PKC抑制剂对力竭运动大鼠肾脏裂孔膜蛋白表达的影响*

目的: 观察大鼠在一次性力竭运动后肾脏裂孔膜蛋白的表达水平,探究PKC抑制剂对其蛋白表达水平的影响,揭示PKC在运动性蛋白尿形成中的作用机制。方法: SD雄性大鼠30只随机分为对照组(C)、运动组(E)、运动联合PKC抑制剂组(EPI),每组10只。E组和EPI组大鼠分别进行一次性跑台力竭运动(25 m/min),EPI组大鼠运动前1 d及1 h腹腔注射PKC抑制剂白屈菜红碱(chelerythrine,5 mg/kg),C组和E组注射相应体积的生理盐水。运动后即刻麻醉后,取血液、尿液及肾脏组织,使用化学比色法检测尿蛋白、尿酸、尿糖、血尿素、血尿酸、血糖水平,使用荧光探针法检测肾脏ROS水平,使用Western blot法检测肾脏PKC、Nox2、Nox4、nephrin、podocin蛋白表达。结果: ①与C组相比,E组尿蛋白、尿酸、尿糖、血尿素、血尿酸显著增多(P＜0.05),血糖显著减少(P＜0.01),肾脏ROS生成显著增多(P＜0.01),肾脏nephrin、podocin蛋白表达明显降低(P＜0.05),PKC、Nox2、Nox4蛋白表达明显增多(P＜0.05);②与E组比,EPI组尿蛋白、尿糖、血尿素显著减少(P＜0.05),血糖显著增加(P＜ 0.01),肾脏ROS生成显著降低(P＜0.01),EPI组肾组织中nephrin、podocin蛋白表达明显增加(P＜0.05),PKC、Nox2蛋白表达明显降低(P＜0.05)。结论: 一次性力竭运动通过PKC/NOX/ROS途径使大鼠肾脏裂孔膜蛋白nephrin、podocin表达下调;PKC抑制剂缓解力竭运动导致的肾脏裂孔膜蛋白表达下降,预防运动性蛋白尿的发生。     

杜仲多糖改善运动疲劳的研究

目的：评价杜仲多糖（EUP）抗运动性疲劳的效果。方法：40只昆明小鼠建立一个5周的小鼠游泳模型,将小鼠分为运动对照组、蔗糖对照组、高剂量EUP组和低剂量EUP组（n=10）,服药采取灌胃的方式。训练期结束后,测定各组小鼠的体重变化与游泳力竭时间、力竭游泳后动物的血糖浓度、血乳酸（BLA）浓度、血尿素氮（BUN）浓度、肌酸激酶（CK）活性、肝糖原、肌糖原含量变化。结果：EUP高剂量与低剂量组小鼠较对照组体重增加明显,E UP高剂量游泳力竭时间显著延长（P<0.05）、血清CK活性和BUN含量显著下降（P <0.05）,但血糖、肝肌糖原以及BLA的水平变化不明显。结论：EUP具有抗运动性疲劳的作用,其机理与其调节机体糖代谢、节约蛋白质有关。     

力竭运动后不同时相大鼠心电图、心功能变化及Nrf2的作用

目的：研究大鼠力竭运动及运动结束后心电图、心功能的动态变化规律及转录因子E2相关因子（Nrf2）相关的氧化应激变化,为运动性心脏损伤防治提供依据。方法：SD大鼠随机分为5组（n=6）：对照组（Con）组、力竭组（EE）、力竭恢复6 h,12 h,24 h组（EER6、EER12、EER24组）。急性力竭游泳建立损伤模型。分别对各组动物进行心电图描记,压力容积导管检测心功能改变,ELISA法观测血清ROS,Nrf2,GPX及CAT变化。结果：① EE组心率（HR）,收缩末期压力（Pes）,发展压,动脉弹性,压力上升,下降最大速率（dP/dt_max、-dP/dt_min）降至最低。舒张末期压力容积、收缩末期容积、搏出量、Tau值增大。EER6、EER12、EER24组HR、Pes、dP/dt_max、-dP/dt_min与EE组相比均差异显著。②EE组、EER6、EER12、EER24组与Con组相比心率加快,QT间期延长,P波R波ST段数值增高,但恢复各组与EE组相比无统计学意义。③EE组大鼠血清ROS、Nrf2含量升高,GPX含量降低,CAT在EER6组降至最低。④血清Nrf2水平与ROS,-dP/dt_min呈正相关,与HR、Ea呈负相关。血清ROS水平与EF,-dP/dt_min呈正相关,与HR、Ea、dP/dt_max呈负相关。结论：力竭运动后心脏生物电改变,舒缩功能均受损,以舒张功能减退突出,随力竭恢复时间延长,心脏舒缩功能逐步恢复,这与Nrf2调节GPX,CAT降低氧化应激有关。     

一次性力竭运动致大鼠骨骼肌氧化应激的机制

目的:观察一次性力竭运动对大鼠骨骼肌氧化应激相关酶表达的影响。方法:雄性SD大鼠40只,分为4组(n=10),分别为对照组(C组)、力竭运动组(E组)、运动+PKC抑制剂组(EC组)、运动+NOX抑制剂组(EA组)。三组运动大鼠进行3 d的跑台适应性运动(5 m/min,1次/日,无坡度),然后休息1 d;EC组于运动前1 d和运动前1 h注射PKC抑制剂白屈菜红碱(5 mg/kg),EA组同期注射NADPH氧化酶抑制剂Apocynin(10 mg/kg),C组和E组注射同等剂量生理盐水;三组运动大鼠进行一次性跑台力竭运动,力竭后取大鼠的跖肌,DCF荧光探针检测活性氧(ROS),Western blot分析NOX2、NOX4、3-NT,免疫沉淀分析PKC、NOX2、NOX4。结果:与C组相比,E组的ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著增加(P＜0.01,P＜ 0.05),EC组、EA组ROS无显著差异(P＞0.05),EC组NOX4蛋白表达显著增加(P＜0.05);与E组相比,EC组和EA组的ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著降低(P＜ 0.01,P＜0.05)。结论:力竭运动诱导骨骼肌NOX2、NOX4蛋白表达增加,PKC通过调控NOX2介导ROS的生成。     

筋骨草提取物抗运动疲劳的作用

目的：探讨筋骨草的抗运动性疲劳作用。方法：将120只雄性昆明种小鼠随机平均分成安静组、运动组、阳性对照组和筋骨草低、中、高剂量组（n=10）。其中低、中、高剂量组小鼠分别按100 mg/kg、200 mg/kg、400 mg/kg体重连续灌胃筋骨草提取物30 d,阳性对照组小鼠按200 mg/kg体重灌胃西洋参胶囊颗粒,安静组和运动组小鼠以等体积生理盐水灌胃。动物试验结束后,分析各组小鼠运动力竭时间、血清生理生化指标（包括血乳酸、血尿素氮、血糖、总胆固醇、甘油三酯含量）、肝糖原与肌糖原含量,以及股四头肌、肝脏和心脏组织的抗氧化指标（包括谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶和丙二醛）。结果：中、高剂量组小鼠的运动力竭时间、红细胞数量、血红蛋白含量、血糖浓度、肝糖原与肌糖原含量,以及器官组织中谷胱甘肽过氧化物酶、超氧化物歧化酶和过氧化氢酶活力均明显高于运动对照组,而血清乳酸含量、血清尿素氮、血清甘油三酯与总胆固醇含量,以及器官组织中丙二醛含量明显低于运动对照组,中剂量的筋骨草提取物的作用效果优于同剂量的西洋参胶囊颗粒。结论：筋骨草通过提高机体的抗氧化功能而达到抗运动性疲劳作用。     

激活多巴胺I类受体对氧化性低密度脂蛋白诱导的人单核细胞THP-1分泌NO/NOS的影响

目的：探讨激活多巴胺Ⅰ类受体（DR1）对氧化型低密度脂蛋白（ox-LDL）诱导的人单核细胞（THP-1）分泌一氧化氮/一氧化氮合酶（NO/NOS）的影响及可能机制。方法：THP-1细胞经佛波酯PMA诱导分化,分为正常对照组（control）,氧化型低密度脂蛋白处理组（ox-LDL）,DR1激动剂干预组（SKF）,DR1阻断剂干预组（SCH）,ERK阻断剂干预组（PD98059）;应用油红O染色法鉴定泡沫细胞;硝酸还原法检测NO、NOS的变化情况;免疫荧光和Western blot检测各组细胞蛋白表达情况。结果：ox-LDL刺激48 h可形成泡沫细胞;DR1在THP1细胞上表达,ox-LDL刺激后,DR1蛋白表达降低（P<0.01）;激活DR1受体能够明显抑制由ox-LDL引起的NO、iNOS增多（P<0.01）;在MAPK阻断剂PD98059存在的情况下,SKF的作用部分丧失。结论：激活DR1受体可抑制ox-LDL引起的THP-1细胞NO的大量产生,此过程可能由ERK信号通路所介导。     

针刺对大鼠运动性骨骼肌损伤内质网应激的干预作用及机制

目的: 观察针刺对大鼠运动性骨骼肌损伤内质网功能酶SERCA、PDI、内质网应激标志蛋白GRP78和PERK通路的影响,探讨针刺防治运动性骨骼肌损伤的内质网途径作用机制。方法: 8周龄雄性SD大鼠随机分为空白对照组(C组,n=6)、单纯运动组(E组,n=30)、针刺对照组(A组,n=30)和运动针刺组(EA组,n=30)。其中,E组和EA组通过一次离心运动建立运动性骨骼肌损伤模型,EA组在运动后即刻于大鼠小腿跟腱上0.5 cm施以针刺干预,A组在同期施以针刺干预。各组根据运动和针刺干预后不同取材时间点分为0 h/12 h/24 h/48 h/72 h亚组(n=6),在对应时相取比目鱼肌进行指标测试。透射电镜观察肌纤维超微机构;ELISA法测定Ca²⁺-ATP酶(SERCA)和蛋白二硫键异构酶(PDI)含量;Western blot检测内质网应激标志蛋白GRP78及p-PERK、p-eIF2α表达。结果: 与C组比较,A组指标各时相均无显著差异(P＞0.05),E组肌纤维超微结构出现不同损伤,SERCA含量0 h至48 h均显著降低(P＜0.05),PDI含量0 h显著升高(P＜0.05),GRP78表达0 h至72 h均显著升高(P＜ 0.05),p-PERK表达0 h至24 h显著升高(P＜0.05), p-eIF2α表达与p-PERK一致;与E组对应时相比较,EA组肌纤维超微结构明显改善,SERCA含量48 h和72 h显著升高(P＜0.05),PDI含量0 h至72 h均显著升高(P＜0.05),GRP78表达0 h至72 h均显著降低(P＜0.05),p-PERK和p-eIF2α表达12 h和24 h显著降低(P＜0.05)。结论: 针刺可有效改善一次大负荷离心运动后导致的运动性骨骼肌损伤并缓解内质网应激,其机制可能与上调蛋白二硫键异构酶PDI以及抑制内质网应激PERK通路有关。     

运动对高脂饲料喂养大鼠骨骼肌和肝脏组织脂联素-脂联素受体-腺苷酸活化蛋白激酶通路的影响

目的：探讨运动影响高脂喂养大鼠骨骼肌和肝脏组织在腺苷酸活化蛋白激酶（AMPK）通路上的可能机制。方法：40只雄性SD大鼠随机分为4组（n=1 0）：正常对照组（C组）：正常饮食不运动;正常饮食运动组（E组）：正常饮食同时进行10周游泳运动;高脂饮食对照组（H组）：高脂饲料喂养不运动;高脂饮食运动组（HE组）：高脂饲料喂养同时进行10周游泳运动。采用实时荧光定量PCR检测大鼠骨骼肌和肝脏脂联系受体1（AdipoR1）,Adipo R2 mRNA表达,Western blot检测大鼠骨骼肌和肝脏腺苷酸活化蛋白激酶α（AMPKα）蛋白表达及磷酸化水平。结果：股四头肌AdipoR1 mRNA、肝脏AdipoR2 mRNA表达在H组显著低于C组（P<0.05）;HE组股四头肌和肝脏AMPKα（Thr172）磷酸化水平显著高于H组（P<0.05）,分别较H组高43.2%和51.1%。结论：高脂喂养导致大鼠骨骼肌和肝脏A dipoR1/R2 mRNA表达下调,运动提高了大鼠骨骼肌和肝脏AMPKα（Thr172）磷酸化水平。     

心肺运动指导个体化心衰患者康复的初步总结报告*

目的: 慢性心力衰竭患者（CHF）康复治疗和二级预防已经是临床共识,但用心肺运动试验（CPET）指导制定个体化运动处方国内尚少。方法: 选择10例CHF完成CPET评估,随机分为两组（n=5）：对照组进行没有运动;运动组增加△50%W功率运动30 min/d,每周5 d,共12周。在治疗前和3个月后分别进行评估。结果: 两组患者没有显著差异（P>0.05）。运动组12周后,运动持续时间从8 min显著提高到23 min（P<0.001）; 6分钟步行距离从394 m显著提高到470 m（P<0.05）;生活质量评分从25分显著降低至3分（P<0.01）。而对照组治疗前后均没有显著改变（P>0.05）;治疗前后的改变均显著小于运动组（P<0.01）。结论: 在CPET客观定量功能评估指导制定个体化高强度运动康复安全有效,值得大力推广应用。     

经皮冠状动脉腔内血管成形术改变稳定性冠心病患者整体功能的临床研究*

目的: 应用症状限制极限负荷心肺运动试验（CPET）评估稳定性冠心病患者经皮冠状动脉腔内血管成形术（PCI）治疗前后的整体心肺功能变化。方法: 入选2014年8月至12月在本院经冠脉造影和心脏超声等检查诊断为稳定性冠心病患者59例,择期行PCI治疗31例（PCI组）,另单纯药物保守治疗28例为对照组。患者治疗前、后均进行CPET。结果: 所有患者均安全完成CPET,无任何并发症。药物对照组治疗前后所有功能指标均无明显变化（P>0.05）。PCI组治疗后仅无氧阈、峰值摄氧量和峰值氧脉搏比治疗前明显提高（P<0.05）,其他指标变化不显著（P>0.05）。CPET评估个体化分析发现PCI组治疗后升高（≥10%）峰值摄氧量和峰值氧脉搏比例明显高于对照组（P<0.05）。结论: PCI通过冠状动脉血运重建可明显改善患者心肺功能,提高运动能力。CPET是客观定量评估冠心病治疗效果的一种客观、定量、安全、有效方法。     

Effect of apocynin treatment on renal expression of COX-2, NOS1, and renin in Wistar-Kyoto and spontaneously hypertensive rats

Macula densa (MD) cells of the juxtaglomerular apparatus (JGA) synthesize type 1 nitric oxide synthase (NOS1) and type 2 cyclooxygenase (COX-2). Both nitric oxide (NO) and prostaglandins have been considered to mediate or modulate the control of renin secretion. Reactive oxygen species (ROS) produced locally by NADPH oxidase may influence NO bioavailability. We have tested the hypothesis that in hypertension elevated ROS levels may modify the expression of NOS1 and COX-2 in the JGA, thereby interacting with juxtaglomerular signaling. To this end, spontaneously hypertensive rats (SHR) and Wistar-Kyoto control rats (WKY) received the specific NADPH oxidase inhibitor, apocynin, during 3 wk. Renal functional and histochemical parameters, plasma renin activity (PRA), and as a measure of ROS activity, urinary isoprostane excretion (IP) were evaluated. Compared with WKY, IP levels in untreated SHR were 2.2-fold increased, and NOS1 immunoreactiviy (IR) of JGA 1.5-fold increased, whereas COX-2 IR was reduced to 35%, renin IR to 51%, and PRA to 7%. Apocynin treatment reduced IP levels in SHR to 52%, NOS1 IR to 69%, and renin IR to 62% of untreated SHR, whereas renin mRNA, COX-2 IR, glomerular filtration rate, PRA, and systolic blood pressure remained unchanged. WKY revealed no changes under apocynin treatment. These data show that NADPH oxidase is an important contributor to elevated levels of ROS in hypertension. Upregulation of MD NOS1 in SHR may have the potential of blunting the functional impact of ROS at the level of bioavailable NO. Downregulated COX-2 and renin levels in SHR are apparently unrelated to oxidative stress, since apocynin treatment had no effect on these parameters.     

石菖蒲及α-细辛醚对疲劳运动大鼠学习记忆的影响及其机制*

目的: 研究石菖蒲及其活性成分-α-细辛醚改善疲劳运动大鼠学习记忆的作用及其机制。方法: 80只SD大鼠随机分为正常对照组(A)、单纯运动组(B)、运动+α-细辛醚低、中、高剂量组(C、D、E)、运动+石菖蒲低、中、高剂量治疗组(F、G、H),每组10只。并在疲劳运动开始前2 h分别以0.10、0.50 和 1.00 mg α-细辛醚,灌胃C、D、E组,以0.12、1.20和4.80 g. kg^-1. wt^-1石菖蒲提取物,灌胃 F、G、H组。实验结束后采用水迷宫实验进行学习记忆检测,采用生化法检测SOD、NOS活性和MDA含量,免疫印迹法检测海马nNOS蛋白表达水平。结果: 实验后E和H组大鼠逃避潜伏期、海马脑组织MDA含量低于B、C、D、F和G组;穿越平台次数、海马脑组织SOD和NOS活性、nNOS蛋白表达高于B、C、D、F和G组,P均＜0.01。A、E、H组大鼠海马脑组织SOD活性依次为A＞E＞H组, 而MDA含量则相反,P均＜0.01;E组大海马脑组织NOS活性和nNOS蛋白表达低于A和H组,P＜0.01或P＜0.05,但A和H组,差异无显著性,P均＞0.05。A、E、H组大鼠逃避潜伏期和穿越平台次数,差异均无显著性,P均＞0.05。结论: 石菖蒲及α-细辛醚改善疲劳运动大鼠学习记忆的作用基本一致,其机制与纠正海马自由基代谢失衡和上调海马nNOS/NO信号有关。     

Effect of NADPH oxidase inhibition on cardiopulmonary bypass-induced lung injury

Cardiopulmonary bypass (CPB) causes acute lung injury. Reactive oxygen species (ROS) from NADPH oxidase may contribute to this injury. To determine the role of NADPH oxidase, we pretreated pigs with structurally dissimilar NADPH oxidase inhibitors. Low-dose apocynin (4-hydroxy-3-methoxy-acetophenone; 200 mg/kg, n = 6), high-dose apocynin (400 mg/kg, n = 6), or diphenyleneiodonium (DPI; 8 mg/kg) was compared with diluent (n = 8). An additional group was treated with indomethacin (10 mg/kg, n = 3). CPB was performed for 2 h with deflated lungs, complete pulmonary artery occlusion, and bronchial artery ligation to maximize lung injury. Parameters of pulmonary function were evaluated for 25 min following CPB. Blood chemiluminescence indicated neutrophil ROS production. Electron paramagnetic resonance determined the effect of apocynin and DPI on in vitro pulmonary endothelial ROS production following hypoxia-reoxygenation. Both apocynin and DPI attenuated blood chemiluminescence and post-CPB hypoxemia. At 25 min post-CPB with Fi(O(2)) = 1, arterial Po(2) (Pa(o(2))) averaged 52 +/- 5, 162 +/- 54, 335 +/- 88, and 329 +/- 119 mmHg in control, low-dose apocynin, high-dose apocynin, and DPI-treated groups, respectively (P < 0.01). Indomethacin had no effect. Pa(O(2)) correlated with blood chemiluminescence measured after drug administration before CPB (R = -0.60, P < 0.005). Neither apocynin nor DPI prevented the increased tracheal pressure, plasma cytokine concentrations (tumor necrosis factor-alpha and IL-6), extravascular lung water, and pulmonary vascular protein permeability observed in control pigs. NADPH oxidase inhibition, but not xanthine oxidase inhibition, significantly blocked endothelial ROS generation following hypoxia-reoxygenation (P < 0.05). NADPH oxidase-derived ROS contribute to the severe hypoxemia but not to the increased cytokine generation and pulmonary vascular protein permeability, which occur following CPB.     

Apocynin attenuates oxidative stress and hypertension in young spontaneously hypertensive rats independent of ADMA/NO pathway

Both NADPH oxidase-derived reactive oxygen species (ROS) and asymmetric dimethylarginine (ADMA) are increased in hypertension. Apocynin, an NADPH oxidase inhibitor, could inhibit ROS, thus we tested whether apocynin can block NADPH oxidase and prevent increases of ADMA and blood pressure (BP) in spontaneously hypertensive rats (SHRs). SHRs and Wistar Kyoto (WKY) rats, aged 4 weeks, were assigned to four groups: untreated SHRs and WKY rats, SHRs and WKY rats that received 2.5 mM apocynin for 8 weeks. BP was significantly higher in SHRs compared to WKY rats, which was attenuated by apocynin. Apocynin prevented p47phox translocation in SHR kidneys, but not the increase of superoxide and H(2)O(2). Additionally, apocynin did not protect SHRs against increased ADMA. Apocynin blocks NADPH oxidase to attenuate hypertension, but has little effect on the ADMA/nitric oxide (NO) pathway in young SHRs. The reduction of ROS and the preservation of NO simultaneously might be a better approach to restoring ROS-NO balance to prevent hypertension.     

Role of the plasma membrane ROS-generating NADPH oxidase in CD34+ progenitor cells preservation by hypoxia

Hypoxia favored the preservation of progenitor characteristics of hematopoietic stem and progenitor cells (HSPCs) in bone marrow. This work aimed at studying the role of reactive oxygen species (ROS)-generating NADPH oxidase system regulated by hypoxia in ex vivo cultures of cord blood CD34⁺ cells. The results showed that NADPH oxidase activity and ROS generation were reduced in hypoxia with respect to normal oxygen tension. Meanwhile the ROS generation was found to be inhibited by diphenyleneiodonium (the NADPH oxidase inhibitor), or N-acetylcysteine (the ROS scavenger). Accordingly NADPH oxidase mRNA and p67 protein levels decreased in hypoxia. The analysis of progenitor characteristics, including the proportion of cultured cells expressing the HSPCs marker CD34⁺CD38⁻, colony production ability of the colony-forming cells (CFCs), and the re-expansion capability of the cultured CD34⁺ cells, showed that either 5% pO₂ or reduced ROS favored preserving the characteristics of CD34⁺ progenitors, and promoted the expansion of CD34⁺CD38⁻ cells as well. The above results demonstrated that hypoxia effectively maintained biological characteristics of CD34⁺ cells through keeping lower intracellular ROS levels by regulating NADPH oxidase.