Emerging protective roles of shengmai injection in septic cardiomyopathy in mice by inducing myocardial mitochondrial autophagy via caspase-3/Beclin-1 axis

Mammalian Meckel´s cartilage is a temporary structure associated with mandible development. Notably, its elimination is not executed by apoptosis, and autophagy was suggested as the major mechanism. Simultaneous reports point to pro-apoptotic caspases as novel participants in autophagic pathways in general. The aim of this research was to find out whether activation of pro-apoptotic caspases (-2, -3, -6, -7, -8 and -9) was associated with autophagy of the Meckel´s cartilage chondrocytes. Active caspases were examined in serial histological sections of mouse mandible using immunodetection and were correlated with incidence of autophagy based on Beclin-1 expression. Caspase-2 and caspase-8 were found in Beclin-1 positive regions, whereas caspase-3, -6, -7 and -9 were not present. Caspase-8 was further correlated with Fas/FasL and HIF-1alpha, potential triggers for its activation. Some Fas and FasL positivity was observed in the chondrocytes but caspase-8 activation was found also in FasL deficient cartilage. HIF-1alpha was abundantly present in the hypertrophic chondrocytes. Taken together, caspase-8 activation in the Meckel´s cartilage was demonstrated for the first time. Caspase-8 and caspase-2 were the only pro-apoptotic caspases detected in the Beclin-1 positive segment of the cartilage. Activation of caspase-8 appears FasL/Fas independent but may be switched on by HIF-1alpha.

Download Full-text

Induction of apoptosis and autophagy in T-lymphocytes of patients with Systemic Lupus Erythematosus

Kazan medical journal ◽

10.17816/kmj2020-347 ◽

2020 ◽

Vol 101 (3) ◽

pp. 347-355

Author(s):

Y V Skibo ◽

A R Fathullina ◽

B R Ibragimov ◽

S N Abramov ◽

R R Ismagilova ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

T Lymphocytes ◽

Lupus Erythematosus ◽

Peripheral Blood ◽

Caspase 3 ◽

Immunomagnetic Separation ◽

Beclin 1 ◽

Systemic Lupus ◽

Software Environment ◽

Average Value

Aim. To analyze the expression of key apoptosis (Bcl-2, caspase-3) and autophagy (Beclin 1, Vps34, p62 and LC3) proteins regulators in peripheral blood T-lymphocytes of patients with systemic lupus erythematosus. Methods. The object of the study was peripheral blood T-lymphocytes of healthy donors and patients with systemic lupus erythematosus. To obtain T cells, we used the immunomagnetic separation method. Protein expression was analyzed using the Western blot method. Statistically analyzing the results was performed using the R software environment. The data was represented using boxplots. Groups were compared using the MannWhitney test. Results. According to the results of the study of the apoptotic proteins, we found an increased content of caspase-3 and the absence of significant changes in the content of the anti-apoptotic protein Bcl-2 in patients with lupus, which indicates active apoptotic activity. A comparative analysis of Beclin 1 and Vps34 showed their increased content in the cells of patients, which indicates the activation of autophagy. The analysis of two isoforms of LC3 protein revealed their low content in the group of patients. Since the scatter of indicators was very different from the average value, we analyzed these indicators depending on the severity of the disease. In the acute course group, high content of protein LC3-I was detected, the content of form II was lower. In the group with the subacute course, the number of both isoforms is lower than in the other groups. In the group with a chronic course, significant increases of protein LC3-II and a decrease in the ratio of LC3-I/LC3-II were found. Conclusion. The study showed that depending on the severity of systemic lupus erythematosus, the content of protein LC3 isoforms changes, which can be used for differential diagnosis of disease forms.

Download Full-text

Ameliorative Effects of Nano-Selenium Against Fluoride Stress Induced Hepatocytes Autophagy and Apoptosis in Mice

10.21203/rs.3.rs-44205/v1 ◽

2020 ◽

Author(s):

Yajing Wang ◽

Bingxian Liu ◽

Qingyue Han ◽

Khalid Mehmood ◽

Fazul Nabi ◽

...

Keyword(s):

Caspase 3 ◽

Beclin 1 ◽

Protective Effects ◽

Human Beings ◽

Caspase 9 ◽

Nano Material ◽

Cyt C ◽

Hepatocyte Damage

Abstract Background: Fluorine is widespread in the environment, and the injurious impacts of fluoride underscore its significance for public health. The long-term presence of fluorine in environment could be a risk in hepatotoxicity for both human beings and animals. Important role of selenium in mitigation of heavy metal toxicity via regulating autophagy and apoptosis is well-known. Further, nano-Se is a common artificial nano material, with higher biological activity and lower toxicity. The aim of the current study was to examine whether nano-Se supplementation can reduce the effects of fluoride-induced hepatocytes autophagy and apoptosis. Results: Here, we report that fluoride exposure induces apoptosis and autophagy with nucleus broken, dissolved and disappeared of hepatocyte, contributing to its hepatotoxicity. More importantly, Cyt-C and Beclin-1/Bcl-2 pathways are involved in the regulation of autophagy and apoptosis via targeting Caspase-9, Caspase-3, P53, Bax, LC3, ATG-5, P62 and mTOR expressions. Conclusion: Nano-Se is capable to alleviate fluoride-induced hepatocyte damage, that selenium can be prefer to prevent chronic fluorosis-induced autophagy and apoptosis by regulating Cyt-C and Beclin-1/Bcl-2 signaling pathway. In precisely, NaF-induced the liver injury by activating autophagy and apoptosis, which indicate that fluorine exposure, pose an ecological risk to human beings and animals. Nano-Se has protective effects against fluoride-induced hepatocytes.

Download Full-text

FAK and S6K1 Inhibitor, Neferine, Dually Induces Autophagy and Apoptosis in Human Neuroblastoma Cells

Molecules ◽

10.3390/molecules23123110 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3110 ◽

Cited By ~ 12

Author(s):

Dinh-Chuong Pham ◽

Yu-Chuan Chang ◽

Shian-Ren Lin ◽

Yuh-Ming Fuh ◽

May-Jywan Tsai ◽

...

Keyword(s):

Cell Migration ◽

Cancer Cells ◽

Molecular Mechanisms ◽

Caspase 3 ◽

Neuroblastoma Cells ◽

Beclin 1 ◽

Potential Candidate ◽

Human Neuroblastoma Cells ◽

Human Neuroblastoma ◽

Anti Cancer

Human neuroblastoma cancer is the most typical extracranial solid tumor. Yet, new remedial treatment therapies are demanded to overcome its sluggish survival rate. Neferine, isolated from the lotus embryos, inhibits the proliferation of various cancer cells. This study aimed to evaluate the anti-cancer activity of neferine in IMR32 human neuroblastoma cells and to expose the concealable molecular mechanisms. IMR32 cells were treated with different concentrations of neferine, followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to assess cell viability. In an effort to determine the molecular mechanisms in neferine-incubated IMR32 cells, cell cycle arrest, cell migration, and focal adhesion kinase (FAK), the 70-kDa ribosomal S6 kinase 1 (S6K1), poly (ADP-ribose) polymerase (PARP), caspase-3, Beclin-1, and microtubule-associated protein 1A/1B-light chain 3 (LC3) protein expressions were investigated. Neferine strongly disrupted the neuroblastoma cell growth via induction of G2/M phase arrest. Furthermore, neferine provoked autophagy and apoptosis in IMR32 cells, confirmed by p-FAK, and p-S6K1 reduction, LC3-II accumulation, Beclin-1 overexpression, and cleaved caspase-3/PARP improvement. Finally, neferine markedly retarded cell migration of neuroblastoma cancer cells. As a result, our findings for the first time showed an explicit anti-cancer effect of neferine in IMR32 cells, suggesting that neferine might be a potential candidate against human neuroblastoma cells to improve clinical outcomes with further in vivo investigation.

Download Full-text

Abstract 43: SIRT1 Enhances Oxidized Low-density Lipoprotein Induced Autophagy in Human Valve Interstitial Cells

Circulation Research ◽

10.1161/res.119.suppl_1.43 ◽

2016 ◽

Vol 119 (suppl_1) ◽

Author(s):

Huadong Li ◽

Jiawei Shi ◽

Weihua Qiao ◽

Nianguo Dong ◽

Gangjian Qin

Keyword(s):

Aortic Valve ◽

Heart Valve ◽

Western Blotting ◽

Caspase 3 ◽

Beclin 1 ◽

Calcium Deposition ◽

Control Group ◽

Ros Generation ◽

Aortic Valves ◽

Over Expression

Objectives: Sirt1, an NAD + -dependent class III deacetylase, has been shown to regulate autophagy during cardiac remodeling. Its role in calcific aortic valve disease has not been studied. Methods: Aortic valve samples were collected from patients with calcific aortic valve diseases complicated with hyper-lipidemia (at the time of heart valve replacement) (CAVD group), and from patients with dilated cardiomyopathy with non-calcified valves and a normal blood lipid profile (at the time of heart transplantation) (control group); the expression and distribution of calcification and Sirt1 in the aortic valves were analyzed by immuno-histochemical staining and Western blotting. In addition, primary human valvular intersititial cells (VICs) were isolated from normal valves in control group, and their response to DiI-ox-LDL treatments was analyzed in the presence of adenovirus-mediated Sirt1 over-expression (Ad-SIRT1) and Sirt1 inhibitor EX527, followed by assessments of the levels of reactive oxygen species (ROS), autophagy, apoptosis and calcification with biochemical methods. Results: The H&E, Masson, Von Kossa and α-SMA immuno-fluorescence staining showed that the aortic valves in CAVD group were thickened by excessive elastin, collagen expression and calcium deposition, which was associated with an elevated expression of osteocalcin and a reduced expression of Sirt1, as compared to control group. In vitro, the primary VICs were incubated with different concentrations (25, 50 and 100μg/mL) of ox-LDL for 48 h. ROS levels increased in an ox-LDL dose dependent manner. In addition, ox-LDL treatment led to a slight increase in the level of LC3 and Beclin-1, autophagic death-related hMOF and acetyl-histone H4, and a significant decrease in the level of Sirt1 through Western blotting. Interestingly, co-treatment of VICs with EX527 further increased ROS generation and expression of Caspase 3, Bax, and β-Catenin, while Sirt1 over-expression by Ad-SIRT1 resulted in increased expression of LC3 and Beclin-1, decreased expression of Caspase 3, Bax, Wnt3a and β-Catenin and reduced ROS generation. Conclusion: Our data suggest that Sirt1 may play a protective role in ox-LDL-induced VIC autophagy and heart valve calcification.

Download Full-text

Ischemic Preconditioning Promotes Autophagy and Alleviates Renal Ischemia/Reperfusion Injury

BioMed Research International ◽

10.1155/2018/8353987 ◽

2018 ◽

Vol 2018 ◽

pp. 1-11 ◽

Cited By ~ 8

Author(s):

Ying Xie ◽

Jing Xiao ◽

Chensheng Fu ◽

Zhenxing Zhang ◽

Zhibin Ye ◽

...

Keyword(s):

Ischemic Preconditioning ◽

Caspase 3 ◽

Cell Injury ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Kidney Injury ◽

Renal Ischemia ◽

Beclin 1 ◽

Autophagic Flux ◽

Dependent Manner

Autophagy is important for cellular survival during renal ischemia/reperfusion (I/R) injury. Ischemic preconditioning (IPC) has a strong renoprotective effect during renal I/R. Our study here aimed to explore the effect of IPC on autophagy during renal I/R injury. Rats were subjected to unilateral renal ischemia with or without prior IPC. Hypoxia/reoxygenation (H/R) injury was induced in HK-2 cells with or without prior hypoxic preconditioning (HPC). Autophagy and apoptosis were detected after reperfusion or reoxygenation for different time. The results showed that the levels of LC3II, Beclin-1, SQSTM1/p62, and cleaved caspase-3 were altered in a time-dependent manner during renal I/R. IPC further induced autophagy as indicated by increased levels of LC3II and Beclin-1, decreased level of SQSTM1/p62, and accumulation of autophagosomes compared to I/R groups at corresponding reperfusion time. In addition, IPC reduced the expression of cleaved caspase-3 and alleviated renal cell injury, as evaluated by the levels of serum creatinine (Scr), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) in renal tissues. In conclusion, autophagy and apoptosis are dynamically altered during renal I/R. IPC protects against renal I/R injury and upregulates autophagic flux, thus increasing the possibility for a novel therapy to alleviate I/R-induced acute kidney injury (AKI).

Download Full-text

Expression profiles of CD11b, galectin-1, beclin-1, and caspase-3 in nasal polyposis*

TURKISH JOURNAL OF MEDICAL SCIENCES ◽

10.3906/sag-1705-108 ◽

2017 ◽

Vol 47 ◽

pp. 1757-1764

Author(s):

Alper DİLCİ ◽

Nuray VAROL ◽

İlker KILIÇCIOĞLU ◽

Ece KONAC ◽

Utku AYDİL ◽

...

Keyword(s):

Nasal Polyposis ◽

Caspase 3 ◽

Expression Profiles ◽

Beclin 1

Download Full-text

The MicroRNA-210/Casp8ap2 Pathway Alleviates Hypoxia-Induced Injury in Myocardial Cells by Regulating Apoptosis and Autophagy

The Heart Surgery Forum ◽

10.1532/hsf.3173 ◽

2020 ◽

Vol 23 (6) ◽

pp. E797-E802

Author(s):

Kunsheng Li ◽

Jun Pan ◽

Qiuchang Li ◽

Shiliang Li ◽

Kai Li ◽

...

Keyword(s):

Myocardial Injury ◽

Caspase 3 ◽

Transfection Efficiency ◽

Beclin 1 ◽

Caspase 8 ◽

H9c2 Cells ◽

Myocardial Cells ◽

Western Blot Assay ◽

H9c2 Cardiomyocytes ◽

Associated Proteins

Aim: This study was conducted to investigate the role of the miR-210/Caspase8ap2 pathway in apoptosis and autophagy in hypoxic myocardial cells. Methods: The miR-control, miR-210 mimic, and miR-210 inhibitor were transfected into rat myocardial H9C2 cells. The transfection efficiency of exogenous miR-210 was determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). H9C2 cells were then treated with CoCl2 for 24, 48, and 72 h to generate a myocardial injury model. The apoptosis of H9C2 cells was assessed by flow cytometry. Additionally, a western blot assay was used to determine the expression of the autophagy-associated proteins light chain 3 (LC3), p62 and Beclin-1, and apoptosis-associated proteins Caspase8ap2, cleaved caspase 8, and cleaved caspase 3. Results: We determined that a 48 h hypoxia treatment duration in H9C2 cardiomyocytes induced myocardial injury. Additionally, the overexpression of miR-210 significantly inhibited cell apoptosis. MiR-210 suppressed autophagy by upregulating p62 and downregulating LC3II/I in hypoxic H9C2 cells. Caspase8ap2 was a putative target of miR-210, miR-210 mediated apoptosis, and autophagy of H9C2 cells via suppressing Caspase8ap2. Furthermore, the expression of caspase 8, caspase 3, and Beclin-1 were decreased in response to miR-210. Conclusion: miR-210 exhibits anti-apoptosis and anti-autophagy effects, which alleviate myocardial injury in response to hypoxia.

Download Full-text

Evaluation of programmed cell death processes on the lens epithelium of older dogs with diabetic and hypermature cataracts

Brazilian Journal of Veterinary Research and Animal Science ◽

10.11606/issn.1678-4456.bjvras.2018.133668 ◽

2018 ◽

Vol 55 (2) ◽

pp. e133668

Author(s):

Ana Paula Franco do Amaral Hvenegaard ◽

Paulo Sergio De Moraes Barros ◽

Angélica Mendonça Vaz Safatle ◽

Michelle Barbosa Pereira Braga-Sá ◽

Luana Vicente Melo ◽

...

Keyword(s):

Cell Death ◽

Cataract Surgery ◽

Trypan Blue ◽

Caspase 3 ◽

Posterior Capsule Opacification ◽

Beclin 1 ◽

Cellular Damage ◽

Lens Epithelial Cells ◽

Future Studies ◽

Proliferation And Differentiation

It is well known that posterior capsule opacification (PCO), one of the most common late postoperative complications of cataract surgery, is mainly caused by proliferation and differentiation of remaining lens epithelial cells (LECs) on the posterior lens capsule. Many authors suggest that alterations induced by the pathophysiology of cataracts, its metabolism and the use of 0.1% trypan blue (TB) must cause some degree of cellular damage on these cells, wicht would help to prevent and/or reduce the incidence of PCO after cataract surgery in humans. Therefore, the aim of this study was to evaluate the expression of cell death markers on LECs of older dogs with diabetic and hypermature cataracts, after capsulorhexis, both using 0.1% TB. Twenty samples collected from 13 dogs of different breeds, with ages varying from 8 to 12 years-old, with diabetic and hypermature cataracts, which had been subjected to phacoemulsification surgery (Phaco) using 0.1% TB for staining were studied. Animals were classified as dogs with diabetic (DC) and hypermature cataracts (HC), and expression of molecular markers for apoptosis and autophagy (caspase-3 and beclin-1) on LECs were obtained by immunofluorescence technique. The expression of caspase-3 and beclin-1 was observed in every studied sample and did not differ between groups. In conclusion, our findings suggest that apoptosis and autophagy processes occur to LECs in older dogs presenting diabetic and hypermature cataracts after Phaco utilizing 0.1% TB. Our results may be helpful to future studies of PCO in post-phacoemulsification surgery patients.

Download Full-text