蕨麻石油醚部位的抗缺氧及其初步机制研究

 目的研究蕨麻石油醚部位的抗缺氧作用及初步机制。方法采用常压密闭缺氧模型、减压缺氧模型及异丙肾上腺素所致小鼠特异性心肌缺氧模型研究其抗缺氧作用;采用紫外分光光度法研究其对抗氧自由基对红细胞的氧化作用。结果蕨麻石油醚部位能明显延长常压密闭缺氧小鼠在密闭状态下的存活时间,降低整体耗氧量,并能显著增加减压缺氧状态下小鼠存活率。能明显对抗氧自由基对红细胞的氧化作用。结论蕨麻石油醚部位具有明显的抗缺氧作用,此作用可能与其抗自由基的能力有关。     

Neuroprotection of n-Butanol Extract from Roots of Potentilla anserina on Hypoxic Injury in Primary Hippocampal Neurons

Objective To investigate the protective effect of n-butanol extract from the roots of Potentilla anserina (NP) on hypoxic hippocampal neurons in neonatal rats. Methods Primary cultured hippocampal neurons were pretreated with different concentration of NP (0.25, 0.0625, and 0.0156 mg/mL) before incubation in a low oxygen (0.1%) environment for 4 h. Cell viability was evaluated by Trypan blue staining assay. Lactate dehydrogenase (LDH) released by neurons into the medium was measured. The activity of superoxide dismutase (SOD) in cell cytosol was determined using nitroblue tetrazolium. Morphological changes and mitochondrial function were observed by transmission electron microscopy. Results Hypoxic injury could decrease the cells viability of neuron, enhance LDH release (P < 0.05), decrease SOD activity, and increase mitochondrial injury. Pretreatment with NP significantly increased cell viability, decreased LDH release (P < 0.05), promoted SOD activity (P < 0.05), and remarkably improved cellular ultra-microstructure compared with the model group. Conclusion NP could protect the primary hippocampal neurons from hypoxic injury by attenuating mitochondrial cell death.     

黄芪总皂甙对病毒性心肌炎小鼠心肌损伤及肌浆网钙泵的影响

目的：观察黄芪总皂甙（ＡＳ）及黄芪注射液（ＡＩ）对病毒性心肌炎（ＶＭＣ）小鼠心肌损伤的保护作用及其对心肌肌浆网钙泵的影响。方法：腹腔接种柯萨奇Ｂ３亲心肌病毒株建立小鼠ＶＭＣ模型，分成模型组，ＡＳ组，ＡＩ组及正常对照组，分别观察各组小鼠死亡率，心肌病理变化，血清心肌肌钙蛋白Ｉ（ｃＴｎＩ）含量及心肌肌浆网钙泵（ＳＥＲＣＡ）活力等指标，结果：模型组死亡率高于对照组（Ｐ＝０．００４２）ＡＳ组及ＡＩ组（Ｐ〉     

Astragalosides rescue both cardiac function and sarcoplasmic reticulum Ca²⁺ transport in rats with chronic heart failure

The study investigated the beneficial effects of astragalosides (AS) on cardiac performance in rats with chronic heart failure. Chronic heart failure was produced by left anterior descending coronary artery ligation, and the therapeutic efficacy of astragalosides at 10, 20 and 40 mg/kg was evaluated. Five weeks after the operation, cardiac function was deficient and sarcoplasmic reticulum Ca²⁺‐ATPase (SERCA) activity was significantly reduced. Moreover, SERCA mRNA decreased, while expression of the SERCA down‐regulator phospholamban (PLB) was significantly increased. Phosphorylated phospholamban (P‐PLB), the form that does not inhibit SERCA, was also reduced by chronic heart failure. Treatment with AS improved left ventricle function and cardiac structure, reversed the depression of SERCA activity, and increased P‐PLB. These results suggest that the cardioprotective effect of AS may be due to the increase in P‐PLB protein, which disinhibits SERCA activity. Rescue of sarcoplasmic reticulum Ca²⁺ cycling by astragalosides could normalize excitation–contraction coupling and improve overall cardiac function. Copyright © 2011 John Wiley & Sons, Ltd.     

藏药蕨麻中多糖的含量测定研究

目的:测定藏药蕨麻中多糖含量,方法:采用苯酚-硫酸法测定.结果:蕨麻中多糖含量为１１.０８３%.     

扩心方对扩张型心肌病大鼠肌浆网Ca2+-ATP酶及受磷蛋白的影响

目的: 研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法: 50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组（每组各10只）,连续灌胃给药四周,另设10只正常对照。正常组和模型组以生理盐水灌胃。治疗结束后,对各组大鼠进行心脏超声检查,病理形态学检测,以及用反转录-聚合酶链反应和蛋白质免疫印迹法分别检测心肌组织中SERCA2α、PLB的mRNA和蛋白含量。结果: 与正常对照组相比, 模型组心功能减弱,出现心肌病理损害,SERCA2α与PLB表达异常;与模型组相比,扩心方组可以改善DCM大鼠的心功能、心肌病理学形态,同时,提高SERCA2αmRNA和蛋白表达、抑制PLB mRNA和蛋白表达。结论: 扩心能提高DCM大鼠的左心室射血分数和缩短率,减轻心肌损伤,改善心功能,同时调控DCM大鼠SERCA2α、PLB的表达。提示扩心方改善DCM大鼠心功能的作用机制,可能与纠正心肌肌浆网钙调控相关蛋白-SERCA2α、PLB的异常表达相关。     

Polysaccharide extracted from Potentilla anserina L ameliorate acute hypobaric hypoxia‐induced brain impairment in rats

High altitude cerebral edema (HACE) is a high altitude malady caused by acute hypobaric hypoxia (AHH), in which pathogenesis is associated with oxidative stress and inflammatory cytokines. Potentilla anserina L is mainly distributed in Tibetan Plateau, and its polysaccharide possesses many physiological and pharmacological properties. In the present study, the protective effect and potential treatment mechanism of Potentilla anserina L polysaccharide (PAP) in HACE were explored. First, we measured the brain water content and observed the pathological changes in brain tissues, furthermore, malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), and glutathione (GSH) were evaluated by kits. Finally, the protein contents and mRNA expressions of pro‐inflammatory (IL‐1β, IL‐6, TNF‐α, vascular endothelial cell growth factor [VEGF], NF‐κB, and hypoxia inducible factor‐1 α [HIF‐1α]) were detected by ELISA kits, RT‐PCR, and western blotting. The results demonstrated that PAP reduced the brain water content, alleviated brain tissue injury, reduce the levels of MDA and NO, and increased the activity of SOD and GSH level. In addition, PAP blocking the NF‐κB and HIF‐1α signaling pathway activation inhibited the generation of downstream pro‐inflammatory cytokines (IL‐1β, IL‐6, TNF‐α, and VEGF). Therefore, PAP has a potential to treat and prevent of HACE by suppression of oxidative stress and inflammatory response.     

The source of ca2+ for the spasmolytic actions of longicaudatine,a bisindole alkaloid isolated from Strychnos trinervis (Vell.) Mart. (Loganiaceae)

Longicaudatine, a tertiary bisindole alkaloid isolated from the root bark of Strychnos trinervis (Vell.) Mart. (Loganiaceae), antagonized in a noncompetitive manner, carbachol and histamine induced contractions of the guinea-pig ileum and bradykinin responses in the rat uterus. The respective pD₂' values (mean ± SE) were 4.61 ± 0.21, 4.98 ± 0.04 and 4.49 ± 0.01. Longicaudatine, unlike verapamil, had no effect on voltage dependent Ca²⁺ channels, as it failed to inhibit KCI or CaCl₂ induced contractions of guinea-pig ileum and depolarized rat uterus respectively. When compared with sodium nitroprusside, an antagonist of receptor operated Ca²⁺ channels, longicaudatine produced a slower and weaker inhibition of noradrenaline induced sustained contractions of rabbit aortic strips. However, in the aorta, the alkaloid antagonized the intracellular calcium dependent transient contractions of noradrenaline and longicaudatine (IC₅₀, 5.01 × 10^?7 M) was approximately 133 times more potent that procaine (IC₅₀, 6.68 × 10^?5 M), a known inhibitor of the release of Ca²⁺ from intracellular stores. Longicaudatine may exert nonspecific spasmolytic effects by acting on intracellular Ca²⁺ stores, rather than on depolarization dependent or receptor operated Ca²⁺ channels.     

New Findings on the Effects of Tannic Acid: Inhibition of L‐Type Calcium Channels,Calcium Transient and Contractility in Rat Ventricular Myocytes

Tannic acid (TA) is a group of water‐soluble polyphenolic compounds that occur mainly in plant‐derived feeds, food grains and fruits. Many studies have explored its biomedical properties, such as anticancer, antibacterial, antimutagenic, antioxidant, antidiabetic, antiinflammatory and antihypertensive activities. However, the effects of TA on the L‐type Ca²⁺ current (I_Ca‐L) of cardiomyocytes remain undefined. The present study examined the effects of TA on I_Ca‐L using the whole‐cell patch‐clamp technique and on intracellular Ca²⁺ handling and cell contractility in rat ventricular myocytes with the aid of a video‐based edge detection system. Exposure to TA resulted in a concentration‐ and voltage‐dependent blockade of I_Ca‐L, with the half maximal inhibitory concentration of 1.69 μM and the maximal inhibitory effect of 46.15%. Moreover, TA significantly inhibited the amplitude of myocyte shortening and peak value of Ca²⁺ transient and increased the time to 10% of the peak. These findings provide new experimental evidence for the cellular mechanism of action of TA and may help to expand clinical treatments for cardiovascular disease. Copyright © 2016 John Wiley & Sons, Ltd.     

杜仲对SD大鼠闭合骨折早期血生化指标的影响

目的:观察杜仲对SD大鼠闭合骨折早期血生化指标的影响,探讨杜仲对骨折闭合早期sD大鼠断骨的愈合作用和机理.方法:168只大鼠双盲随机分为骨折1 d、2 d、3 d、5 d、7 d、14 d、21 d共7个时间点组,每时间点组分设杜仲组和生理盐水组各半,分别以杜仲液、生理盐水灌胃.通过主动脉取血,分别采用偶氮肿Ⅱ法、直接...     

藏药蕨麻中三萜类化合物的结构研究

目的对藏药蕨麻主要化学成分进行结构鉴定。方法采用大孔吸附树脂和硅胶柱色谱及HPLC制备色谱技术进行分离纯化,应用化学反应、热分析、光谱与电磁波谱综合解析,特别是一维和二维超导核磁共振技术,鉴定化合物的结构。结果从蕨麻首次分离纯化的3个化合物单体为五环三萜皂苷类成分,即2,α3,β19α-三羟基-齐墩果酸-28-O-β-D-葡萄糖苷(24-deoxy-sericos ide,Ⅰ)、2,α3,β19β-三羟基-乌苏酸-28-O-β-D-吡喃半乳糖苷(anseri-nos ide,Ⅱ)和2-羰基-3,β19α-二羟基-乌苏酸-28-O-β-D-葡萄糖苷(2-oxo-pom o lic ac id-βD-g lucopyranosy l ester,Ⅲ),其中化合物Ⅱ为新化合物(命名为蕨麻苷)。结论此3个化合物为首次从蕨麻中分离得到,其中蕨麻苷经鉴定为新化合物。     

手针与电针对急性运动大鼠骨骼肌肌浆网Ca~(2+)的影响

目的:研究手针和电针对急性游泳运动大鼠骨骼肌肌浆网Ca2+含量、Ca2+-ATP酶活性的影响,探讨针灸增强运动能力的作用机制。方法:60只SD雄性大鼠随机分为空白对照组、模型组、手针组、电针组。手针组和电针组在急性游泳运动前运用针刺和电针两种不同的方法,对"大椎"命门"环跳"足三里"穴进行20 min刺激。其中手针组运用捻转手法,每间隔3~5 min运针1次(频率约为120~140次/min),每次运针30~60 s;电针组在针刺后接BX型电针仪,电针参数为音频脉冲调制波,频率为500~800 Hz,电流为0.20~0.25 mA。采用比色法分别测定骨骼肌肌浆网Ca2+含量、Ca2+-ATP酶活性指标。结果:骨骼肌肌浆网Ca2+含量模型组明显低于空白对照组、手针组和电针组(P<0.01);肌浆网Ca2+-ATP酶活性模型组亦明显低于空白对照组、手针组和电针组(P<0.05,0.01)。手针组肌浆网Ca2+-ATP酶活性比空白对照组明显升高(P<0.05);手针组Ca2+含量及Ca2+-ATP酶活性与电针组比较,两组间差异无显著性意义(P>0.05)。结论:针刺可以提高大鼠肌浆网Ca2+-ATP酶活性,增强肌浆网对细胞内Ca2+浓度的调节作用,参与肌肉兴奋收缩偶联并维持肌细胞胞质Ca2+稳态,这是针灸增强运动能力的重要因素之一。     

络合Ca~(2+)对山羊膀胱经经穴氧分压的影响

目的:探讨Ca2+和组织氧分压与经络现象之间的联系。方法:波尔杂交母山羊10只,用盐酸氯丙嗪(0.85 mg/kg)镇静后,采用乙二胺四乙酸二钠(EDTA-Na2200μL,0.05 kg/mol)络合穴区局部组织Ca2+,用新型组织氧分压传感针检测山羊膀胱经经穴"肝俞"大肠俞"关元俞"及旁开非经穴处氧分压的变化趋势。结果:1)膀胱经经穴的Ca2+电位明显高于旁开对照点(P<0.05,0.01);2)膀胱经经穴的组织氧分压高于旁开对照点(P<0.01);3)注射EDTA-Na2后,经穴的氧分压较注射前及注射生理盐水均明显升高(P<0.01),且经穴的氧分压高于旁开对照点(P<0.01)。结论:膀胱经穴Ca2+电位明显高于各自旁开对照点;注射EDTA-Na2络合Ca2+后氧分压明显升高。     

黄芩茎叶总黄酮对缺氧/复氧诱导的心肌细胞凋亡相关基因Bcl-2、Bax表达的影响

目的:探讨黄芩茎叶总黄酮(scutellaria baicalelensis stem leaf total flavonoid SSTF)对培养的乳鼠心肌细胞缺氧/复氧时凋亡相关基因Bcl-2、Bax蛋白表达的影响。方法:原代培养乳鼠心肌细胞,缺氧(hypoxia H)2h后,复氧(reoxygenation R)4h制备缺氧/复氧损伤模型。以免疫组织化学方法检测心肌细胞Bcl-2、Bax蛋白表达变化。心肌细胞损伤程度以心肌细胞存活率表示。结果:缺氧2h再复氧4h,心肌细胞Bcl-2蛋白阳性表达指数(positive expressionindex PEI)显著低于对照组(P<0.01),Bax蛋白PEI显著高于对照组(P<0.01)。细胞存活率明显低于对照组(P<0.05)。SSTF干预组Bcl-2蛋白PEI明显高于缺氧/复氧组(P<0.05),Bax蛋白PEI显著低于缺氧/复氧组(P<0.05),细胞存活率明显高于缺氧/复氧组(P<0.05)。结论:心肌细胞缺氧再复氧时,Bcl-2蛋白表达降低,Bax蛋白表达增强;细胞存活率降低。SSTF可通过上调Bcl-2蛋白表达,抑制Bax蛋白表达,减少细胞凋亡,从而减轻心肌细胞缺氧/复氧损伤。     

红景天对实验性心肌损伤大鼠心功能及肌浆网钙泵基因表达的影响

 目的探讨红景天对多柔比星所致心衰大鼠心功能及肌浆网钙泵(Ca²⁺-ATPase,SERCA2a)表达的影响。方法30只大鼠分为正常对照组(A组),多柔比星组(B组),红景天组(C组),采用多柔比星腹腔注射的方法建立心衰模型,给予红景天水提液腹腔注射保护,对照观察血流动力学的变化,采用半定量RT-PCR检测心肌钙泵mRNA的表达水平的变化。结果与A组相比,B组大鼠左室收缩压(LVSP)、左室压力上升下降最大速率(±dp/dt_max)均显著性降低(P<0.01),左室舒张末压(LVEDP)显著性增高(P<0.01),心肌肌浆网钙泵的mRNA表达水平显著性降低(P<0.05)。而C组,LVSP,±dp/dt_max显著性高于B组(P<0.05),但仍与A组有差异(P<0.05),LVEDP较B组显著性降低(P<0.01),接近于A组(P>0.05),肌浆网钙泵的mRNA表达水平也较B组显著性增高(P<0.05)。结论红景天对多柔比星所致心衰大鼠的心功能有明显改善作用,其作用可能与红景天提高心肌钙泵基因表达水平有关。     

川芎嗪预处理对体外培养乳鼠心肌细胞低氧再给氧损伤的保护作用

用体外培养乳鼠心肌细胞观察了川芎嗪预处理对低氧／再给氧损伤的保护作用。结果表明：川芎嗪20μg/ml能明显提高低氧3小时／再给氧0、30、60、120分钟时细胞搏动和细胞生存率,使乳酸脱氢酶（LDH）、丙二醇（MDA）释放减少,超氧化物歧化酶（SOD）释放增多。这种保护作用与低氧预处理（HP）产生的结果一致。上述结果表明川芎嗪预处理对体外培养乳鼠心肌细胞低氧／再给氧所致损伤具有保护作用,其机制可能与提高心肌细胞抗氧化酶活性,清除氧自由基有关。     

薯蓣皂苷对缺氧/复氧心肌细胞损伤的抗氧化作用研究

目的评价薯蓣皂苷对心肌细胞的抗氧化保护作用。方法对培养的乳鼠心肌细胞造成缺氧/复氧损伤模型,比较正常对照组、缺氧/复氧损伤组、以及薯蓣皂苷各组(10ml/L、30ml/L、100ml/L)培养液中超氧化物歧化酶(SOD)、丙二醛(MDA)和一氧化氮(NO)含量。结果缺氧/复氧损伤组SOD活性明显降低,MDA和NO含量较正常显著升高;而薯蓣皂苷中、高剂量组上述指标都有不同程度的改善。结论薯蓣皂苷对缺氧/复氧造成的心肌细胞损伤具有保护作用,并呈现剂量依赖关系,作用机制与增强细胞抗氧化作用、减轻自由基和脂质过氧化物导致的细胞膜损伤有关。     

Bisnordihydrotoxiferine and vellosimine from Strychnos divaricans root: Spasmolytic properties of bisnordihydrotoxiferine

Bisnordihydrotoxiferine and vellosimine, two tertiary indole alkaloids have been isolated from the root of Strychnos divaricans. Bisnordihydrotoxiferine antagonized in a nonspecific manner, oxytocin and acetylcholine induced contractions in the rat uterus and acetylcholine and histamine responses in the guinea-pig ileum. Bisnordihydrotoxiferine, like verapamil, produced effects on voltage dependent Ca²⁺ channels. For example in guinea-pig ileum, bisnordihydrotoxiferine (pD'₂ 3.92±0.09) and verapamil (pD'₂ 6.00±0.11) inhibited KCl induced contractions. Furthermore, bisnordihydrotoxiferine (pD'₂ 4.37±0.02) and verapamil (pD'₂ 6.83±0.10) also antagonized CaCl₂ induced contractions of K⁺-depolarized rat uterus. When compared with sodium nitroprusside, an antagonist of receptor operated Ca²⁺ channels, bisnordihydrotoxiferine had no effect. However, in the aorta, the alkaloid (IC₅₀, 6.10 × 10^?6M) antagonized the intracellular calcium dependent transient contractions of noradrenaline and it was about four times more potent than procaine (IC₅₀, 2.30 × 10^?5M), a known inhibitor of the release of Ca²⁺ from intracellular stores. Bisnordihydrotoxiferine may produce nonspecific spasmolytic actions mainly by inhibiting intracellular calcium mobilization and to a lesser extent by inhibiting voltage dependent calcium channels in smooth muscles.     

诱导HEK293细胞增殖中钙离子及TRPC6的变化及意义

目的建立AngⅡ诱导人胚肾细胞增殖模型,探讨增殖效果与Ca2+变化及TRPC6表达的关系。方法用不同浓度AngⅡ诱导细胞,在不同时间(0,6,12,24,48 h),予LOS干预24 h后用激光扫描共聚焦显微镜测Ca2+变化,RT-PCR、Western Blot检测TRPC6表达情况。采用MTT法及台盼蓝法检测细胞增殖及活性,确定最适浓度及时间构建增殖模型。结果 AngⅡ10-6mol/L诱导HEK293 24 h后细胞增殖及Ca2+升高明显,LOS干预后细胞增殖及Ca2+明显下降。TRPC6表达(+/-),胞膜、胞质定位不清。结论 HEK293细胞自身无明显表达TRPC6,可利用此表达系统进一步研究外源性TRPC6与Ca2+变化的关系。AngⅡ诱导HEK293细胞增殖与Ca2+升高有关,具有一定的时间和剂量依赖性。     

栀子苷预处理对H9C2心肌细胞缺氧/复氧损伤的影响及机制研究

[目的]探讨栀子苷预处理对H9C2心肌细胞缺氧/复氧损伤的影响及机制。[方法]体外培养H9C2心肌细胞,制备缺氧/复氧损伤模型。实验分为正常对照组,缺氧/复氧模型组,栀子苷预处理组。分光光度法测定细胞培养液中肌酸激酶(CK)、乳酸脱氢酶(LDH)、丙二醛(MDA)含量以及过氧化物歧化酶(SOD)活性,四甲基偶氮唑蓝(MTT)法检测细胞存活率,Annexin V-FITC/PI双染法流式细胞术测定细胞凋亡率。[结果]与模型组比较,栀子苷预处理组能明显减少心肌细胞CK、LDH的漏出量,提高SOD活力,降低MDA含量,同时提高心肌细胞的存活率,减少心肌细胞的凋亡。[结论]栀子苷预处理对H9C2心肌细胞的缺氧/复氧损伤具有保护作用,其机制可能与增强心肌抗氧化能力、清除氧自由基以及抑制心肌细胞凋亡有关。