组6只):健康对照组、预处理对照组、模型组和预处理模型组.预处理对照组和预处理模型组给予尾静脉注射0.6 mg/kg urantide预处理,健康对照组和模型组则给予相同体积生理盐水.30min后模型组及预处理模型组立即以脂多糖联合D-半乳糖胺( D-GalN)腹腔注射诱导急性肝细胞凋亡小鼠模型,健康对照组和预处理对照组则给予相应体积的生理盐水.用原位末端转移酶标记(TUNEL)技术分析和半胱氨酸蛋白酶(caspase)-3活性测定检测肝细胞凋亡程度;用反转录(RT)-PCR及酶联免疫吸附试验( ELISA)方法检测肝组织及血浆前炎细胞因子肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ和白细胞介素(IL)-1β的表达与分泌水平.结果 模型组可见大量肝细胞凋亡,而预处理模型组肝细胞凋亡指数和caspase-3活性均明显低于模型组[(26±11)％比(77±20)％,(2.50±0.83) pmol·min-1·mg-1比(3.76±0.42) pmol·min -·mg-1,均P＜0.01];肝组织前炎细胞因子TNF-α、IL-1β和IFN-γ的mRNA相对表达水平及蛋白分泌水平也均明显低于模型组[1.69±0.47比3.57±0.79、0.31±0.02比0.46±0.06、2.81±0.72比3.35±0.84,(233±36) pg/ml比(441±157) pg/ml、(228 ±21) pg/ml比(364±20) pg/ml、(93.8±5.2) pg/ml比(180.3±4.3) pg/ml,均P＜0.01].结论 urantide可通过抑制前炎细胞因子的表达与分泌抑制脂多糖/D-GalN诱导的急性肝细胞凋亡.这表明UⅡ/UT受体系统在急性肝衰竭免疫炎性损伤中起关键性的作用,并有可能成为ALF药物治疗的新靶点.     

Trichloroethylene改变融合期内皮细胞一氧化氮和氧自由基的平衡

[目的] Trichloroethylene (TRI) 是一种与先天性心脏病发生有关的环境污染物,我们最近报道在分裂期的内皮细胞( EC)中, TRI可以通过减少热休克蛋白 90( hsp90)与内皮一氧化氮合酶( eNOS)结合,从而抑制刺激状态下一氧化氮( NO)产生和增加刺激状态下 eNOS依赖的超氧阴离子自由基(O2)产生和抑制血管内皮生长因子刺激的内皮细胞分裂.但 TRI在融合期 EC中的影响仍未清楚.本实验目的是探讨 TRI对融合期 EC的影响.[方法] TRI 5 μ mol/L 预处理融合期 EC,用钙离子导体 A23187 5 μ mol/L 刺激,然后,检测 NO与(O2)的产生以了解 EC和 eNOS的功能情况.用免疫印迹方法检查 eNOS,位点于丝氨酸 1179的磷酸化 eNOS,与 eNOS结合的 hsp90水平来决定 eNOS的激活状态.[结果] TRI减少 A23187刺激状态下亚硝酸盐和硝酸盐的产生从 (1.16 ± 0.15) nmol· mg- 1到 (0.91 ± 0.3) nmol· mg- 1 (P < 0.05);在对照情况下, L- NAME 增加 A23187 刺激状态下的(O2)产生从 (0.015 ± 0.007) nmol/(min· mg)到 (0.044 ± 0.008) nmol/(min · mg) (P< 0.05);在 TRI处理的培养中, L- NAME 减少 A23187刺激状态下的产生从 (0.057 ± 0.022) nmol/(min · mg)到 (0.039 ± 0.005) nmol/(min · mg) (P< 0.05),并且 oxypurinol减少 A23187刺激状态下的(O2)产生从 (0.057 ± 0.022) nmol/(min · mg)到 (0.034 ± 0.012) nmol/(min · mg) (P< 0.05); TRI 对 eNOS, p- eNOS 和 hsp90的表达无影响,但减少 hsp90与 eNOS的结合.[结论] 这些数据表明,在融合期 EC中, TRI 不但改变 hsp90与 eNOS 结合,使 eNOS 从产生 NO变为产生 (O2),而且激活第二氧化酶-黄嘌呤氧化酶,而增加(O2)产生.这些内皮功能的改变可能导致许多心血管疾病的发生.     

丹参多酚酸对大鼠缺血心肌线粒体酶的影响

目的:探讨丹参多酚酸对大鼠心肌缺血再灌注过程中线粒体Na+,K+-ATP酶和Ca2+-ATP酶的影响。方法:采用健康Wistar大鼠(n=40)建立Langendorff离体心脏灌注模型,并分为4组。对照组用Krebs液持续灌注120 min;缺血再灌注组平衡30 min后全心缺血30 min,再灌注60 min;丹参多酚酸预处理组平衡5 min后加入丹参多酚酸使其在灌流液中终浓度为14μmol/L,持续灌注25 min后全心缺血30 min,再灌注60 min;丹参多酚酸后处理组平衡30 min后全心缺血30 min,加入丹参多酚酸使其在灌流液中终浓度为14μmol/L,再灌注60 min。TTC染色观察心肌梗死面积,并用心肌梗死面积、乳酸脱氢酶(LDH)活性和心脏血流动力学(包括心率、主动脉流量、冠状动脉流量和心输出量)的变化评估大鼠心肌的损伤程度。利用差速离心法提取线粒体,并通过紫外分光光度法测定心肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶的活性。结果:丹参多酚酸预处理组的心肌梗死面积和LDH活性均显著低于缺血再灌注组([24.47±2.14)%比(34.30±2.31)%、(81.46±13.39)U/g比(142.6±6.46)U/g,均P<0.05],缺血再灌注组与后处理组无明显差异。丹参多酚酸对缺血再灌注心肌的血液动力学没有明显作用,只是冠脉流量有较小幅度的增加。丹参多酚酸预处理组线粒体Na+,K+-ATP酶和Ca2+-ATP酶活性显著高于缺血再灌注组([6.12±0.42)U/mg比(4.29±0.28)U/mg、(3.42±0.16)U/mg比(2.62±0.96)U/mg,均P<0.05],而与对照组无明显差异。丹参多酚酸后处理组线粒体Na+,K+-ATP酶和Ca2+-ATP酶活性与缺血再灌注组无统计学差异。结论:丹参多酚酸可以维持心肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶的活性,通过降低缺血再灌注对心肌线粒体的损伤保护心肌。     

慢性间歇低氧训练对大鼠心肌线粒体ATP酶的影响

目的: 研究慢性间歇低氧训练对大鼠心肌线粒体ATP酶含量及Na+、K+-ATP酶和Ca2+、Mg2+-ATP酶的影响。方法: 32只雄性Wistar大鼠按低氧训练方案分为慢性组11只、急性组12只、对照组9只。慢性组行慢性间歇低氧训练, 首先置于低压氧舱, 模拟海拔3 km, 每日持续4 h, 训练2周;再模拟海拔5 km, 每日持续4 h, 训练2周;最后模拟海拔8 km, 放置4 h。急性组立即置于模拟海拔8 km, 放置4 h。对照组则不行低氧训练。低氧期满后, 断头处死大鼠, 分离心肌线粒体, 测定ATP酶活性。结果: 慢性组ATP酶含量(9.04±4.71)μmol·mg-1·h-1, 均较急性组(4.96±1.17)μmol·mg-1·h-1和对照组(4.38±0.95)μmol·mg-1·h-1明显升高(P< 0.01)。慢性组Na+、K+-ATP酶活性(2.55±1.41)μmol·mg-1·h-1, 与急性组(2.66±1.07)μmol·mg-1·h-1和对照组(3.08±1.37)μmol·mg-1·h-1差异均无统计学意义(P>0.05)。慢性组Ca2+、Mg2+-ATP酶活性(1.17±0.34)μmol·mg-11·h-1与对照组(1.28±0.42)μmol·mg-1·h-1差异无统计学意义(P>0.05), 但较急性组(0.58±0.14)μmol·mg-1·h-1明显升高(P< 0.01)。结论: 慢性间歇低氧训练可增强Ca2+、Mg2+-ATP酶活性, 同时能够显著增加ATP酶含量, 有助于提高心肌运动功能, 使大鼠适应低氧环境。     

CSI监测在肾移植手术麻醉中的应用

目的 探讨麻醉深度指数(CSI)监测肾移植手术麻醉的效果.方法 选择我院2013年7月至2015年7月急诊拟行肾移植手术麻醉的患者120例,按照随机数字表分为N组(对照组)与C组(接受麻醉深度监测),两组患者均采用相同的药物进行麻醉诱导与维持.N组患者术中维持用药仅采用丙泊酚与瑞芬太尼.瑞芬太尼浓度为0.1~0.3μg·kg-1·min-1;丙泊酚效应室浓度设定为2~3μg/ml,术中丙泊酚的输注速度依靠靶控输注技术(TCI)的原理,由Graseby3500输注泵自动调节术中丙泊酚输注的速度与剂量.C组患者术中维持药物与N组患者一致,瑞芬太尼浓度为0.1~0.3μg·kg-1·min-1.以CSI指数40~60为目标调节丙泊酚输注速度,若CSI指数低于40则降低丙泊酚输注速度与剂量,CSI指数高于60则加大丙泊酚输注速度.观察并记录两组患者年龄、体质量、手术时间、术中维持所使用丙泊酚与瑞芬太尼总量、麻醉苏醒时睁眼时间、拔管时间、恢复室停留时间.结果 与N组比较,C组性别、年龄、体质量和手术时间比较差异无统计学意义(P>0.05);C组术中维持所用丙泊酚总量低于N组[(124.4±27.6)mg vs(167.6±42.8)mg],差异有统计学意义(P<0.05);C组术中维持所用瑞芬太尼总量[(1911.8±197.5)μg vs(1876.1±213.4)μg]及顺式阿曲库胺总量[(20.9±2.7)mg vs(21.3±2.1)mg]与N组比较差异无统计学意义(P>0.05);C组麻醉苏醒时睁眼时间[(24.4±6.3)min]、拔管时间[(26.6±8.9)min]及恢复室停留时间[(45.2±9.4)min]均低于N组[(42.5±8.2)min、(45.7±3.1)min、(72.4±12.6)min],差异有统计学意义(P<0.05).结论 CSI监测能合理指导麻醉药物用量及减少患者麻醉恢复时间.     

丙泊酚后处理对离体大鼠缺血再灌注心肌的保护作用及机制

目的研究丙泊酚后处理对离体大鼠缺血再灌注心肌细胞的保护作用以及对PI3K/Akt信号通路的影响。方法选用SD大鼠28只,按照随机数字表法分为缺血再灌注组(I/R组)、丙泊酚后处理组(PPC组)、丙泊酚后处理+Wortmannin后处理组(PPC+W组)、Wortmannin组(W组)。4组均建立Langendorff离体心肌缺血再灌注模型。各组心肌做相应分组处理后,观察缺血前及再灌注120 min时左室心功能变化情况,免疫组化检测Bcl-2蛋白的表达,Tunel法检测各组心肌细胞凋亡情况,Western blot测定p-Akt(Ser473)蛋白表达。结果与I/R组相比,PPC组LVEDP降低[(43.31±4.70)vs(29.93±3.72),P<0.05],+dp/dtmax和-dp/dtmax均明显升高[(1 140±138)vs(1 622±160),(749±99)vs(1 008±178),P<0.05],心肌组织Bcl-2蛋白和p-Akt的表达均增加[(0.171 9±0.012 1)vs(0.199 1±0.014 4),(0.241 4±0.053 9)vs(0.436 3±0.081 7),P<0.05],心肌细胞凋亡明显减少[(33.87±1.72)vs(29.84±1.83),P<0.05]。Wortmannin能阻断丙泊酚后处理的心肌保护效应(P<0.05)。结论丙泊酚后处理能减少离体大鼠缺血再灌注心肌细胞凋亡,发挥心肌保护作用,其保护机制与激活PI3K/Akt信号通路及上调Bcl-2蛋白的表达有关。     

Apelin-13对在体大鼠心肌缺血-再灌注损伤的影响及其信号转导途径

目的 研究心血管活性肽Apelin-13对在体大鼠心肌缺血-再灌注损伤的影响,并探讨其信号转导途径.方法 将雄性SD大鼠随机分为对照组(n=17)和Apelin-13组(n=15),建立在体大鼠心肌缺血-再灌注模型,对照组在再灌注前5 min给予生理盐水颈静脉注射,Apelin-13组给予0.1 mg/kg Apelin-13颈静脉注射,用伊文思蓝和TTC双重染色法测定心肌梗死区(IS)面积和缺血危险区(ARR)面积,计算IS面积占ARR面积的百分比(IS/ARR);TUNEL法测定心肌细胞凋亡;Western blotting法检测ERK1/2蛋白的表达.结果 Apelin-13组的IS/AAR和心肌细胞凋亡指数均显著低于对照组[(38.33±12.95)% vs (52.61±11.00)%,(0.21±0.02 vs 0.31±0.05](P<0.05);Apelin-13组p-ERK1/2的表达较对照组明显增加(1.15±0.16 vs 0.63±0.07)(P<0.05).结论 Apelin-13具有心肌保护作用,能够缩小心肌梗死面积,减少心肌细胞凋亡,其作用机制可能与ERK1/2MAPK激酶途径激活有关.