胰升糖素样肽1对乳鼠心肌细胞缺氧/复氧损伤的保护机制

观察胰升糖素样肽1(glucagon-like peptide-1,GLP-1)对原代培养的心肌细胞在缺氧/复氧条件下损伤的影响及机制.结果显示,缺氧/复氧使乳酸脱氢酶活性[(210.0±11.5)对(101.4±6.5)U/L]、细胞凋亡率[(8.138±1.512)对(0.575±0.168)%]和半胱氨酸天冬氨酸蛋白酶3活性[(44.52±5.69)对(19.98±1.97),均P<0.01]均升高,GLP-1能直接作用心肌细胞,并对其在缺氧/复氧的损伤具有抑制作用[乳酸脱氢酶活性(190.2±9.0)U/L,细胞凋亡率(2.688±0.580)%,半胱氨酸天冬氨酸蛋白酶3活性30.34±4.18],该作用可能与磷脂酰肌醇3激酶(PBK)/Akt介导的抗细胞凋亡作用有关.     

丙泊酚对缺氧/复氧培养大鼠心肌细胞c-Fos表达及LDH的影响

目的 :研究丙泊酚对缺氧 /复氧培养大鼠心肌细胞 c- Fos表达和培养基乳酸脱氢酶 (L DH)含量的影响。方法 :建立 S D大鼠心肌细胞原代培养及缺氧 /复氧模型。将培养细胞按孔随机分为 6组 : 组为对照组 , 组为缺氧 /复氧组 ; 、 、 和 组为在缺氧前 10 min开始分别加入脂肪乳、丙泊酚 2 5μmol· L- 1 、5 0μmol· L- 1 和 10 0μmol· L- 1。用免疫组化法和灰度分析观察 3种剂量的丙泊酚对复氧 2 h c- Fos表达的影响。用生化比色法测定复氧3h时培养基中 L DH含量。结果 :缺氧 /复氧使心肌细胞 L DH漏出量明显增加 (P<0 .0 5 ) ,3种浓度的丙泊酚均能明显抑制细胞 L DH释放 (P<0 .0 5 )。 组与 组相比平均灰度值明显降低 (P<0 .0 5 ) ; 、 组与 组比较平均灰度值均显著增高 (P<0 .0 5 )。结论 :3种浓度的丙泊酚均有心肌保护作用且 5 0μmol· L- 1 和 10 0μmol· L- 1 浓度的丙泊酚能明显抑制缺氧 /复氧培养大鼠心肌细胞 c- Fos表达     

麦冬总皂甙对培养心肌细胞缺氧再给氧损伤的保护作用

目的:观察麦冬总皂甙(Ophiopogan Total Saponies,OTS)对培养心肌细胞缺氧再给氧损伤的保护作用。方法:以培养的心肌细胞为模型,在缺氧再给氧损伤细胞后,观察心肌细胞活力及搏动频率的变化,测定培养液上清乳酸脱氢酶的含量。结果:OTS 10 μg/L和40 μg/L可以提高损伤后心肌细胞的活力,提高心肌细胞的搏动频率,并使培养液上清乳酸脱氢酶的含量明显降低。结论:OTS对缺氧再给氧损伤的心肌细胞具有保护作用。     

地黄多糖对乳鼠心肌细胞缺氧/复氧损伤保护作用的研究

目的研究地黄多糖(RPS)对乳鼠心肌细胞缺氧/复氧损伤的保护作用。方法取出生3天的SD大鼠乳鼠分离心肌细胞,培养72h后分为6组:空白对照组、缺氧/复氧模型对照组、地黄多糖(10、20、40μg/ml)干预组和舒血宁注射液(SXN,100μg/ml)干预组,每组设10个复孔。各组细胞经药物干预6h后,通过倒置光学显微镜观察各组细胞形态变化,通过甲基四唑蓝(MTT)比色法测定细胞存活率,通过流式细胞术测定细胞凋亡率;测定各组细胞培养液中心肌酶[谷草转氨酶(AST)、磷酸肌酸激酶(CPK)、乳酸脱氢酶(LDH)]含量;测定各组细胞中抗氧化酶[超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)]活性和丙二醛(MDA)含量。结果与缺氧/复氧模型对照组相比,地黄多糖干预组心肌细胞形态明显好转,细胞存活率明显升高、凋亡率显著降低,培养液中AST、CPK、LDH含量显著降低,细胞中SOD、GSH-Px、CAT活性显著升高且MDA含量显著降低,其中以地黄多糖40μg/ml干预组效果最为显著,差异具有统计学意义(P<0.05,P<0.01)。结论地黄多糖能够有效改善缺氧/复氧损伤心肌细胞形态、提高细胞存活率、降低细胞凋亡率、改善抗氧化酶活性、抑制氧化应激损伤,提示地黄多糖对缺氧/复氧损伤心肌细胞具有保护作用。     

右美托咪定联合Tol样受体4抑制剂对缺氧复氧心肌细胞凋亡和炎症反应的影响及其机制

目的探讨右美托咪定(DEX)联合Toll样受体4(TLR4)抑制剂TAK-242对缺氧复氧(H/R)心肌细胞凋亡和炎症反应的影响及其机制。方法心肌细胞H9C2分为对照(Con)组、H/R组(H/R损伤)、DEX组(1.0μmol/L DEX,再行H/R损伤)、TAK-242组(30μmol/L TAK-242,再行H/R损伤)和DEX+TAK-242组(30μmol/L TAK-242及1.0μmol/L DEX,再行H/R损伤处理)。各组细胞复氧培养6 h后,采用MTT法、流式细胞仪、试剂盒、酶联免疫吸附法检测细胞增殖、凋亡、乳酸脱氢酶(LDH)释放率、白介素1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量,Western blot检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解的caspase-3(cleaved caspase-3)、TLR4和核因子B p65(NF-κB p65)蛋白表达。结果五组细胞凋亡率、LDH释放率、Bax、Bcl-2、cleaved caspase-3、TLR4、NF-κB p65蛋白表达水平、IL-1β、TNF-α含量比较差异均有统计学意义(F=316.938、330.004、839.933、169.750、378.365、476.535、298.527、99.219、293.498,P<0.05)。与Con组相比,H/R组细胞的凋亡率、LDH释放率、Bax、cleaved caspase-3、TLR4和NF-κB p65蛋白表达水平、细胞上清液中IL-1β和TNF-α含量均明显升高(均P<0.05),Bcl-2蛋白表达水平明显降低(均P<0.05)。与H/R组相比,DEX组、TAK-242组和DEX+TAK-242组的细胞凋亡率、LDH释放率、Bax蛋白、cleaved caspase-3、TLR4和NF-κB p65蛋白、IL-1β、TNF-α的表达水平均明显降低,Bcl-2蛋白表达水平明显升高(均P<0.05)。与DEX组、TAK-242组相比,DEX+TAK-242组的细胞凋亡率、LDH释放率、Bax、cleaved caspase-3、TLR4和NF-κB p65蛋白表达、IL-1β、TNF-α表达水平更低,Bcl-2蛋白的表达更高(均P<0.05)。结论DEX和TAK-242联合可协同抑制H/R引起的心肌细胞凋亡和炎症反应,其作用机制可能与协同抑制TLR4/NF-κB通路有关。     

Dose-response effects of milrinone on hemodynamics of newborn pigs with hypoxia-reoxygenation

OBJECTIVE: Neonatal asphyxia causes cardiogenic shock and pulmonary hypertension with decreased brain perfusion. We examined the dose-response of milrinone on systemic, pulmonary, and carotid circulations in a model of neonatal hypoxia-reoxygenation. DESIGN AND SETTING: Controlled, block-randomized study in a university research laboratory. SUBJECTS: Mixed breed piglets (1-3 days, 1.5-2.3 kg). INTERVENTIONS: In acutely instrumented piglets normocapnic alveolar hypoxia (10-15% oxygen) was induced for 2 h followed by reoxygenation with 100% oxygen (1 h) then 21% oxygen (3 h). At 2 h of reoxygenation after a volume loading (10 ml/kg) either saline or milrinone (bolus and infusion at 0.25, 0.5, or 0.75 microg/kg per minute) was given for 2 h in a blinded-randomized fashion (n = 7/group). MEASUREMENTS AND RESULTS: All milrinone-treated groups had higher cardiac output and stroke volume than those of saline-treated hypoxic controls, which showed progressive decline in these measurements. At 2 h of infusion plasma milrinone levels were significantly correlated with cardiac output (r = 0.6), which increased from pretreatment value in the group receiving 0.75 microg/kg per minute. Milrinone maintained mean arterial pressure; heart rate and pulmonary arterial pressure did not differ between groups. Milrinone prevented continued increases in systemic and pulmonary vascular resistances after hypoxia-reoxygenation. Milrinone infusion at higher doses increased common carotid flow. Milrinone-treated piglets had increased systemic and carotid oxygen delivery, with no difference in plasma and myocardial lactate levels among groups. CONCLUSIONS: When used to treat shock in newborn piglets with hypoxia-reoxygenation, milrinone improved cardiac output and carotid flow while maintaining systemic blood pressure. Pulmonary hypertension was not aggravated. Further studies are needed to confirm these findings in asphyxiated neonates.     

飞燕草素葡萄糖苷通过下调微小RNA-106a保护缺氧复氧引起的心肌细胞损伤

目的 探讨飞燕草素葡萄糖苷(DPg)对缺氧复氧(H/R)引起的心肌细胞损伤的影响及其机制.方法 2018年4月至2019年10月,用H9C2细胞建立心肌细胞H/R损伤模型,用常规培养的细胞作为对照组;用终浓度为50μmol/L、100μmol/L、1000μmol/L的DPg培养液处理H9C2细胞24 h,而后进行H/...     

莱菔硫烷对H9C2心肌细胞缺氧/复氧损伤的保护作用

目的:探讨莱菔硫烷(sulforaphane,SFN)对H9C2心肌细胞缺氧/复氧损伤的保护作用。方法:H9C2心肌细胞随机分为3组:正常对照组、缺氧/复氧组(H/R组)和SFN预处理组(10μmol/L SFN预处理细胞12 h后,进行缺氧/复氧处理)。采用倒置显微镜观察各组细胞形态及凋亡程度,CCK8法检测各组H9C2心肌细胞存活率,Western blot法检测血红素氧化酶1(HO-1)、醌氧化还原酶1(NQO1)、Bcl-2和Bax蛋白的表达水平。结果:与正常对照组相比,H/R组和SFN预处理组心肌细胞存活率均降低(P均0.01),HO-1、NQO1、Bcl-2和Bax蛋白表达水平及Bcl-2/Bax均升高(P均0.01)。与H/R组相比,SFN预处理组的细胞生长状态较好,细胞存活率明显提高[(76.00±0.52)%对(55.73±0.43)%,P0.01],HO-1(3.24±0.01对1.86±0.01,P0.01)、NQO1(1.67±0.01对0.95±0.01,P0.01)和Bcl-2(0.70±0.00对0.48±0.01,P0.01)蛋白表达水平及Bcl-2/Bax(1.22±0.01对0.56±0.00,P0.01)均明显升高,Bax蛋白表达水平明显降低(0.57±0.00对0.85±0.01,P0.01)。结论:SFN对H9C2心肌细胞缺氧/复氧损伤有保护作用,可能与促进HO-1、NQO1、Bcl-2蛋白表达,抑制Bax蛋白表达有关。     

Expression of intercellular adhesion molecule-1 (ICAM-1) during hypoxia-reoxygenation by sinusoidal endothelial cells (SECs) in an obstructive jaundice model

Background/Purpose: The aim of the present study was to investigate the expression of intercellular adhesion molecule-1 (ICAM-1) by sinusoidal endothelial cells (SECs) during hypoxia-reoxygenation in an obstructive jaundice model. Methods: Male Wistar rats aged 6 to 8 weeks were assigned to an obstructive jaundice group (group OJ; n= 8) and a sham-operated control group (group S; n= 8). Using flow cytometry, we determined the percentage of ICAM-1-positive cells and the mean fluorescence intensity (MFI) of ICAM-1 per cell in each group during hypoxia-reoxygenation. Results: The percentage of ICAM-1-positive cells in group OJ did not change significantly under successive incubation conditions. However, the MFI in group OJ differed significantly between normoxia and 24-h renormoxia. The present study investigated the function of isolated SECs from rats with obstructive jaundice and hypoxia-reoxygenation. The percentage of ICAM-1-positive cells and the MFI of ICAM-1-positive cells showed quite similar changes caused by hypoxia-reoxygenation stress in group S and Group OJ. The ICAM-1 expression increased with hypoxia-reoxygenation in both normal and OJ SECs, and the amount of ICAM-1, on the whole, increased. Obstructive jaundice increased the percentage of ICAM-1-positive cells from the normal state. Conclusions: The amount of ICAM-1 increased with hypoxia-reoxygenation in obstructive jaundice than normal state. Received: July 13, 2001 / Accepted: November 16, 2001     

缺氧复氧损伤对血管内皮细胞分泌一氧化氮及其合酶活力的影响及辛伐他汀的干预作用

目的:探讨缺氧复氧对血管内皮细胞分泌一氧化氮(NO)及NO合酶(NOS)活力的影响及辛伐他汀的干预作用,并分析其产生作用的可能机制。方法:体外培养人脐静脉内皮细胞株ECV304,使用自制的缺氧小室对ECV304进行缺氧复氧处理。第1实验:仅进行缺氧复氧处理。第2实验:不同浓度的辛伐他汀(0.1、1.0、10.0μmol/L)以及10.0μmol/L辛伐他汀加0.2mmol/L甲羟戊酸预处理ECV30424h后,再行缺氧2h复氧2h处理。硝酸还原酶法及化学比色法分别检测培养液上清中NO的含量及NOS活力。结果:缺氧2h、复氧2、4hECV304分泌NO与对照组相比明显减少,复氧2、4hNOS活力与对照组相比明显下降;经1.0、10.0μmol/L浓度辛伐他汀预处理ECV304后再行缺氧复氧处理则NO含量及NOS活力较对照组明显增加;同时用辛伐他汀和甲羟戊酸预处理后NO含量和NOS较对照组差异无统计学意义。结论:缺氧复氧应激下血管内皮细胞分泌NO功能下降、NOS活力降低;辛伐他汀可以阻止因缺氧复氧刺激导致的NO减少及NOS活力下降,该作用可被甲羟戊酸逆转。