Network pharmacology and subsequent experimental validation reveal the synergistic myocardial protection mechanism of Salvia miltiorrhiza Bge.and Carthamus tinctorius L.

Objective:To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge(S.miltiorrhiza,Dan Shen)and C.tinctorius L.(C.tinctorius,Hong Hua)as an herb pair through network pharmacology and subsequent experimental validation.Methods:Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of s.miltiorrhiza and C.tinctorius as herb pair.Molecular docking was used to verify the binding of the components of these herbs and their potential targets.An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects,which were evaluated using the combination index.Western blot was performed to detect the protein expression of these targets.Results:Network pharmacology analysis revealed 5 pathways and 8 core targets of s.miltiorrhiza and C.tinctorius in myocardial protection.Five of the core targets were enriched in the hypoxia-inducible factor-1(HIF-1)signaling pathway.S.miltiorrhiza-C.tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway.As an upstream gene of the HIF-1 pathway,STAT3 can be activated by the active ingredients cryptotanshinone(Ctan),salvianolic acid B(Sal.B),and myricetin(Myric).Cell experiments revealed that Myric,Sal.B,and Ctan also exhibited synergistic myocardial protective activity.Molecular docking verified the strong binding of Myric,Sal.B,and Ctan to STAT3.Western blot further showed that the active ingredients synergistically upregulated the protein expressionof STAT3.Conclusion:The pharmacodynamic transmission analysis revealed that the active ingredients of S.miltiorrhiza and C.tinctorius can synergistically resist ischemia through various targets and pathways.This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.

Research Progress on Myocardial Protection Strategies

Myocaridial protection aims to salvage myocardium from ischemia and reperfusion injury and to reduce infarct size and its consequences.After more than 30 years of development,the concept of ischemic preconditioning has evolved into"ischemic conditioning",a term that encompasses a number of related endogenous cardioprotective strategies,which can be applied either directly to the heart(ischemic preconditioning or postconditioning)or from afar,for example to a limb(remote ischemic preconditioning,preconditioning,or postconditioning).A variety of cardioprotective therapies have shown promising results in reducing infarct size and improving clinical outcomes in patients with ischemic heart disease.

Sevoflurane Versus Propofol for Myocardial Protection in Patients Undergoing Coronary Artery Bypass Grafting Surgery: a Meta-analysis of Randomized Controlled Trials

Objective To systematically review randomized controlled trials to compare myocardial protection profiles of sevoflurane with propofol in patients undergoing coronary artery bypass grafting （CABG） surgery. Methods Electronic databases were searched to identify all randomized controlled trials comparing sevoflurane with propofol for protecting myocardium in adult patients undergoing CABG surgery. Two authors independently extracted patients＇ perioperative data, including patients＇ baseline characteristics, surgical variables, and outcome data. For continuous variables, treatment effects were calculated as weighted mean difference （WMD） and 95% confidential interval （C/）. For dichotomous data, treatment effects were calculated as odds ratio （OR） and 95% CI. Each outcome was tested for heterogeneity, and randomized-effects or fixed-effects model was used in the presence or absence of significant heterogeneity （Q test P〈0.05）. Sensitivity analyses were done by examining the influence of statistical model on estimated treatment effects. Publication bias was explored through visual inspection of funnel plots of the outcomes. Statistical significance was defined as P〈0.05. Results Our search yielded 13 studies including 696 patients, and 402 patients were allocated into sevoflurane group and 294 into propofol group. There was no significant difference in postoperative mechanical ventilation time, inotropic support, mortality, myocardial infarction, and atrial fibrillation between the two groups （all P〉0.05）. Patients randomized into sevoflurane group had higher post-bypass cardiac index （WMD=0.39, 95% CI： 0.18 to 0.60, P=0.0003）, lower troponin I level （WMD=-0.82, 95% CI：-0.87 to -0.85, P=0.0002）, lower incidence of myocardial ischemia （OR=0.37, 95% CI： 0.16 to 0.83, P=0.02）, shorter ICU and hospital stay length （WMD=-10.99, 95% CI： -12.97 to -9.01, P〈0.00001; WMD=-0.78, 95% CI： -1.00 to -0.56, P〈0.00001, respectively）. Conclusion This meta-analysis has found some evidence showing that sevoflurane has better myocardial protection than propofol in CABG surgery.

Effects of Warm Blood Cardioplegic Solution on Myocardial Protection

To evaluate the effects of warm blood cardioplegic solution on myocardial protection, normothermic induction and terminal perfusion of oxygenated blood cardioplegia in combination with intermittent administration of cold blood cardioplegia during ischemia were studied in an isolated working rat heart model.The experimental protocol consisted of a 120 min cardioplegic arrest followed by 45 min normothermic reperfusion. Myocardial content of adenosine triphosphate (ATP), recovery of the left ventricular function, release of creatine phosphokinase (CPK) and ultrastructure of myocardium were assessed before and after ischemia. The results showed that the hearts preserved with warm blood cardioplegic induction and terminal perfusion had significantly higher levels of ATP,better recovery of cardiac function and lower releases of CPK than those receiving cold blood cardioplegia alone, with myocardial tissue being of generally normal structure. These findings suggest that warm induction and terminal perfusion of blood cardioplegic solution can accelerate myocardial metabolic and functional recovery, preserve high-energy phosphate, reduce myocardial injury and enhance myocardial protection.

Experimental and Clinical Research of Myocardial Protection Effect Using MHBC Perfusion

Objectives To determine themyocardium -protecting effect of medium hypothermalblood cardioplegia (MHBC); further demonstrates thatthe optimal temperature between these hypothermaland normothermic can overcome the disadvantages;and thus discovers a more effective myocardium pro-tecting method. Methods Section 1: 14 mongreldogs (15-20 kg) were randomly divided into twogroups: experimental group and control group; car-dialpulmonary bypass was conventionally instituted,moderate hypothermia blood cardioplegia was used inexperimental group, Blood samples from right atriumtaken for examination of lactate dehydrogenase (LDH)creatine kinase(CK-MB) and Topin Ⅰ(cTn-Ⅰ). speci-mens of left ventricular subendocardial myocardiumwere biopsied to observe changes of ultrastructure.Section 2: 24 patients were randomly divided into twogroups and both groups received two types of treat-ment (same as Section 1) after aorta cross-clamp(ACC). Biochemical index and Clinical observationwere caculated as the indicators. Results In the ex-perimental research, LDH, CK-MB, cTn-Ⅰ were foundincreased afer reperfusion in both groups, but the ex-tent of changes in experimental group is tiny (statisti-cal difference). Compared with control group the effectof MHBC on cardial function is litile; the ultrastruc-ture. of cardiac muscle has no obvious change. In theclinical research, compared the test results of venousblood drawn before CPB, after beating recovery andafter CPB, CBC perfused group (LDH, CK-MB leak-age and cTn-Ⅰ value increase.) compared with MHBCperfused group had no remarkable difference (P>0.05), but at the result of clinical observation: MHBCperfused group had red, soft hearts after cardiac arrestbut CBC perfused group had pale, spasmatic heartsafter cardiac arrest; 0 case in MHBC perfused groupand 3 cases in CBC perfused group had twitch-re-moving beat recovery; 11 cases in MHBC perfusedgroup and 3 cases in CBC perfused group recoveredsinus heart rhythm after surgery; average consumptionof lidocaine was 16.67 (±55.28) mg for MHBC per-fused group and 118.33(±82.65) mg for CBC perfusedgroup (P<0.01) afer surgery; 4 cases in MHBC per-fused group and 11 cases in CBC perfused group suf-fered arrhythmia after surgery. Conclusions As anew myocardium - protecting method, MHBC perfusionin combination with natural body temperature drop ofCPB is worth clinical dissemination and application.

Myocardial Protection during Cardiac Surgery: Warm Blood versus Crystalloid Cardioplegia

Purpose: Prevention of myocardial injury is essential during cardiac surgery. Both crystalloid and blood cardioplegia are popular methods for myocardial protection. Most experimental studies have been in favor of blood cardioplegia. The objective of this study is to determine whether the use of warm blood cardioplegia (BCP) is superior to crystalloid cardioplegia (CCP) by means of myocardial injury markers and clinical outcome parameters. Materials and Methods: In a consecutive series of 293 patients, the first 150 received crystalloid cardioplegia, whereas the next 143 patients received blood cardioplegia. Postoperative myocardial injury was assessed by CTnI and CK-MB. Perioperative morbidity and mortality and clinical outcome parameters (need for inotropic support, ICU and hospital stay) were recorded. An unpaired student t-test was performed to analyse continuous postoperative variables relating to myocardial damage. The presence of possible confounders influencing the CTnI or CK-MB concentrations was tested using a student t-test for continuous variables, for categorical variables ANOVA was used. A final longitudinal model was created for CTnI and CK-MB. CTnI was analyzed by a mixed model with random intercept and slope. For all tests performed, statistical significance was 5%. Results: Both groups were well matched with respect to preoperative variables. No significant difference could be found in maximum postoperative levels of CTnI (8.8 ± 18.4 μg/l in BCP vs 9.6 ± 16.5 μg/l in CCP, p = 0.6455) or CK-MB (19.2 ± 31.0 μg/l in BCP vs 26.4 ± 41.5 μg/l in CCP, p = 0.1209). Nor was there any significant difference in other postoperative variables. Testing treatment effect over time proved only significant influence of the surgical intervention type on CTnI levels in time (p < 0.001). Conclusion: This study could not show significantly higher myocardial injury in the group of patients receiving crystalloid cardioplegia versus warm blood cardioplegia. This suggests that warm blood cardioplegia does not confer superior myocardial protection. Surgical intervention type has an important effect on CTnI concentration in time, while the type of cardioplegia does not.

Efficacy of electroacupuncture on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass:a systematic review and Meta-analysis

OBJECTIVE:To evaluate the efficacy of electroacupuncture(EA)intervention on myocardial protection and postoperative rehabilitation in patients undergoing cardiac surgery with cardiopulmonary bypass(CPB).METHODS:Eight databases,including Pub Med,Embase,the Cochrane Library,Web of Science,Chinese Bio Medical Literature Database,China National Knowledge Infrastructure Database,Wanfang Data,China Science and Technology Journal Database,and two clinical trial registries,were searched.All randomized controlled trials(RCTs)related to EA intervention in cardiac surgery with CPB were collected.Based on the inclusion and exclusion criteria,two researchers independently screened articles and extracted data.After the quality evaluation,RevMan 5.3 software was used for analysis.RESULTS:Fourteen RCTs involving 836 patients were included.Compared with the control treatment,EA significantly increased the incidence of cardiac automatic rebeat after aortic unclamping[relative risk(RR)=1.15,95%confidence interval(CI)(1.01,1.31),P<0.05;moderate].Twenty-four hours after aortic unclamping,EA significantly increased the superoxide dismutase[standardized mean difference(SMD)=0.96,95%CI(0.32,1.61),P<0.05;low],and interleukin(IL)-2[SMD=1.33,95%CI(0.19,2.47),P<0.05;very low]expression levels and decreased the malondialdehyde[SMD=-1.62,95%CI(-2.15,-1.09),P<0.05;moderate],tumour necrosis factor-α[SMD=-1.28,95%CI(-2.37,-0.19),P<0.05;moderate],and cardiac troponin I[SMD=-1.09,95%CI(-1.85,-0.32),P<0.05;low]expression levels as well as the inotrope scores[SMD=-0.77,95%CI(-1.22,-0.31),P<0.05;high].There was no difference in IL-6 and IL-10 expression levels.The amount of intraoperative sedative[SMD=-0.31,95%CI(-0.54,-0.09),P<0.05;moderate]and opioid analgesic[SMD=-0.96,95%CI(-1.53,-0.38),P<0.05;low]medication was significantly lower in the EA group than in the control group.Moreover,the postoperative tracheal intubation time[SMD=-0.92,95%CI(-1.40,-0.45),P<0.05;low]and intensive care unit stay[SMD=-1.71,95%CI(-3.06,-0.36),P<0.05;low]were significantly shorter in the EA group than in the control group.There were no differences in the time to get out of bed for the first time,total days of antibiotic use after surgery,or postoperative hospital stay.No adverse reactions related to EA were reported in any of the included studies.CONCLUSIONS:In cardiac surgery with CPB,EA may be a safe and effective strategy to reduce myocardial ischaemia-reperfusion injury and speed up the recovery of patients after surgery.These findings must be interpreted with caution,as most of the evidence was of low or moderate quality.More RCTs with larger sample sizes and higher quality are needed to provide more convincing evidence.

A method for isolating rat cardiomyocytes by acute enzymatic hydrolysis

Objective: This r esea rch showed a method of isolate rat cardio myocytes though acute enzymatic hydrolysis. Method: Our method was divided into five steps: preparation, heart ext raction, perfusion, mechanical dissociation and pu rifica tion and r ecalcifica ti on. Fir stly, make a preparation, including solutions, equipment and so on. Secondly, after anesthesia satisfactorily, open the chest of rat and take the heart out. Thirdly, transect the aorta and hang the aorta onto the Langendroff perfusion system such that the tip is just distal to the aortic valve. For th, perfuse the hea rt with diges tion solution for 17-20 min afte r Ca2+ free tyrode solution for 5 min. Finally, remove the heart from the system quickly and dissociate mechanically in the KB solution and recalcification after filter. Result: Though the microscope, good-state cardiomyocytes (clear stripes, strong sense of three-dimensional, stick well) can be observed. Conclusion: Those cells can be used to undergo medical electrophysiological experiment, including the study of iron channel.

An injectable alginate/fibrin hydrogel encapsulated with cardiomyocytes and VEGF for myocardial infarction treatment

Myocardial infarction(MI)is one of the typical cardiovascular diseases,which persist as the leading cause of death globally.Due to the poor regenerative capability of endogenous cardiomyocytes(CMs),the transplantation of exogenous CMs becomes a promising option for MI treatment.However,the low retention and survival of transplanted cells still limit the clinical translation of cell therapy.Herein,an alginate/fibrin-based injectable hydrogel was prepared for the delivery of neonatal CMs and an angiogen-esis agent vascular endothelial growth factor(VEGF)locally to the infarcted area of the heart.This hydro-gel combined the specific advantages of alginate and fibrin with proper mechanical properties and cell affinity,showing good biocompatibility to support the retention and integration of the transplanted CMs to the host myocardium.Moreover,the delivered VEGF was favorable for the blood recovery to mitigate the ischemic microenvironment of the infarcted area and thus improved the survival of the transplanted CMs.Intramyocardial injection of this hydrogel to the infarcted area of the heart promoted angiogenesis,inhibited fibrosis,and improved cardiac function,exhibiting great potential for MI treatment.

Maturation of induced pluripotent stem cell-derived cardiomyocytes and its therapeutic effect on myocardial infarction in mouse

Induced pluripotent stem cell-derived cardiomyocytes(iPSC-CMs)have an irreplaceable role in the treatment of myocardial infarction(MI),which can be injected into the transplanted area with new cardiomyocytes(Cardiomyocytes,CMs),and improve myocardial function.However,the immaturity of the structure and function of iPSC-CMs is the main bottleneck at present.Since collagen participates in the formation of extracellular matrix(ECM),we synthesized nano colloidal gelatin(Gel)with collagen as the main component,and confirmed that the biomaterial has good biocompatibility and is suitable for cellular in vitro growth.Subsequently,we combined the PI3K/AKT/mTOR pathway inhibitor BEZ-235 with Gel and found that the two combined increased the sarcomere length and action potential amplitude(APA)of iPSC-CMs,and improved the Ca^(2+)processing ability,the maturation of mitochondrial morphological structure and metabolic function.Not only that,Gel can also prolong the retention rate of iPSC-CMs in the myocardium and increase the expression of Cx43 and angiogenesis in the transplanted area of mature iPSC-CMs,which also provides a reliable basis for the subsequent treatment of mature iPSC-CMs.

Myocardial protection of immature rabbits with an ATP-sensitive K^+ channel opener pinacidil

Objective To investigate the effectiveness of pinacidil,an opener of ATP-sensitive K+ channels,in protecting the myocardium of immature rabbit hearts from ischemic reperfusion injury.Methods Rabbit hearts underwent 30 min of global normothermic ischemia followed by 30 min of reperfusion on the modified Langendorff apparatus.Fifty-two isolated hearts of 3 - 4 week-old immature rabbits were divided into 4 groups randomly.During ischemia,3 different cardioplegic solutions were administered intermittently by infusion every 15 min(20-25 mi each time in all groups).Group 1:control group(n = 13);group 2:Krebs-Henseleit(K-H)solution with potassium(16 mmol/L)(n = 1:3);group 3:K-H solution with potassium(16 mmol/L)and pinacidil(50 μmol/L)(n = 13);group 4:K-H solution with potassium(16 mmol/L),pinacidil(50 μmol/L)and glibenclamide(10 μmol/L)(n = 13).The pre-ischemic and post-ischemic myocardial functions were assessed by the percentage recovery of the left ventricular developed pressure(LVDP);the left ventricular end-diastolic pressure(LVEDP);both the Positive peak and negative peaks of the first derivative of the left ventricular pressures(± dp/dtmax);coronary flow;the level of creatine kinase(CK),lactic dehydrogenase(LDH)and aspartate transcarbamoylase(AST)in coronary sinus venous effluent;and by myocardial ultrastructural changes.Results Before myocardial ischemia,there were no significant differences among the four groups in any of the parameters mentioned above.Post-ischemic recovery of LVDP,LVEDP,± dp/dtmax,coronary flow,the level of CK,LDH and AST,and myocardial ultrastructural changes were better in group 3 than those in the three other groups.Conclusions As a new and effective composition,pinacidil can significantly improve myocardial protection from cardioplegia for immature rabbit hearts.

Myocardial protection during heart surgery in China

Myocardial protection （MP） is the key for cardiopulmonary bypass （CPB） heart surgery. MP during cardiac surgery （CS） aims to preserve myocardial function while providing a bloodless and motionless operating field. Strategies on how to attenuate or prevent post-ischemic myocardial dysfunction that occurs intra-operatively during CS have been discussed for more than half a century. In 1950, Bigelow et al, first reported to decrease myocardial oxygen demand by means of hypothermia. Moreover, Melrose and coworkers2 described the use of electromechanical cardiac arrest induced by potassium infusion, permitting CS to be performed on a non-beating flaccid heart and clear surgical field. The combination of both of these techniques has been the golden standard in MP during surgery until now, allowing surgery with excellent clinical outcome. In 1975, Braimbridge et al introduced a crystalloid solution into clinical practice at St. Thomas Hospital. By the 1980s, blood-based potassium solutions were advocated to further improve MP and to reduce myocardial enzymes release based on the theory that blood would be a superior delivery vehicle for its oxygenating and buffering capacity.Fortunately, the majority of MP strategies now available do allow patients to undergo conventional and complex CS with an operative mortality rate ranging from less than 2% to 4%.

MORPHOMETRIC EVALUATION ON MYOCARDIAL PROTECTION OF COLD CRYSTALLOID VERSUS WARM BLOOD CARDIOPLEGIA

Twenty patients undergoing open-heart valve replacement were divided randomly into two groups in this study; intermittent perfusion of cold crystalloid (St. Thomas Hospital solution) with hypothermic cardiopulmonary bypass (CPB) (hypothermic group) and continuous administration of warm blood cardioplegia with normothermic CPB (normothermic group) respectively. Tissue samples were taken from the right atrium before weaning from CPB and from the right appendage 30 minutes after removal of the cross-clamp. The results of pathological study in these two groups were as follows: the structural alterations were most severe during the ischemic period in the hypothermic group. Damages of the myocardial

Inhibition of TGF-β1 by eNOS gene transfer provides cardiac protection after myocardial infarction

Objective： Endothelial nitric oxide synthase （eNOS） and nitric oxide （NO） have been implicated in protection against myocardial ischemia injury. This study was designed to explore a new method of therapy for myocardial injury by eNOS gene transfection. Methods： A rat model of myocardial infarction （MI） was established by left anterior descending （LAD） coronary artery ligation, eNOS gene in an adenovirus vector was delivered locally into the rat heart and hemodynamic parameters were examined after 3 weeks, Matrix metalloproteinase-2 and 9 （MMP-2, MMP-9） mRNA were measured by reverse transcription polymerase chain reaction （RT-PCR）, and the protein levels of eNOS, caspase-3, and transforming grouth factor 131 （TGF-131） were determined by western blot assay. Results： eNOS gene transfer significantly reduced cardiomyocyte apoptosis and improved cardiac function. In addition, eNOS significantly reduced the mRNA levels of MMP-2 and MMP-9. In the eNOS gene transfected group, the activation of caspase-3 and TGF-β1 were decreased. However, the protection was reversed by administration of the NOS inhibitor, N（o））-nitro-l-arginine methyl ester （L-NAME）. Conclusion： These results demonstrate that the eNOS provides cardiac protection after myocardial infarction injury through inhibition of cardiac apoptosis and collagen deposition, and suppression of TGF-β1.

Effect of a distal protection device on epicardial blood flow and myocardial perfusion in primary percutaneous coronary intervention

Objective: The beneficial effect of percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI) has been well established, but there is the problem of no-reflow phenomenon which is an adverse prognostic factor in primary PCI. In the present study the effect of a distal protection device (PercuSurge GuardWire; GW) on epicardial blood flow and myocardial perfusion was evaluated. Methods and Results: Patients with AMI were randomly divided into 2 groups, the GW and the control groups. The GW group included 52 patients with AMI who underwent primary PCI with GW protection and the control group included 60 patients who underwent primary PCI without GW protection. Epicardial blood flow in the infarct-related artery (IRA) and myocardial perfusion were evaluated according to the thrombolysis in myocardial infarction (TIMI) flow grade and the myocardial blush grade (MBG). We found TIMI score of 3 was obtained significantly more frequently in the GW group (96%) than in the control group (80%). The MBG score of 3 was obtained also significantly greater in the GW group (65%) than in the control group (33%). Conclusion: Primary PCI with GW protection can significantly improve epicardial blood flow and myocardial perfusion.

川芎嗪调控Wnt/β-连环蛋白信号通路对冠心病大鼠心肌细胞凋亡的机制研究

目的:探讨川芎嗪调控Wnt/β-连环蛋白(Wnt/β-catenin)信号通路对冠心病(CHD)大鼠心肌细胞凋亡的影响。方法:选择成年雄性SD大鼠50只,分为对照组、模型组、川芎嗪低、中、高剂量组,各10只。采用高脂饲料喂养加脂肪乳剂灌胃建立CHD大鼠模型,对照组和模型组大鼠尾静脉注射等体积生理盐水,川芎嗪低、中、高剂量腹腔注射川芎嗪,各组均连续治疗3周。干预后检测各组大鼠心功能和血清乳酸脱氢酶(LDH)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)水平,采用HE染色观察心肌组织结构,TUNEL检测心肌细胞凋亡情况,RT-qPCR、Western blot法检测心肌组织Wnt3a、β-cateninmRNA及蛋白表达。结果:模型组大鼠心功能低于对照组和川芎嗪低、中、高剂量组(均P<0.05)。模型组大鼠血清LDH、CK、CK-MB水平高于对照组和川芎嗪低、中、高剂量组(均P<0.05)。对照组心肌细胞大小均等,心肌纤维无肿胀、排列紧密,模型组心肌纤维呈现明显肿胀、断裂,细胞排列无规则,并有大量炎症细胞浸润,川芎嗪低、中、高剂量组心肌损伤逐渐改善,细胞水肿变性减少,仅少量炎性细胞浸润。川芎嗪高剂量组心肌组织中Wnt3a、β-cateninmRNA和蛋白表达低于低、中剂量组(均P<0.05)。结论:川芎嗪能改善大鼠心肌细胞的凋亡,减轻细胞损伤,其机制可能与调节Wnt/β-catenin信号通路相关蛋白表达有关。

过表达BMAL1对脓毒症小鼠心肌损伤的保护作用及机制研究

目的:探究生物钟基因大脑和肌肉芳香烃受体核转运蛋白样蛋白1(BMAL1)过表达对脓毒症小鼠心肌损伤中线粒体自噬途径的影响。方法:将40只小鼠随机分为模型对照组、模型组、阴性对照(NC)组、BMAL1组,每组10只。超声心动图检测小鼠心功能指标左室短轴缩短率(LVFS)、左室射血分数(LVEF)、左室舒张末期内径(LVIDd)、左室收缩末期内径(LVIDs),ELISA检测血清肌酸激酶同工酶(CK-MB)、肌钙蛋白I(cTnI)水平,HE染色观察心肌组织病理变化,TUNEL染色检测心肌细胞凋亡,透射电镜观察心肌线粒体超微结构变化,Western blotting测定心肌组织中微管相关蛋白1轻链3(LC3)、BH3域自噬蛋白(Beclin1)、PTEN诱导激酶(Pink1)、E3泛素-连接酶活性帕金森病蛋白2(Parkin)、电压依赖性阴离子通道蛋白1(VDAC1)表达水平。结果:与模型组和NC组比较,BMAL1组小鼠LVFS、LVEF升高(P<0.05),LVIDd、LVIDs减小(P<0.05),血清中CK-MB、cTnI降低(P<0.05),心肌组织病变得到改善,TUNEL阳性细胞率减少(P<0.05),线粒体损伤减轻,心肌组织中LC3Ⅱ/LC3Ⅰ值升高(P<0.05),Beclin1、Pink1、Parkin、VDAC1蛋白表达上调(P<0.05)。结论:BMAL1过表达能够明显改善脓毒症小鼠心功能,减轻心肌损伤,该作用可能与其促进线粒体自噬有关。

天麻素在心肌缺血再灌注损伤中的作用机制和研究进展

缺血性心血管疾病是一种全球范围内高发的高病死率疾病,目前对于缺血及再灌注对脏器的损伤仍缺乏有效的治疗措施。天麻素作为一种多用于头痛惊厥的重要药物近年来被应用于心肌缺血再灌注损伤(MIRI)的保护治疗。MIRI的保护机制相关研究主要聚焦于细胞死亡、氧化应激、细胞自噬、炎症损伤、钙超载及微循环损伤等。以往临床研究证据表明,天麻素可通过调节细胞自噬作用影响MIRI多种损伤机制,同时,减少梗死面积及修复损伤功能。本文将近年最新有关天麻素和MIRI的文献进行高度凝练,并阐述天麻素在MIRI中发挥作用与调节自噬、恢复细胞稳态强密切相关,以期为临床应用天麻素保护MIRI提供重要的依据和方向。

阿芬太尼预处理通过抑制内质网应激发挥对心肌缺血再灌注损伤的保护作用

目的探究阿芬太尼预处理对心肌缺血再灌注损伤(myocardial Ischemia reperfusion injury,MIRI)大鼠的改善作用及可能存在的分子机制。方法纳入60只雄性SD大鼠,随机将其分为假手术组(Sham组,n=15)、模型组(MIRI组)、阿芬太尼低剂量预处理组(L-alfentanil组,3 mg/kg)及阿芬太尼高剂量预处理组(H-alfentanil组,6 mg/kg),每组15只;除去假手术组,剩余各组大鼠均构建MIRI模型。统计各组大鼠心功能参数[左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)]、血清心肌损伤因子[肌酸激酶同工酶(CK-MB)、肌钙蛋白Ⅰ(cTnI)]水平差异性;双染色法观察大鼠心肌梗死面积;HE染色分析心肌组织病理损伤;TUNEL检测心肌组织凋亡;免疫荧光及Western-blot检测各组大鼠心肌组织内质网应激蛋白(GRP78、CHOP、FAM134B)表达差异性。结果与Sham组相比,MIRI模型建立可以提升LVEDD、LVESD、CK-MB、cTnI水平、心肌梗死面积占比、心肌组织细胞凋亡程度及GRP78、CHOP表达及荧光强度,下调LVEF、LVFS水平及内质网应激蛋白FAM134B表达及荧光强度(P<0.05)。与MIRI组相比,阿芬太尼预处理可以下调LVEDD、LVESD、CK-MB、cTnI水平、心肌梗死面积占比、心肌组织细胞凋亡程度及GRP78、CHOP表达及荧光强度,上调LVEF、LVFS水平及内质网应激蛋白FAM134B表达及荧光强度(P<0.05)。与阿芬太尼低剂量组相比,阿芬太尼高剂量组大鼠LVEDD、LVESD、CK-MB、cTnI水平、心肌梗死面积占比、心肌组织细胞凋亡程度及GRP78、CHOP表达及荧光强度明显降低,LVEF、LVFS水平及内质网应激蛋白FAM134B表达及荧光强度明显提升,体现出剂量依赖性(P<0.05)。结论阿芬太尼预处理具有减轻心肌缺血再灌注大鼠心肌组织病理损伤,减少心肌梗死面积占比及抑制心肌组织细胞凋亡的作用,剂量依赖性明显,其分子机制可能与抑制内质网应激性程度有关,值得临床进一步研究。

Therapeutic and regenerative potential of different sources of mesenchymal stem cells for cardiovascular diseases

Mesenchymalstemcells(MSCs)areidealcandidatesfortreatingmanycardiovasculardiseases.MSCscanmodify the internal cardiac microenvironment to facilitate their immunomodulatory and differentiation abilities,which are essential to restore heart function.MSCs can be easily isolated from different sources,including bone marrow,adipose tissues,umbilical cord,and dental pulp.MSCs from various sources differ in their regenerative and therapeutic abilities for cardiovascular disorders.In this review,we will summarize the therapeutic potential of each MSC source for heart diseases and highlight the possible molecular mechanisms of each source to restore cardiac function.