Garlicin Post-conditioning Suppresses Adhesion Molecules in Porcine Model of Myocardial Ischemia-Reperfusion Injury

Objective: To evaluate whether garlicin post-conditioning can attenuate myocardial ischemiareperfusion injury in a catheter-based porcine model of acute myocardial infarction(AMI) by affecting adhesion molecules integrin β1/CD29 and platelet endothelial cell adhesion molecule-1(PECAM-1/CD31). Methods: Twenty-two swine were devided into 3 groups: 6 in a sham-operation group, and 8 each in the model and garlicin groups. AMI porcine model was established in the model and garlicin groups. The distal parts of the left anterior descending coronary artery in the animals of the model and garlicin groups were occluded by dilated balloon for 2 h, followed by reperfusion for 3 h. Garlicin(1.88 mg/kg) was injected over a period of 1 h, beginning just before reperfusion, in the garlicin group. Real-time polymerase chain reaction, immunohistochemistry and Western blot were carried out to detect mRNA and protein expressions of CD29 and CD31 3 h after reperfusion. Results: Hematoxylin-eosin staining showed a better myocardial structure in the garlicin group after reperfusion. Compared to the model group, garlicin inhibited both the mRNA and protein expression of CD29 and CD31 in reperfusion area and no-reflow area(both P0.05). Conclusions: Garlicin post-conditioning induced cardio-protection against myocardial ischemia-reperfusion injury in this catheter-based porcine model of AMI. The cardio-protective effect of garlicin is possibly owing to suppression of production of CD29 and CD31, by inhibition of the m RNA expression of CD29 and CD31.     

Combined use of transmyocardial laser revascularization and endothelial progenitor cells enhances neovascularization and regional contractility in a canine model of ischemic hearts

The purpose of this study was to determine the combined effect of transmyocardial laser re- vascularization （TMLR） and the implantation of endothelial progenitor cells （EPCs） on cardiac function of ischemic hearts in canines. The left anterior descending artery （LAD） was occluded to establish the canine model of acute myocardial infarct （AMI）. Four weeks later, the animals were randomly divided into four groups： TMLR group, in which transmyocardial laser-induced channels were established at the ischemic region; EPCs＋TMLR group, in which EPCs were locally transplanted into laser-induced chan- nels at the ischemic region; EPCs group, in which the EPCs were injected into the ischemic region; con- trol group, in which the AMI animals received neither TMLR nor EPCs. The peripheral blood （50 mL） was sampled in all groups. Mononuclear cells from the peripheral blood were separated and cultured to obtain spindle-shaped attaching （AT） cells in vitro. AT cells were labeled with 1, 1 ＇-dioctadecyl-1 to 3,3, 3＇,3＇-tetramethyl-indocarbocyanine perchlorate （DiI） before injecting into the laser-induced channels or ischemic region. Four weeks after the first operation, TMLR was performed in the TMLR group and EPCs＋TMLR group, and at the same time, the EPCs originating from the AT cells were mixed with cal- cium alginate （CA）. Then the EPCs-CA composites were implanted into myocardial channels induced by laser in the EPCs＋TMLR group, and into the myocardial infarct area in the EPCs group. All dogs underwent echocardiography at second month after LAD occlusion. Finally the samples of myocardium around the LAD were subjected to histochemical and immunohistologic examinations. The results showed there was no significant difference in the diameter of left atrium and ventricle before treatment among all groups （P〉0.05）. Eight weeks after modeling, the regional contractility in the LAD territory in the EPCs＋TMLR group was increased as compared with control group and TMLR group, but there was no significant difference between control group and TMLR group. Neoangiogenesis was observed in the EPCs＋TMLR group, and the fibrosis was seen in the TMLR group. There was no significant dif- ference in neoangiogenesis around the channels induced by laser among EPCs＋TMLR, EPCs and TMLR groups. It was concluded that TMLR combined with EPCs could improve the regional and global cardiac function in AMI, and augment neovascularizaiton in channels of ischemic myocardium induced by laser.     

No-reflow protection and long-term efficacy for acute myocardial infarction with Tongxinluo: a randomized double-blind placebo- controlled multicenter clinical trial (ENLEAT Trial)

Background No-reflow after emergency percutaneous coronary intervention （PCI） for acute ST segment elevation myocardial infarction （STEMI） is related to the severe prognosis. The aim of this study was to evaluate the efficacy of Tongxinluo, a traditional Chinese medicine, on no-reflow and the infarction area after emergency PCI for STEMI.Methods A total of 219 patients （female 31, 14%） undergoing emergency PCI for STEMI from nine clinical centers were consecutively enrolled in this randomized, double-blind, placebo-controlled, multicenter clinical trial from January 2007 to May 2009. All patients were randomly divided into Tongxinluo group （n=108） and control group （n=111）, given Tongxinluo or placebo in loading dose 2.08 g respectively before emergency PCI with asprin 300 mg and clopidogrel 300 mg together, then 1.04 g three times daily for six months after PCI. The ST segment elevation was recorded by electrocardiogram at hospitalization and 1, 2, 6, 12, 24 hours after coronary balloon dilation to evaluate the myocardial no-flow; myocardial perfusion scores of 17 segments were evaluated on day 7 and day 180 after STEMI with static single-photon emission computed tomography （SPECT） to determine the infarct area.Results There was no statistical significance in sex, age, past history, chest pain, onset-to-reperfusion time, Killip classification, TIMI flow grade just before and after PCI, either in the medication treatment during the follow up such as statin, β-blocker, angiotensin converting enzyme inhibitor （ACEI） or angiotensin receptor blocker （ARB） between two groups. There was significant ST segment restoration in Tongxinluo group compared to the control group at 6 hours （（-0.22±0.18） mV vs. （-0.18±0.16） mV, P=0.0394）, 12 hours （（-0.24 ± 0.18） mV vs. （-0.18±0.15） mV, P=0.0158） and 24 hours （（-0.27±0.16） mV vs. （-0.20±0.16） mV, P=0.0021） reperfusion; and the incidence of myocardial no-reflow was also reduced significantly at 24-hour reperfusion （34.3% vs. 54.1%, P=0.0031）. The myocardial perfusion scores of 17 segments evaluated by static SPECT was improved significantly on day 7 and day 180 after STEMI in Tongxinluo group compared to the control group （0.61±0.40 vs. 0.76±0.42, P=0.0109 and 0.51 ±0.42 vs. 0.66±0.43, P=0.0115, respectively）.There was no significant difference in severe adverse events between two groups.Conclusion Tongxinluo as a kind of traditional Chinese medicine could reduce myocardial no-reflow and infarction area significantly after emergency PCl for STEMI with conventional medicine therapy.     

心肌梗死小型猪围术期模型的构建及内源性血红素氧合酶-1的心脏保护作用

Objective To explore and develop a Chinese Miniswine model of myocardial infarction (MI) with balloon occlusion of coronary artery. Methods After anesthesia, 20 Chinese Miniswines were subjected to 20 mm or 25 mm balloon occlusion of the distal part of the second diagnonal branch of left anterior descending (LAD) through femoral artery or carotid artery for the purpose of precondition with 3-4 times of myocardial ischemia. Then, inflated balloon occluded LAD for 120 min. Electrocardiography and blood pressure were monitored. Echocardiography, LDH, CK, CK-MB and coronary angiography were also investigated to confirm acute myocardial infarction (AMI). All pigs were bred for 8 weeks. Results Eighteen out of twenty pigs underwent successful induction of AMI, but one pig died 20 days after operation, two pigs died of ventricular fibrillation during 60-100 rain after operation. AMI was confirmed by dynamic changes of electrocardiography , LDH,CK,CK-MB and further pathology. Regional wall motion abnormalities were found in areas of the superior part of interventricular septum and cardiac apex 1 h after operation by two dimensional echocardiography, repeated echocardiography 3-5 weeks after operation showed wall motion abnormalities in the areas of anterior left ventricular wall, cardiac apex and interventricular septum. Old myocardial infarction (OMI) was confirmed in 17 survival pigs after operation. Conclusions A minimally-invasive swine model of AMI and OMI by using angioplasty catheterization balloon technique is feasible and relatively effective.     

Myocardial autophagy variation during acute myocardial infarction in rats: the effects of carvedilol

Background The loss of cardiac myocytes is one of the mechanisms involved in acute myocardial infarction （AMI）-related heart failure. Autophagy is a common biological process in eukaryote cells. The relationship between cardiac myocyte loss and autophagy after AMI is still unclear. Carvedilol, a non-selective α1-and β-receptor blocker, also suppresses cardiac myocyte necrosis and apoptosis induced by ischemia. However, the association between the therapeutic effects of carvedilol and autophagy is still not well understood. The aim of the present study was to establish a rat model of AMI and observe changes in autophagy in different zones of the myocardium and the effects of carvedilol on autophagy in AMI rats. Methods The animals were randomly assigned to a sham group, an AMI group, a chloroquine intervention group and a carvedilol group. The AMI rat model was established by ligating the left anterior descending coronary artery. The hearts were harvested at 40 minutes, 2 hours, 24 hours and 2 weeks after ligation in the AMI group, at 40 minutes in the chloroquine intervention group and at 2 weeks in other groups. Presence of autophagic vacuoles （AV） in the myocytes was observed by electron microscopy. The expression of autophagy-, anti-apoptotic- and apoptotic-related proteins, MAPLC-3, Beclin-1, Bcl-xl and Bax, were detected by immunohistochemical staining and Western blotting. Results AVs were not observed in necrotic regions of the myocardium 40 minutes after ligation of the coronary artery. A large number of AVs were found in the region bordering the infarction. Compared with the infarction region and the normal region, the formation of AV was significantly increased in the region bordering the infarction （P 〈0.05）. The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the region bordering the infarction. Meanwhile, the expression of apoptotic-related proteins was significantly increased in the infarction region. In the chloroquine intervention group, a large number of initiated AVs （AVis） were found in the necrotic myocardial region. At 2 weeks after AMI, AVs were frequently observed in myocardial cells in the AMI group, the carvedilol group and the sham group, and the number of AVs was significantly increased in the carvedilol group compared with both the AMI group and the sham group （P 〈0.05）. The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the carvedilol group compared with that in the AMI group, and the positive expression located in the infarction region and the region bordering the infarction. Conclusions AMI induces the formation of AV in the myocardium. The expression of anti-apoptosis-related proteins increases in response to upregulation of autophagy. Carvedilol increases the formation of AVs and upregulates autophagy and anti-apoptosis of the cardiac myocytes after AMI.     

电针对局灶性脑缺血再灌注大鼠脑细胞凋亡及神经功能恢复的影响

Objective To investigate the influence of acpuncture on free calcium in rat brain cells after focal cerebral ischemia reperfusion.Methods 145 adult male SD rats were randomly divided into control group,simple ischemia reperfusion group and acupuncture with ischemia reperfusion group.The middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established by the modified Longa occlusion method. ①The part of free calcium in rat brain cells,focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 15 min.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 3h, 6h and 12h, the rat was killed and its brain cells were made into single cell suspension,marked by Fluo-3/AM.The fluorescence optical density was recorded by laser confocal microscopy.②The part of nerve functional reconstruction, focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 12 hours.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 7 d, 14 d,30 d,60 d and 90 d, the rat was forced to detect it's strength of the dog.Results ①Free calcium in rats of acupuncture therapy group(6h:10.96±1.18;2h:20.9±4.37) was significantly less than that in control group in 6 h and 12 h after reperfusion (6 h: 16.87 ± 3.56,12 h: 34.10 ±1.06)(P<0.05).②The dog in rats of acupuncture therapy group was significantly more than that in control group in 7 d, 14 d after reperfusion (P＜ 0.05 ).No difference of the dog was detected in 30 d ,60 d and 90 d after reperfusion between the two groups.Conclusion Acupunture could decreases the concentration of free calcium and the expression of Caspase-3 mRNA in rat brain cells after focal cerebral ischemia reperfusion, and it can facilitate the recovery of nerve function.     

Effects of Ethyl Pyruvate on Myocardial Apoptosis and Expression of Bcl-2 and Bax Proteins after Ischemia-reperfusion in Rats

In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion （I/R） in vitro and the expression of Bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups （n=8 in each group）： control group was perfused for 120 min. In the I/R group, after 30 min stabilization the injury was induced by 30 min global ischemia followed by 60 min reperfusion. Ethyl pyruvate （EP） group was set up with the same protocol as I/R group except that it was supplied with 2 mmol/L EP 15 rain before ischemia and throughout reperfusion. Myocardial malonaldehyde （MDA） content was measured. Myocardial apoptotic index （AI） was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling （TUNEL） method. The expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of Bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of Bcl-2 protein was up-regulated in EP group （P〈0.05）. These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating Bcl-2 and down-regulating Bax proteins.     

Tacolimus postconditioning alleviates apoptotic cell death in rats after spinal cord ischemia-reperfusion injury via up-regulating protein-serine-threonine kinases phosphorylation

The effects of tacrolimus postconditioning on protein-serine-threonine kinases （Akt） phos- phorylation and apoptotic cell death in rats after spinal cord ischemia-reperfusion injury were investi- gated. Ninety male SD rats were randomly divided into sham operation group, ischemia-reperfusion group and tacrolimus postconditioning group. The model of spinal cord ischemia was established by means of catheterization through femoral artery and balloon dilatation. The spinal cord was reperfused 20 min after ischemia via removing saline out of balloon. The corresponding spinal cord segments were excised and determined for Akt activity in spinal cord tissue by using Western blotting at 5, 15, and 60 min after reperfusion respectively. Spinal cord tissue sections were stained immunohistochemically for detection of the phosphorylated Akt expression at 15 min after reperfusion. Flow cytometry was applied to assess apoptosis of neural cells, and dry-wet weights method was employed to measure water content in spinal cord tissue at 24 h after reperfusion. The results showed that the activities of Akt in tarcolimus postconditioning group were significantly higher than those in ischemia-reperfusion group at 5, 15, and 60 min after reperfusion （P〈0.05, P〈0.01）. The Akt activities reached the peak at 15 min after reperfu- sion in ischemia-reperfusion group and tacrolimus postconditioning group. The percentage of apoptotic cells and water content in spinal cord tissue were significantly reduced （P〈0.01） in tacrolimus postcon- ditioning group as compared with those in ischemia-reperfusion group at 24 h after reperfusion. It is concluded that tacrolimus postconditioning can increase Akt activity in spinal cord tissue of rats, inhibit apoptosis of neural cells as well as tissue edema, and thereby alleviate spinal cord ischemia-reperfusion injury.     

电针对局灶性脑缺血再灌注大鼠脑细胞凋亡及神经功能恢复的影响

Objective To investigate the influence of acpuncture on free calcium in rat brain cells after focal cerebral ischemia reperfusion.Methods 145 adult male SD rats were randomly divided into control group,simple ischemia reperfusion group and acupuncture with ischemia reperfusion group.The middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established by the modified Longa occlusion method. ①The part of free calcium in rat brain cells,focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 15 min.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 3h, 6h and 12h, the rat was killed and its brain cells were made into single cell suspension,marked by Fluo-3/AM.The fluorescence optical density was recorded by laser confocal microscopy.②The part of nerve functional reconstruction, focal cevebral ischemia model of rats were made by thread locking up the blood vessel for 12 hours.30 min later after reperfusion, the Baihui and Shuigou Point in Du meridian were acupunctured electrically 30 min.After 7 d, 14 d,30 d,60 d and 90 d, the rat was forced to detect it's strength of the dog.Results ①Free calcium in rats of acupuncture therapy group(6h:10.96±1.18;2h:20.9±4.37) was significantly less than that in control group in 6 h and 12 h after reperfusion (6 h: 16.87 ± 3.56,12 h: 34.10 ±1.06)(P<0.05).②The dog in rats of acupuncture therapy group was significantly more than that in control group in 7 d, 14 d after reperfusion (P＜ 0.05 ).No difference of the dog was detected in 30 d ,60 d and 90 d after reperfusion between the two groups.Conclusion Acupunture could decreases the concentration of free calcium and the expression of Caspase-3 mRNA in rat brain cells after focal cerebral ischemia reperfusion, and it can facilitate the recovery of nerve function.     

IMPLANTATION OF AUTOLOGOUS BONE MARROW MONONUCLEAR CELLS INTO ISCHEMIC MYOCARDIUM ENHANCES CORONARY CAPILLARIES AND SYSTOLIC FUNCTION IN MINISWINE

Objective To investigate the therapeutic effectiveness of intracoronary implantation of autologous bone marrow mononuclear cells （BM-MNC） in miniswine model of reperfused myocardial infarction.
Methods Sixteen miniswine myocardial ischemic reperfusion injury models made by ligation of the distal one third segment of left anterior descending artery for 90 minutes were randomized into 2 groups. In BM-MNC group （n = 9）, （3.54 ± 0.90）× 10^8 BM-MNC were intracoronary injected, and in the control group （n = 7）, phosphate buffered saline was injected by the same way. Echocardiographic and hemodynamic results, vessel density, and myocardial infarction size were evaluated and compared before and 4 weeks after cell transplantation.
Results In BM-MNC group, there were no differences between before and 4 weeks after transplantation in aspects of left ventricular ejection fraction （LVEF）, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, or ＋dp/dtmax. In control group, LVEF, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, and ＋dp/dtmax decreased significantly 4 weeks after transplantation （P 〈 0.05）. Left ventricular end-diastolic pressure and -dp/dtmax did not change significantly before and after cell transplantation in both groups. Capillary density in BM-MNC group was greater than that in control group [（13.39 ± 6.96）/high power field vs. （3.50 ± 1.90）/high power field, P 〈 0.05]. Infarction area assessed by tetrazolium red staining and the infarction percentage decreased in BM-MNC group compared with those in control group （P 〈 0.05）.
Conclusions Transplantation of BM-MNC into myocardium with ischemic reperfusion injury increases capillary density and decreases infarction area. It has significantly beneficial effect on cardiac systolic function rather than on diastolic function.     

抗血小板药物对猪急性心肌梗死再灌注后无再流的影响

目的评价抗血小板药物防治猪急性心肌梗死(AMI)再灌注后无再流的作用。方法中华小型猪32只随机分成(1)对照组,(2)氯吡格雷与阿司匹林合用组(氯吡格雷第一天300mg/d,以后3mg·kg-1·d-1,阿司匹林10mg·kg-1·d-1,每天喂药一次,共3天),(3)替罗非斑治疗组(以15μg/kg静脉推注后继以0.5μg·kg-1·min-1的剂量持续静点直至实验结束)和(4)假手术组,每组8只。对照组和两治疗组行冠状动脉结扎3h,松解1h制备AMI再灌注模型。AMI前、后和再灌注后均行血流动力学测定和心肌声学造影(MCE)检查,最终行病理学分析。结果替罗非斑和氯吡格雷与阿司匹林合用组均使血小板最大聚集率显著降低(给药前分别为46.8%和45.7%,给药后分别为12.9%和14.3%)。与对照组相比,替罗非斑可显著改善心功能(P<0.05～0.01),冠脉血流量增加(对照组45.8%;替罗非斑组73.2%,P<0.01),减少无再流结扎区心肌范围(LA)(MCE对照组LA78.5%;病理染色测定LA为82.3%;替罗非斑组:MCE染色测定LA22.8%病理染色测定LA为23.2%(P<0.01),减少无再流结扎区心肌坏死范围(NA)(对照组:98.5%;替罗非斑组:89.2%LA,P<0.05)。然而,氯吡格雷与阿司匹林合用组血流动力学指标、冠脉血流量、无再流范围及心肌坏死范围与对照组相比差异无统计学意义(P均>0.05)。结论替罗非斑能有效地防治心肌梗死再灌注后无再流,而氯吡格雷与阿司匹林合用则完全无效。     

A Clinical study of garlicin in treating acute cerebral infarction

METHoDSPatientsAmongtheonehundredandfifty-foura-cutecerebralinfarctionpatients(nomorethan3daysaftertheonsetofthedisease)diagnosedaccordingtothediagnosticstandardformu1atedintheSecondSessionofNationalConferenceonCerebrovascularDiseases,1986(l),8Oweremaleand74female,aged6O.86111.23years.Thirty-sixpersons,22malesand14fe-males,aged57.47I6.6Oyears,confirmedtobehealthybyX-raychestfluoroscopy,electro-cardiogram,examinationofliverfunctionandrenalfunction,androutineexaminationofblood,urineandstoo…     

血府逐瘀胶囊对急性心肌梗死再灌注后心肌及内皮素-1、一氧化氮／一氧化氮合酶体系影响的实验研究

目的：观察血府逐瘀胶囊对急性心肌梗死（acute myocardial infarction．AMI）再灌注后心肌组织及一氧化氮（nitric oxide,NO）／一氧化氮合酶（nitric oxide synthase．NOS）体系和内皮素-1（endothelin-1，ET-1）的影响，探讨其改善血管内皮功能的机制。 方法：Yorkshire猪45只，随机分成假手术组、模型对照组和血府逐瘀胶囊治疗组。开胸结扎左冠状动脉前降支90min后，松解2h制备AMI再灌注模型。测定造模前后和再灌注后血清ET-1和NO的含量；冠状动脉造影观察心肌血流恢复情况；Western blot分析心肌组织内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）和ET—1蛋白表达；病理学分析心肌组织形态变化。 结果：血府逐瘀胶囊组造模后和再灌注后血ET-1水平低于模型组（P〈0．01），而NO表达水平明显高于模型组（P〈0．01）。冠状动脉造影提示治疗组冠状动脉血流通畅情况优于对照组，但结果没有统计学意义（P=0．253）。Western blot显示血府逐瘀胶囊组eNOS蛋白表达较模型组有所提高。ET-1蛋白表达则有所下降（P〈0．01）。HE染色及微血管密度分析显示与模型对照组相比，血府逐瘀胶囊组毛细血管扩张程度明显减轻。心肌组织损伤明显改善，微血管密度明显增加。 结论：内皮细胞受损可能是无再流现象发生的重要机制之一．血府逐瘀胶囊可能通过调控NO和ET-1基因表达来达到对AMI的治疗作用。     

地尔硫卓对猪急性心肌梗死再灌注后无再流的影响

目的评价地尔硫卓防治猪急性心肌梗死(AMI)再灌注后无再流的作用。方法实验动物随机分成对照组、地尔硫卓组(2mg/min冠脉内)和假手术组。前两组行冠状动脉结扎3h,松解1h制备AMI再灌注模型。AMI前、后和再灌注后均行血流动力学测定和心肌声学造影(MCE)检查,最终行病理学分析。结果与AMI前相比,对照组AMI后3h、左室收缩压(LVSP)、心排量(CO)和左心室内压最大收缩和舒张变化速率(±dp/dtmax)均显著下降(均P<0.05,P<0.01),左室舒张末压(LVEDP)显著升高(P<0.01);再灌注后1h仅LVSP显著恢复(P<0.05),然而±dp/dtmax继续显著下降(P<0.05);而地尔硫卓组AMI后3h各项指标变化与对照组相同;但再灌注后1h LVEDP、±dp/dtmax和CO均显著恢复(均P<0.05),且比对照组更显著(均P<0.05)。对照组MCE和病理染色所测的冠脉结扎区心肌范围(LA)高度一致(P>0.05),再灌注后无再流范围(ANR)分别为78.5%和82.3%,心肌坏死范围(NA)占LA的98.5%;而地尔硫卓组两方法所测LA均与对照组相当(均P>0.05),但ANR仅分别为20.6%和19.9%,NA又与对照组差异无统计学意义(P>0.05)。对照组再灌注即刻和再灌注后1h冠脉血流量仅占AMI前的45.8%和50.6%(均P<0.01),而地尔硫卓组冠脉血流量分别提高到80.4%和79.3%,比对照组增加均有统计学意义(均P<0.01)。结论     

Synergistic Protection of Danhong Injection (丹红注射液) and Ischemic Postconditioning on Myocardial Reperfusion Injury in Minipigs

Objective: To explore the synergistic protection of Danhong Injection (丹红注射液,DHI) and ischemic postconditioning on myocardial reperfusion injury in minipigs.Methods: Acute myocardial infarction model was made by balloon occlusion in left anterior descending coronary artery (LAD) of minipigs,and then postconditioning was simulated through inflation/deflation of the angioplasty balloon.Minipigs were divided into four groups: the sham operation group (SH group),the ischemia/reperfusion group (I/R group),the ischemic postconditioning group (POC group) and DHI combined with ischemic postconditioning group (PAD group,DHI 20 mL through ear vein),six in each group.After 24-h continuous observation,myocardial infarction size was assessed by triphenyltetrazolium staining (TTC).Morphological changes of ischemic myocardium were observed by light microscopy,and cardiomyocyte ultrastructure was studied with electron microscopy.The superoxide dismutase (SOD) and malondialdehyde (MDA) activity in heart homogenates were measured by a biochemical method.Results: The myocardial infarction size was smaller in the POC group than in the I/R group (0.26±0.02 vs.0.37±0.09,P0.05),and the PAD group (0.14±0.08) displayed a significantly reduced infarction size relative to the I/R group (P0.01) and POC group (P0.05).The damage of myocardial tissue was severe in the I/R group shown by light and electron microscopy: myocardial fibers disorder,sarcoplasmic dissolution,myofilament fracture,mitochondria swelling and even vacuolization formation and a large number of inflammatory cell infiltrations.Compared with the I/R group,reduction of reperfusion injury in the PAD group included more orderly arranged myocardial fibers,less infiltration of inflammatory cells and maintenance of mitochondrial integrity.Compared with the I/R group,the damage of myocardial tissue in the POC group was improved,but not as significant as that in the PAD group.SOD levels in the POC group and the PAD group were significantly higher than those in the I/R group (96.96±13.43,112.25±22.75 vs.76.32±10.63,P0.05),and MDA was significantly lower in the POC group and the PAD group compared to the I/R group (1.27±0.19,1.09±0.21 vs.1.47±0.16,P0.05).Conclusion: DHI and ischemic postconditioning show a synergistic cardioprotection on myocardial reperfusion injury in minipigs.     

中药通心络对猪急性心肌梗死再灌注后无再流的影响

目的评价通心络防治猪急性心肌梗死(AMI)再灌注后无再流的作用。方法中华小型猪40只随机分成5组,每组8只:(1)AMI模型对照组(对照组),(2)通心络小剂量治疗组(0·05g·kg-1·d-1),(3)通心络中剂量治疗组(0·2g·kg-1·d-1),(4)通心络大剂量治疗组(0·5g·kg-1·d-1),(5)假手术组。通心络各组预给药3d后行冠状动脉结扎180min,松解60min制备AMI再灌注模型。AMI前、后和再灌注后均行血流动力学测定,心肌声学造影(MCE)检查和病理学分析。结果(1)与AMI前相比,对照组AMI后180min左室收缩压(LVSP)、心排量和左心室内压最大收缩和舒张变化速率(±dp/dtmax)均显著下降(P<0·05或P<0·01),左室舒张末压(LVEDP)显著升高(P<0·01);再灌注后60min仅LVSP显著恢复(P<0·05),然而±dp/dtmax继续显著下降(P<0·05)。小、中和大剂量通心络组AMI后180min各项指标变化与对照组相同;再灌注后60min仅大剂量通心络组LVEDP、±dp/dtmax和心排量均显著恢复(均P<0·05),且显著好于对照组(均P<0·05)。(2)对照组MCE和病理染色所测的冠脉结扎区心肌范围(LA)高度一致(P>0·05),再灌注后无再流区范围分别为78·5%和82·3%,心肌坏死面积占结扎区心肌面积的98·5%。3个通心络组LA与对照组相当(均P>0·05),但MCE和病理染色所测无再流范围在中及大剂量通心络组分别为41·1%、42·4%和24·1%、25·0%,坏死心肌范围分别为90·2%及81·2%,均显著小于对照组和小剂量组(P<0·05或P<0·01),大剂量组也显著小于中剂量组(P<0·05或P<0·01)。(3)对照组再灌注后即刻和再灌注后60min冠脉血流量仅占AMI前的45·8%和50·6%(均P<0·01),而大剂量通心络组冠脉血流量分别占AMI前的76·0%和73·5%,均显著高于对照组(均P<0·01)。结论通心络能有效地防治心肌梗死再灌注后无再流,缩小梗死面积;中剂量有效,大剂量更好。     

Effects of Qarlicin on unstable angina pectoris and its relationship with blood lipid and granule membrane protein-140

Objective: To observe the clinical effects of Garlicin on unstable angina pectoris (UAP) and explore how the Garlicin’s effects vary among syndromes as defined by traditional Chinese medicine (TCM).Methods: Fifty-five patients with UAP were randomly divided into the Garlicin group (34 patients) and the control group (21 patients). Each patient was classified according to TCM Syndrome Differentiation as having Cold Syndrome type, Heat Syndrome type, severe blood stasis (SBS) type, and mild blood stasis (MBS) type of UAP. Garlicin 60 mg or nitroglycerin 5 mg was given to the two groups respectively by intravenous drip for 10 days as one therapeutic course. The curative effect was evaluated by symptomatic changes and electrocardiogram. The effective rates as well as indexes such as blood lipid, lipoprotein, apolipoprotein, and granule membrane protein-140 (GMP-140) were compared between groups and types.Results: Garlicin and nitroglycerin group did not differ significantly in effective rate, while that of Garlicin group was higher for the Cold Syndrome type than that of Heat Syndrome type (P < 0. 01). The high density lipoprotein/low density lipoprotein ratio and apolipoprotein A-I level rose markedly in the former type (P < 0.05), while an opposite trend was seen in the Heat Syndrome type. Garlicin was more effective in the SBS type than that in the MBS type, and it markedly decreased GMP-140 in the MBS type.Conclusions: Garlicin is effective in UAP, especially the Cold Syndrome and SBS types. Its mechanism may involve improving blood lipid levels and inhibiting platelet activation.     

Allicin Improves Cardiac Function by Protecting against Apoptosis in Rat Model of Myocardial Infarction

Objective:To study the effects of allicin on cardiac function and underlying mechanism in rat model of myocardial infarction(MI).Methods:Ninety-four Wistar rats were randomly assigned to 6 groups(n=14–16 per group):sham control group[underwent thoracotomy without left anterior descending(LAD)occlusion and only received an injection of the same amount of citrate buffer],MI control group(subjected to LAD occlusion and only received an injection of same amount of citrate buffer),positive control group(subjected to LAD occlusion and received an injection of diltiazem hydrochloride at the dose of 1.5 mg/kg),and MI+allicin groups(subjected to LAD occlusion and received an injection of allicin at the doses of 1.2,1.8,and 3.6 mg/kg).All of the drugs were administered intraperitoneally daily for 21 days.The infarct area was measured by myocardial staining.Hematoxylin-eosin staining was used to observe the pathological changes.Cardiac function parameters were assessed by echocardiography.The myocardial apoptotic index was estimated by terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling staining.The expression of Bax and Bcl-2 were detected by quantificational real-time polymerase chain reaction and Western blot.Results:Treatment with allicin could attenuate the myocardial infarct area(P0.05)and relieve the changes of the myocardium.The left ventricular anterior wall diastolic and systolic thicknesses were increased in the allicin-treated groups(P0.05),while there was no significant difference in the left ventricular posterior wall diastolic and systolic thickness(P0.05).The left ventricular internal diameter in systole,ejection fraction,fractional shortening,and stroke volume were dramatically elevated in allicin-treated rats(P0.05).Allicin dose-dependently reduced creatine kinase and lactate dehydrogenase levels(P0.05).The myocardial apoptotic index was also markedly lowered,and Bax expression was significantly decreased,whereas Bcl-2 expression exhibited an opposite trend in allicin-treated rats(P0.05).Conclusion:Allicin appears to exert a cardioprotective effect that may be linked to blocking Bcl-2/Bax signaling pathway-denpendent apoptosis,further improving cardiac function.     

Huoxue Anxin Recipe(活血安心方)Promotes Myocardium Angiogenesis of Acute Myocardial Infarction Rats by Up-Regulating miR-210 and Vascular Endothelial Growth Factor

Objective: To investigate the microR NAs(miR NAs) expression profile of acute myocardial infarction(AMI) rats and the regulating effects of Huoxue Anxin Recipe(活血安心方, HAR) on angiogenesis-related miR NAs and genes. Methods: Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table: sham, AMI, and AMI+HAR groups(15 in each group). AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI+HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miR NAs expression profile was detected by quantitative realtime polymerase chain reaction(qR T-PCR). The mR NA and protein expressions of vascular endothelial growth factor(VEGF), and a target gene of miR-210 was further detected by qR T-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining. Results: Compared with the sham group, ejection fraction(EF) and fractional shortening(FS) values were decreased significantly in the AMI group(P0.01), while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI+HAR group, EF and FS values were increased significantly(P0.05 vs. AMI group), and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miR NAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miR NAs in the AMI+HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI+HAR group. The relative mR NA and protein expressions of VEGF were decreased significantly in the AMI group(P0.05 vs. sham group), and increased significantly in the AMI+HAR group(P0.01 vs. AMI group). CD31 expression area and optical intensity were decreased significantly in the AMI group(P0.05 vs. sham group), and increased significantly in the AMI+HAR group(P0.01 vs. AMI group). Conclusions: HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.