Formononetin regulates dilated cardiomyopathy-mediated heart failure in rats via HSP90/AKT cardiomyocyte apoptosis and mechanism / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of formononetin (FMN) on cardiomyocyte apoptosis and HSP90/AKT in rats with dilated cardiomyopathy-mediated heart failure. 【Methods】 Echocardiography, ELISA, histological staining, and TUNEL staining were used to observe the protective effect of different doses of FMN on dilated cardiomyopathy-mediated heart failure in rats and the apoptosis of cardiomyocytes. The potential targets of formononetin on dilated cardiomyopathy-mediated heart failure were obtained from TCMSP, DisGeNet, GeneCards, and other databases, the key targets were obtained according to the protein-protein interaction (PPI) network, and the key targets were verified by molecular docking. Western blotting was used to further verify the regulatory role of key targets in the treatment of dilated cardiomyopathy-mediated heart failure with formononetin. 【Results】 Formononetin could reduce the levels of LVIDS, LVIDD, NT-pro BNP, cTn-T, CK, CK-MB, and LDH in rats with dilated cardiomyopathy-mediated heart failure, increase the levels of EF and FS, and reduce the apoptosis of cardiomyocytes. FMN had a strong binding effect on 10 key targets (AKT1, HSP90AA1, CASP3, MAPK1, MMP9, SRC, ALB, HRAS, IGF1, and EGFR) screened by network pharmacology, with HSP90AA1 and AKT1 having the strongest binding effect. Formononetin decreased the expression of HSP90, AKT and downstream CASP3 protein, but increased the expression of p-AKT in myocardial tissue. 【Conclusion】 Formononetin may inhibit the expression of HSP90, promote phosphorylation of AKT to p-AKT, and inhibit the expression of CASP3, thereby reducing the apoptosis of cardiomyocytes and improving myocardial tissue damage, so as to achieve the purpose of treating dilated cardiomyopathy-mediated heart failure.

Levosimendan alleviates coronary microembolization-induced myocardial injury through LOX-1/p38 MAPK pathway / 中华急诊医学杂志

Objective:To study the effect of levosimendan on coronary microembolization (CME)-induced myocardial injury and LOX-1/p38MAPK pathway.Methods:Microspheres were injected into coronary anterior descending branch to construct swine CME model, swine was given levosimendan by continuous intravenous drip for 24 h before modeling, and myocardial-specific overexpression of lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) was achieved through coronary artery injection of adeno-associated virus (AAVs) at 2 weeks before modeling. Then, echocardiography was used to measure cardiac function; HE staining and HBFP staining were used to observe the pathological changes of myocardium and myocardial microinfarction area, respectively; ELISA was used to detect the serum level of cTnI; TUNLE staining was used to detect cardiomyocyte apoptotic index; the LOX-1, Bax, caspase-3 p12, Bcl-2, and p-p38 MAPK protein in myocardial tissue was observed by immunofluorescence method.Results:Compared to the sham group, the LVEF, LVFS, and CO value in the CME group were decreased, while the LVEDd value was increased significantly (all P<0.05); the area of myocardial micro-infarction, serum cTnI level and cardiomyocyte apoptotic rate in the CME group were increased significantly (all P<0.05); the protein levels of Bax, caspase-3 p12, LOX-1, and p-p38 MAPK were increased significantly, while the Bcl-2 level was decreased significantly ( P<0.05). Levosimendan pretreatment significantly improved cardiac dysfunction, reduced the area of myocardial micro-infarction and serum cTnI level, alleviated cardiomyocyte apoptosis, and significantly reduced the LOX-1 and p-p38 MAPK protein expression levels following CME (all P<0.05); while pretreatment with levosimendan and LOX-1 overexpression AAVs simultaneously abolished the effects of pretreatment with levosimendan alone (all P<0.05). Conclusion:Levosimendan alleviates CME-induced myocardial injury through inhibiting cardiomyocyte apoptosis mediated by LOX-1/p38 MAPK signaling pathway.

MiRNA-138 Protecting Cardiomyocyte Apoptosis of Neonatal Rats by Inhibiting JNK/p38 MAPK Pathway / 中山大学学报(医学科学版)

@#【Objective】To explore miRNA-138 protecting cardiomyocyte apoptosis of neonatal rats by inhibiting JNK/ p38 MAPK pathway.【Methods】Thirty-three whole blood samples from patients with acute myocardial ischemia during the period from January 2018 to December 2018 were collected. Thirty- three whole blood samples from healthy people who underwent physical examination in the same period were enrolled as controls. Ischemia/reperfusion（I/R）models of neonatal rats were established. Forty neonatal rats were randomly divided into blank control group（Control），model group（I/R model），negative control group（injected with unordered sequence）and miRNA-138 overexpression group（Ad-miRNA-138），with 10 cases in each group. The expression of miRNA- 138，Bcl2 and Bax mRNA in whole blood and myocardial tissues was detected by qRT-PCR. The levels of rat hemodynamic indexes were detected by electrophysiological recorder. The pathological damages of myocardial tissues were observed by HE staining. The expression of Caspase- 3 in myocardial tissues was detected by immunohistochemistry. The apoptosis of myocardial cells was observed by TUNEL staining. The content of reactive oxygen species （ROS） in myocardial tissues was detected by ELISA. The expression of JNK/p38 MAPK pathway protein was detected by Western blotting. 【Results】 Compared with healthy control，the level of whole blood miRNA- 138 was significantly decreased in patients with acute myocardial ischemia（P<0.05）. Compared with I/R model group，cardiomyocyte gap in Ad-miRNA-138 group was smaller，its arrangement was more uniform，and its structure was more complete. The miRNA- 138 expression，levels of cardiac function indexes such as +dp/dtmax，HR，LVSP and Bcl2/Bax were significantly increased，while number of apoptosis cells，rate of Caspase-3 positive-expression cells， ROS，p-p38/p38，Pc-jun/c-jun and p-JNK1/2/JNK1/2 were significantly down-regulated（P<0.05）.【Conclusion】MiRNA- 138 can inhibit cardiomyocyte apoptosis of rats，and reduce ROS damage by inhibiting JNK/p38 MAPK pathway， thus protecting cardiomyocytes of neonatal rats.

Effect of Baicalin on myocardial cell apoptosis and Akt/AMPK/mTOR signal pathway expression in chronic myocardial failure rats / 医学研究生学报

ObjectiveTo investigate the effect of Baicalin on the apoptosis of cardiomyocytes and the expression of Akt/Amp activated protein kinase (AMPK)/mammalian rapamycin target protein (mTOR) signaling pathway in rats with chronic myocardial failure.MethodsSixty male SD rats of SPF grade were randomly divided into normal group, model group, positive control group and experimental group, with 15 rats in each group. Rats in the positive control group were given 40mg/kg losartan once a day. Rats in the experimental group were given 50mg/kg Baicalin once a day. Rats in the normal group and the model group were given the same dose of saline once a day. Each group was given gavage for 4 weeks. HE staining was used to observe the pathomorphology of cardiomyocytes. The myocardial tissue index (CAI) was detected by in situ end labeling. The expressions of Bax and bcl-2 protein in myocardium were detected by immunohistochemistry. The cardiac function was detected by echocardiography. The expression of Akt, AMPK and mTOR mRNA was detected by RT-PCR.ResultsCompared with the normal group, the apoptosis indexes of the model group, the positive control group and the experimental group were increased (P0.05). Compared with the model group, the expression of Bcl-2 protein was increased and Bax protein was decreased in the positive control group and the experimental group (P0.05). Compared with the normal group, the LVEFs of the model group, the positive control group and the experimental group were decreased, while the LVD and LVS were increased (P0.05).ConclusionBaicalin can alleviate myocardial injury in rats with chronic myocardial failure, and its mechanism may be related to the upregulation of Akt/AMPK/mTOR signaling pathway.

Effect of Baicalin on myocardial cell apoptosis and Akt/AMPK/mTOR signal pathway expression in chronic myocardial failure rats / 医学研究生学报

ObjectiveTo investigate the effect of Baicalin on the apoptosis of cardiomyocytes and the expression of Akt/Amp activated protein kinase (AMPK)/mammalian rapamycin target protein (mTOR) signaling pathway in rats with chronic myocardial failure.MethodsSixty male SD rats of SPF grade were randomly divided into normal group, model group, positive control group and experimental group, with 15 rats in each group. Rats in the positive control group were given 40mg/kg losartan once a day. Rats in the experimental group were given 50mg/kg Baicalin once a day. Rats in the normal group and the model group were given the same dose of saline once a day. Each group was given gavage for 4 weeks. HE staining was used to observe the pathomorphology of cardiomyocytes. The myocardial tissue index (CAI) was detected by in situ end labeling. The expressions of Bax and bcl-2 protein in myocardium were detected by immunohistochemistry. The cardiac function was detected by echocardiography. The expression of Akt, AMPK and mTOR mRNA was detected by RT-PCR.ResultsCompared with the normal group, the apoptosis indexes of the model group, the positive control group and the experimental group were increased (P0.05). Compared with the model group, the expression of Bcl-2 protein was increased and Bax protein was decreased in the positive control group and the experimental group (P0.05). Compared with the normal group, the LVEFs of the model group, the positive control group and the experimental group were decreased, while the LVD and LVS were increased (P0.05).ConclusionBaicalin can alleviate myocardial injury in rats with chronic myocardial failure, and its mechanism may be related to the upregulation of Akt/AMPK/mTOR signaling pathway.

Long non-coding RNA MALAT1 enhances the apoptosis of cardiomyocytes through autophagy inhibition by regulating TSC2-mTOR signaling

BACKGROUND: Our previous study showed that knockdown of long noncoding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) attenuated myocardial apoptosis in mouse acute myocardial infarction (AMI). This study aims to explore whether MALAT1 enhanced cardiomyocyte apoptosis via autophagy regulation and the underlying mechanisms of MALAT1 regulating autophagy. METHODS: Cardiomyocytes were isolated from neonatal mice and then stimulated with hypoxia/reoxygenation (H/R) injury to mimic AMI. The autophagy level was assessed using GFP-LC3 immunofluorescence and western blot analysis of autophagy-related proteins. RNA pull-down and RNA immunoprecipitation (RIP) was performed to analyze the binding of MALAT1 and EZH2. Chromatin immunoprecipitation (ChIP) assay was performed to analyze the binding of TSC2 promoter and EZH2. The cell apoptosis was evaluated using TUNEL staining and western blot analysis of apoptosis-related proteins. RESULTS: H/R injury increased MALAT1 expression in cardiomyocytes. Furthermore, MALAT1 overexpression inhibited, whereas MALAT1 knockdown enhanced the autophagy of cardiomyocytes. Moreover, MALAT1 overexpression recruited EZH2 to TSC2 promoter regions to elevate H3K27me3 and epigenetically inhibited TSC2 transcription. Importantly, TSC2 overexpression suppressed mTOR signaling and then activated the autophagy. Further results showed that MALAT1 inhibited proliferation and enhanced apoptosis of cardiomyocytes through inhibiting TSC2 and autophagy. CONCLUSION: These findings demonstrate that the increased MALAT1 expression induced by H/R injury enhances cardiomyocyte apoptosis through autophagy inhibition by regulating TSC2-mTOR signaling.

Electroacupuncture and moxibustion pretreatments reduce cardiomyocyte apoptosis and autophagy in rats with myocardial ischemia/reperfusion injury / 针刺研究

OBJECTIVE: To observe the effect of "Neiguan" (PC6)-electroacupuncture (EA) or moxibustion (Moxi) pretreatment on myocardial apoptosis and expression of autophagy related proteins light chain (LC) 3-Ⅰ and LC3-Ⅱ in rats with myocardial ischemia/reperfusion injury (MIRI), so as to explore their mechanisms underlying improvement of MIRI. METHODS: Forty SD rats (half male and half female) were randomly divided into sham operation, model, ischemic preconditioning (IP), EA and Moxi groups (n＝8 in each group). EA (10 Hz/50 Hz, 1 mA) or Moxi (ignited moxa stick) was respectively applied to bilateral "Neiguan" (PC6) for 20 min, once daily for 7 days. The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min. The ultrastructural changes and autophagy of myocardial cells were observed by electron microscopy (EM), and the myocardial cellular apoptosis [apoptotic index ＝ (number of apoptotic cells/total number of cardiomyocytes)×100%] was detected by the terminal deoxyribonucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method. The expressions of LC3-Ⅰ and LC3-Ⅱ proteins (markers for autophagy) in myocardial tissue were detected by Western blot. RESULTS: Following MI, EM observation revealed a vague structure of cardiomyocytes and muscular horizontal grain, dissolution of myofibers, mitochondrial swelling, some autophagic vacuoles and autophagic lysosomes at different degrees and surrounded by a double membrane in the model group, these situations were apparently milder in the EA and Moxi groups. The apoptosis index, myocardial LC3-Ⅰ and LC3-Ⅱ protein expression levels, and the ratio of LC3-Ⅱ/Ⅰ were significantly increased in the model group relevant to the sham operation group (P0.05), and no significant differences were observed in the apoptosis index and levels of LC3-Ⅱ and LC3-Ⅱ/Ⅰ between the IP and Moxi groups (P>0.05)．. CONCLUSION: Both EA and moxibustion pretreatments, similar to IP, have a positive role in reducing myocardiocyte apoptosis and regulating autophagy-related protein expression in MIRI rats, which maybe contribute to their protective effects on ischemic myocardium.

Effect of Zhenwu decoction in the treatment of acute heart failure and its influence on cardiac function, cardiomyocyte apoptosis factor and serum level of brain natriuretic peptide / 中国基层医药

Objective@#To explore the clinical effect of Zhenwu decoction in the treatment of acute heart failure(AHF), and its influence on cardiac function, cardiomyocyte apoptosis factor and serum level of brain natriuretic peptide(BNP).@*Methods@#94 patients with AHF were enrolled in the study.According to their treatment time number, 47 cases with odd numbers were selected as control group, who were given west medicine for treatment, while 47 cases with even numbers were selected as observation group, who were given Zhenwu decoction on the basis of control group.The clinical treatment effect of the two groups was evaluated, and the levels of BNP, soluble Fas(sFas), soluble Fas ligand (sFasL), tumor necrosis factor-α(TNF-α), and cardiac function indicators including left ventricular ejection fraction(LVEF), stroke volume(SV), left ventricular end diastole(LVEDd), left ventricular end of systolic diameter(LVEDs) before and after treatment were compared.@*Results@#The total effective rate of the observation group was 91.49%(43/47), which was higher than that of the control group 65.96%(31/47), the difference between the two groups was statistically significant(χ2=8.871, P<0.05). After treatment, the LVEF and SV in the observation group were (52.77±4.41)% and (79.00±6.54)mL, respectively, which were higher than those in the control group[(43.15±3.88)%, (69.22±4.80)mL], while the LVEDd and LVEDs of the observation group were (50.55±3.43)mm, (37.44±2.73)mm, respectively, which were lower than those of the control group[(59.52±4.02)mm, (42.04±3.20)mm], the differences were statistically significant(t=7.454, 7.042, 6.768, 6.890, all P<0.05). After treatment, the levels of sFas, sFasL and TNF-α in the observation group were (2.65±0.30)μg/L, (0.21±0.08)μg/L, (1.20±0.13)ng/L, respectively, which were lower than those of the control group[(3.38±0.37)μg/L, (0.33±0.12)μg/L, (1.48±0.20)ng/L], the differences were statistically significant(t=6.438, 6.703, 6.185, all P<0.05). After treatment, the BNP level in the observation group was (252.77±27.43)ng/L, which was lower than that in the control group[(387.67±41.97)ng/L], the difference was statistically significant(t=6.870, P<0.05).@*Conclusion@#The curative effect of routine treatment of western medicine combined with Zhenwu decoction in the treatment of AHF is exact, which can significantly improve the clinical symptoms and cardiac function, reduce myocardial cell apoptosis and BNP level, and it has high clinical value, which is worthy of popularization and application.

Effect of Astragalus polysaccharide on LPS-induced cardiomyocyte apoptosis by inhibiting NF-κB and JNK signaling pathway / 中国药理学通报

Aim To study the effect of Astragalus polysaccharide (APS) on the apoptosis of cardiomyocytes induced by lipopolysaccharide (LPS) in mice and to explore its mechanism.Methods Kunming mice were treated with APS for 14 days,and intraperitoneal injection of LPS (10 mg · kg-1) was performed to establish cardiomyocyte apoptosis model in vivo,and H9c2 cells were pre-administered with APS.After 30 min,LPS (1 mg · L-1) was incubated for 24 hours to establish a model of cardiomyocyte apoptosis.Cardiac ejection fraction (EF) and left ventricular shortening score (FS) were measured using echocardiography in mice.TUNEL was carried to measure myocardial cell apoptosis.Enzyme-linked immunosorbent assay (ELISA) was employed to measure serum levels of IL-1β,TNF-α.Western blot was used to detect the expression of JNK,NF-κB signaling pathway and Bcl-2 family and caspase-3 protein in vivo and in vitro.Results LPS could significantly inhibit the left ventricular systolic function in mice and promote myocardial cell apoptosis.The levels of IL-1β,TNF-α in serum,JNK,p-JNK,Bax,caspase-3 and the concentration of NF-κB in nucleus were all increased,while the level of Bcl-2 and the concentration of NF-κB,IκB-α in cytoplasm were reduced.APS could significantly inhibit left ventricular systolic weakening in LPS-induced mice and cut down the apoptosis of cardiomyocytes.The secretion of IL-1β,TNF-α decreased to varying degrees.The protein levels of p-JNK,Bax,caspase-3 in myocardium and NF-κB in nucleus were down-regulated,but those of Bcl-2 and NF-κB,IκB-α in cytoplasm were up-regulated,and JNK had no significant change in vivo and in vitro.Conclusion APS ameliorates the LPS-induced apoptosis of myocardial cells by inhibiting NF-κB and JNK signaling pathway in mice.

Efficacy of tirofiban coronary injection combined with atorvastatin in treatment of acute coronary syndrome with arrhythmia / 中国中西医结合急救杂志

Objective To study the efficacy of tirofiban coronary injection combined with atorvastatin in the treatment of patients with acute coronary syndrome (ACS) and arrhythmia. Methods Ninety-two patients with ACS and arrhythmia admitted to Yiwu Central Hospital from January 2017 to May 2018 were enrolled, and they were divided into a control group and a study group by reference to random number table method, with 46 cases in each group. The patients in the control group received routine treatment; the patients in the study group were treated with tirofiban coronary injection combined with atorvastatin on the basis of the treatment in the control group. The changes of myocardial injury markers, blood lipid levels, inflammatory factors, apoptosis indicators and incidence of adverse reactions were observed before and after treatment in the two groups. Results After treatment the levels of serum cardiac troponin T (cTnT), creatine kinase (CK), CK isoenzyme (CK-MB), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), interleukin-6 (IL-6), hypersensitive C-reactive protein (hs-CRP) and C-myc were significantly lower than those before treatment in the two groups, and all the levels in the study group were lower than those in the control group [cTnT (mg/L):1.5±0.5 vs. 3.2±1.0, CK (U/L): 158.6±17.2 vs. 224.1±20.3, CK-MB (U/L): 30.5±11.4 vs. 44.3±9.7, TC (mmol/L):5.0±0.8 vs. 5.6±0.5, LDL-C (mmol/L): 2.0±1.0 vs. 2.7±1.3, IL-6 (mg/L): 86.5±15.2 vs. 131.4±16.3, hs-CRP (mg/L): 4.7±3.3 vs. 7.3±3.6, C-myc: (18.2±8.1)% vs. (23.4±10.3)%], and all the above differences were statistically significant (all P < 0.05). After treatment, the levels of Bcl-2 in the two groups were significantly higher than those before treatment, and the level in the study group was obviously higher than that of the control group [(78.4±12.2)% vs. (69.3±9.7)%, P < 0.05]. The adverse reactions were found in 6 patients in each group, and the incidence of adverse reactions was the same in the two groups [13.0% (6/46) vs. 13.0% (6/46), P > 0.05]. Conclusions Tirofiban coronary injection combined with atorvastatin is markedly effective in the treatment of patients with ACS and arrhythmia, as it can reduce myocardial cell damage, regulate blood lipid levels, inhibit inflammatory response and antagonize cardiomyocyte apoptosis.

Icariside Ⅱ improves left ventricular cardiomyocyte apoptosis in spontaneously hypertensive rats / 中国药理学通报

Aim To observe the effect of icariside Ⅱ(ICS Ⅱ)on cardiomyocyte apoptosis in spontaneously hypertensive rats (SHR)and to explore its possible mechanism. Methods Thirty male 13-week-old SHRs were randomly divided into model group,ICS Ⅱ low, medium,high and positive drug group (n = 6),ho-mologous male Wistar-Kyoto rats as control group (n =6). After a week of adaptive feeding,ICS Ⅱ low,me-dium and high dose groups were given ICS Ⅱ 4,8,16 mg · kg - 1 (ig,qd),and the positive drug group was given losartan 20 mg·kg - 1 . At the same time,the WKY and SHR group were given equal volume double distilled water. After 12 weeks of administration,the blood pressure was measured in rats. Then,the rats were sacrificed and the left ventricles were separated in order to calculate the left ventricular mass index. HE staining was used to observe the pathological changes of the left ventricle,and the apoptosis of the left ventricu-lar myocardium was detected by TUNEL staining. The expressions of Bcl-2 and Bax mRNA in left ventricle were detected by real time RT-PCR,and Bcl-2,Bax and cleaved-caspase-3 protein expressions were detec-ted by Western blot. Results Compared with WKY group,the blood pressure and left ventricular mass in-dex increased in SHR group (P < 0. 05),and the my-ocardial cell arrangement was disordered and the cell hypertrophy and apoptosis were obvious,accompanied by rupture of filament ;the level of Bax mRNA was up-regulated (P < 0. 05),and Bcl-2 mRNA was down-regulated (P < 0. 05 );the expressions of Bax and cleaved-caspase-3 protein were up-regulated (P <0. 05),and the level of Bcl-2 protein was down-regu-lated (P < 0. 05 ),and the ratio of Bax / Bcl-2 in-creased (P < 0. 05). Compared with SHR group,the blood pressure and left ventricular mass index de-creased in ICS Ⅱ middle,high group and the positive drug group (P < 0. 05);moreover,the arrangement of myocardial cells became more orderly,the cell hyper-trophy and the apoptosis of myocardial cells were im-proved;the level of Bax mRNA was down-regulated and Bcl-2 mRNA was up-regulated (P < 0. 05);the expression of Bax and cleaved-caspase3 protein were down-regulated and the level of Bcl-2 protein was up-regulated (P < 0. 05 );the ratio of Bax / Bcl-2 de-creased (P < 0. 05). Conclusions ICS Ⅱ can im-prove the left ventricular cardiomyocytes apoptosis in SHR,and its mechanism is related to the decrease of blood pressure and the inhibition of mitochondrial ap-optosis pathway.

Improvement of Cardiomyocyte Apoptosis Induced by Hydrogen Peroxide via the Intervention of GLP-1 Signal Pathway / 医学研究杂志

Objective To observe the effect of Exendin-4 on the apoptosis of cardiomyocyte induced by H2O2,and approach the relationship between GLP-1 signal pathway and the injury of cardiomyocyte.Methods Cardiomyocytes were isolated and cultured,and were divided into 5 groups.Intercelluar ROS (reactive oxygen species) was measured,and cell apoptosis rate was evaluated by Flow cytometry in different groups.Also expressions of apoptosis-associated proteins (caspase-3) and PI3 K/AKT were evaluated by western blot.Results Compared with H2O2 group,Ex-4 co-incubation decreased the production of intercelluar ROS levels,also improved the cardiomyocyte apoptosis.At the same time,Ex-4 resulted in the alterations in expressions of the caspase-3 and p-AKT/AKT proteins.However,these effects of Exendin-4 were counteracted significantly by the co-incubation of LY294002.Conclusion The interventions of GLP-1 signal pathway can improve cardiomyocyte apoptosis induced by H2O2 incubation,and the mechanisms might partly attribute to the PI3K/AKT system.

Effect of saponins extracted from Panax japonicus on inhibiting cardiomyocyte apoptosis by AMPK/Sirt1/NF-κB signaling pathway in aging rats / 中国中药杂志

To investigate the effects of saponins extracted from Panax japonicus(SPJ) on cardiomyocyte apoptosis in natural aging rats and explore its underlying mechanisms. SD male rats were randomly divided into four groups: young control group, natural aging group, SPJ low dose group and SPJ high dose group, with 10 rats in each group. The rats in natural aging group, SPJ low and high dose groups were respectively treated with normal saline, SPJ 10 and 60 mg•kg-1•d-1 from the beginning of 18 month-old, 6 days per week for 6 months till 24 month-old. Then the animals were sacrificed. Their myocardial morphology changes were observed by using haematoxylin-eoin(HE) staining; cardiomyocyte apoptosis was tested by using Tunel assays; and the protein expression levels of Bcl-2, Bax, IL-1β, TNF-α, AMPK, p-AMPK, Sirt1, and Ac-NF-κB p65 in myocardial tissues of rats were detected by Western blot. The results showed that SPJ could effectively improve the arrangement disorder of myocardial fibers, reduce the infiltration of inflammatory cells and inhibit cardiomyocyte apoptosis in natural aging rats. At the same time, SPJ could significantly inhibit the protein expression of Bax, IL-1β, TNF-α and Ac-NF-κB p65, and increase the expression of Bcl-2, Bcl-2/Bax, p-AMPK/AMPK and Sirt1 in the heart tissues of natural aging rats. SPJ can effectively inhibit cardiomyocyte apoptosis in natural aging rats, and its mechanisms may be related with the regulation of inflammatory reaction by AMPK/Sirt1/NF-κB signaling pathway.

Protective effect of pioglitazone on cardiomyocyte apoptosis in low-dose streptozotocin & high-fat diet-induced type-2 diabetes in rats.

Background & objectives: Cardiomyocyte apoptosis is one of the pathologic phenomena associated with diabetes and related conditions including obesity, insulin resistance and hyperlipidaemia. In the present study, the protective effects of pioglitazone on cardiomyocyte apoptosis was evaluated in experimental diabetes induced by low dose of streptozoticin (STZ) combined with high fat diet (HFD) in rats. Methods: Male Wistar rats (150-200 g) were injected with low-dose STZ (45 mg/kg, i.v., single dose) and orally fed with a HFD (20 g/day/rat) for a period of 28 days and simultaneously treated with pioglitazone (20 mg/kg/p.o.) for a period of 21 days (from 8th day to 28th day). On 29th day blood was collected, serum separated and used for biochemical parameters. Heart tissue was used for cardiomyocyte apoptosis measurement and also for histopathological examination. Results: Pioglitazone treatment resulted in a decrease in cardiomyocyte apoptosis as revealed by a decrease in cardiac caspase-3, lactate dehydrogenase (LDH) levels and DNA fragmentation, and an increase in Na+K+ATPase levels in diabetic rats. Cardiac histology of diabetic control rats showed dense focal fatty infiltration in the myocardial cells whereas normal architecture with regular morphology and well preserved cytoplasm was observed with pioglitazone treatment. Pioglitazone treatment significantly reduced the heart rate, mean arterial blood pressure, body mass index (BMI) and levels of serum glucose, leptin, insulin, HOMA-IR, total cholesterol (TC) and triglycerides (TGs), apoliproprotein-B glycosylated haemoglobin (HbA1c) levels and atherogenic index, and increased the levels of serum high density lipoprotein cholesterol (HDL-C) and cardiac antioxidant enzymes. Interpretation & conclusions: The present study results suggest that pioglitazone possesses cardiac anti-apoptotic potential in diabetic rat model and can be further explored for its use for treatment of diabetic cardiomyopathy.

Expression of Wnt and NCX1 and its correlation with cardiomyocyte apoptosis in mouse with myocardial hypertrophy

OBJECTIVE@#To study the correlation between expression of Wnt and NCX1 and cardiomyocyte apoptosis in mouse with myocardial hypertrophy.@*METHODS@#C57B/16 male mice were given the subcutaneous injection of 1 mg/kg isoprenaline to build the myocardial hypertrophy model. After 14 d of model building, mice were executed by cervical vertebra luxation. The ratio of heart weight/body weight (HW/BW) and heart weight/tibia length (HW/TL) was observed and proved using HE staining that detected the size of cardiomyocytes. 40 male C57B/16 mice were randomly divided into the sham group (normal saline) and model group (isoprenaline), with 20 mice in each group. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was applied to detect the cardiomyocyte apoptosis; while Western blot and immunohistochemistry were employed to detect the expression of Wnt and NCX1. Meanwhile, the correlation between these two proteins and cardiomyocyte apoptosis was explored.@*RESULTS@#Compared with the sham group, the ratio of HW/BW and HW/TL was increased in the model group, as well as the bigger and hypertrophied cardiomyocytes, decreased number and increased apoptosis of cardiomyocytes, and increased positive expression of Wnt3a, Wnt5a and NCX1 in the cardiac muscle tissue. Besides, there was positive correlation between the expression of Wnt and NCX1 and the cardiomyocyte apoptosis.@*CONCLUSION@#The expression of Wnt3a, Wnt5a and NCX1 in mouse with myocardial hypertrophy is increased and positively correlated with the cardiomyocyte apoptosis.

Effects of atorvastatin on release of endothelial microparticles and myo-cardial apoptosis in rats with acute myocardial infarction / 中国病理生理杂志

AIM: To investigate the effect of atorvastatin ( AT) on the release of endothelial microparticles (EMP) and myocardial apoptosis in the rats with myocardial infarction .METHODS: SD male rats (n=24) were ran-domly divided into 3 groups:sham operation ( sham) group , myocardial infarction ( MI) group and MI+AT group.The rat model of acute myocardial infarction was prepared by coronary artery ligation .At 2 h and 24 h after modeling , the pe-ripheral blood was collected to detect creatine kinase-MB (CK-MB) and cardiac troponin T (cTnT).The circulating levels of EMP were measured by flow cytometry .The myocardial apoptosis was detected by terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling (TUNEL) assay.RESULTS: At 2 h after modeling, the level of CK-MB was signifi-cantly increased in MI group compared with sham group , and the level of EMP and the myocardial apoptotic rate were sig-nificantly increased in MI group and MI +AT group compared with sham group .At 24 h after modeling , the level of EMP was significantly increased in MI group compared with sham group .The levels of CK-MB, cTnT, EMP and the myocardial apoptotic rate were significantly decreased in MI +AT group compared with MI group .Moreover , the level of CK-MB in MI group was significantly increased at 24 h compared with that at 2 h after modeling .The levels of CK-MB, cTnT and EMP were significantly decreased in MI +AT group at 24 h compared with those at 2 h after modeling .CONCLUSION: Ator-vastatin may reduce the level of EMP and the myocardial apoptotic rate in the rats with acute myocardial infarction , indica-ting that atorvastatin plays a role in protecting endothelium .

Expression of Wnt and NCX1 and its correlation with cardiomyocyte apoptosis in mouse with myocardial hypertrophy

Objective: To study the correlation between expression of Wnt and NCX1 and cardiomyocyte apoptosis in mouse with myocardial hypertrophy. Methods: C57B/16 male mice were given the subcutaneous injection of 1 mg/kg isoprenaline to build the myocardial hypertrophy model. After 14 d of model building, mice were executed by cervical vertebra luxation. The ratio of heart weight/body weight (HW/BW) and heart weight/tibia length (HW/TL) was observed and proved using HE staining that detected the size of cardiomyocytes. 40 male C57B/16 mice were randomly divided into the sham group (normal saline) and model group (isoprenaline), with 20 mice in each group. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was applied to detect the cardiomyocyte apoptosis; while Western blot and immunohistochemistry were employed to detect the expression of Wnt and NCX1. Meanwhile, the correlation between these two proteins and cardiomyocyte apoptosis was explored. Results: Compared with the sham group, the ratio of HW/BW and HW/TL was increased in the model group, as well as the bigger and hypertrophied cardiomyocytes, decreased number and increased apoptosis of cardiomyocytes, and increased positive expression of Wnt3a, Wnt5a and NCX1 in the cardiac muscle tissue. Besides, there was positive correlation between the expression of Wnt and NCX1 and the cardiomyocyte apoptosis. Conclusion: The expression of Wnt3a, Wnt5a and NCX1 in mouse with myocardial hypertrophy is increased and positively correlated with the cardiomyocyte apoptosis.

The effects of glucocorticoid on myocardial apoptosis in septic rats / 中华急诊医学杂志

Objective To observe the occurrence of myocardial apoptosis and discussing the mechanism of the effects of glucocorticoid on myocardial apoptosis in septic rats in order to provide the rationale for clinical strategy.Methods A total of 60 Wistar rats weighing 230-280 g were randomly (random number) divided into control group and experimental group (n =30 in each group).Cecal ligation and puncture (CLP) was performed in rats to induce sepsis,and Cefoperazone Sodium/Sulbactam Sodium (200 mg/kg) was injected into caudal vein 4 hours after CLP,twice a day.In addition,glucocorticoid was given to rats of experimental group.After rats sacrificed,their left ventricular myocardia were rapidly taken out and myocardial apoptosis rate was measured and the level of Bcl-2 was assayed at 6 h,24 h,and 72 h after CLP.Measured data were analyzed with independent-samples t-test and One-Way ANOVA.Results The rates of myocardial apoptosis in experimental group were obviously lower than those in control groups respectively (F=9.11,t=5.681,P＜0.01) (6ht=11.416,P＜0.01; 24ht=6.217,P＜0.01; 72 h t =3.76,P ＜0.01).The rates of myocardial apoptosis in 24 h in control and 72 h control groups were distinctively higher than those in 6 h control group,respectively (F =13.254,sig =0.000,P ＜0.01 ; sig =0.004,P ＜ 0.01).The rates ofmyocardial apoptosis in group 24 h control were higher than those in 72 h control group (sig =0.039,P ＜ 0.05).The rates of myocardial apoptosis make no difference among experimental group (F =2.488,6/24 h sig =0.132,P ＞ 0.05 ; 24/72 h sig =0.549,P ＞ 0.05 ; 6/72 h sig =0.053,P ＞ 0.05).Conclusions The rate of myocardial apoptosis is peaked at 24 h in sepsis rat,and the rate of myocardial apoptosis can be obviously decreased by administration of glucocorticoid.

Glucose-lowering rate influences cardiomyocyte apoptosis via ERK1/2 pathway / 中华内分泌代谢杂志

Objective To explore the effects of extracellular regulated protein kinase 1/2 (ERK1/2) signal pathway on cardiomyocyte apoptosis and tumor necrosis factor-α (TNF-α) expression at different glucose-lowing rates,and the influence of glucose-lowing rate on cardiomyocyte injury and inflammatory secretion function,as well as its mechanism.Methods Cardiomyocytes of Wistar neonate rat were maintained in medium supplemented with 25 mmol/L glucose for 72 h.Then the medium was changed to different concentrations of glucose and all cells were divided into five groups.Group A was control group whose medium supplemented with 25 mmol/L glucose.Medium of group B,C,D,E was supplemented with 20,15,10,5 mmol/L glucose (glucose-lowing rate was 5,10,15,20 mmol/L) respectively.Survival rate of cardiomyocyte was measured by CCK8 kit.Cardiomyocyte apoptosis was measured by flow cytometry instrument and laser confocal microscope after Annexin V-PI.TNF-α was measured by ELISA.ERK1/2 protein and phosphorylation were measured by Western blot.Cardiomyocyte apoptosis and TNF-α levels were measured again after U0126 was added.Results At the same time point,along with the glucose-lowing rate increased,survival rate of cardiomyocyte in group A was increased and those in group C,D,E were decreased (P＜ 0.05).TNF-α concentration was increased in group B,C,D and decreased in group E.After 24 h,apoptosis rate decreased in group B and increased in group C,D,E (P＜0.05).ERK1/2 phosphorylation level increased in group B,D,and E(P＜0.05).The ERK1/2 phosphorylation level in group B was the lowest.After U0126 was added,survival rates of cardiomyocyte in all groups were increased (P＜0.01) while TNF-α concentrations were decreased (P＜0.05).In every group,survival rate of eardiomyocyte after 48 h was lower than that after 3 h and 24 h,while TNF-α concentration was higher (P＜0.05).Conclusion Influence of glucose-lowering rate for cardiomyocyte apoptosis and TNF-o is caused by ERK1/2 pathway.In the glucose-lowering course,ERK1/2 pathway promotes cardiomyocytes apoptosis and TNF-α secretion is related with not only osmotic pressure,but also ERK1/2 signal pathway activation as well.

PQS attenuates cardiomyocyte apoptosis induced by thapsigargin through inhibiting endoplasmic reticulum stress / 中国病理生理杂志

AIM:To study the effect of Panax quinquefolium saponin (PQS) on cardiomyocyte apoptosis in-duced by thapsigargin ( TG) .METHODS:Primary cultured cardiomyocytes from neonatal SD rats were divided into con-trol group, TG group, PQS (40 mg/L, 80 mg/L and 160 mg/L) +TG group, si-PERK+TG group, and mock+TG group.The cells were treated with 1 μmol/L TG for 24 h to induce apoptosis.The PERK gene in the cardiomyocytes was knocked down by RNAi.The cell viability was detected by CCK-8 assay.Apoptosis was analyzed by flow cytometry.Wes-tern blotting was used to determine the expression of ERS molecules GRP78, CRT, ATF4 and CHOP, anti-apoptosis pro-tein Bcl-2 and pro-apoptosis protein Bax.RESULTS: Compared with control group, TG significantly and the apoptosis, reduced the cell viability (P<0.05), increased the phosphorylation of PERK and eIF2α, increased the expression of GRP78, CRT, ATF4, CHOP and pro-apoptosis protein Bax, and decreased the expression of anti-apoptosis protein Bcl-2 (P<0.05).Compared with TG group, PQS treatment (160 mg/L) significantly reduced the apoptosis and increased the cell viability (P<0.05).All the 3 different concentrations of PQS significantly increased the expression of anti-apoptosis protein Bcl-2 and reduced the expression of pro-apoptosis protein Bax (P<0.05) in a dose-dependent manner.PQS pre-treatment and knockdown of PERK both reduced the protein levels of GRP78, CRT, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP and pro-apoptosis protein Bax, and increased the expression of anti-apoptosis protein Bcl-2 (P<0.05). CONCLUSION:PQS at concentration of 160 mg/L attenuated cardiomyocyte apoptosis induced by TG.PQS had the simi-lar effect as PERK knockdown on cardiomyocyte apoptosis.The mechanism may be associated with inhibiting PERK-eIF2α-ATF4-CHOP pathway of ERS-related apoptosis.