Caspase-independent Apoptosis Research Articles

Expression of Concern: The lysosome-associated apoptosis-inducing protein containing the pleckstrin homology (PH) and FYVE domains (LAPF), representative of a novel family of PH and FYVE domain-containing proteins, induces caspase-independent apoptosis via the lysosomal-mitochondrial pathway.

Expression of Concern: The lysosome-associated apoptosis-inducing protein containing the pleckstrin homology (PH) and FYVE domains (LAPF), representative of a novel family of PH and FYVE domain-containing proteins, induces caspase-independent apoptosis via the lysosomal-mitochondrial pathway.

О влиянии нового производного пиримидина PIR-4 на концентрацию маркеров внешнего и внутреннего путей апоптоза в условиях экспериментальной церебральной ишемии

Исследовано антиапоптотическое действие нового производного пиримидина под шифром PIR-4 в условиях экспериментальной фокальной церебральной ишемии у крыс линии Вистар, разделенных на 5 групп (n = 10). В первую группу включены ложнооперированные особи, вторая группа — крысы негативного контроля, получавшие взвесь очищенной воды и твина-80. Третья и четвертая группы представлены животными, получавшими препараты сравнения кавинтон (3,2 мг/кг) и глиатилин (60 мг/кг), соответственно. Пятой группе крыс вводили соединение PIR-4. Все соединения вводили сразу после моделирования ишемии и в течение 3 сут ежедневно. Модель фокальной ишемии мозга — окклюзия левой средней мозговой артерии (наркоз — хлоралгидрат, 350 мг/кг). В ходе проведенного исследования установлено, что на фоне моделируемой патологии возрастает концентрация фактора некроза опухоли (TNF-α), фактора, индуцирующего апоптоз (AIF), p53-зависимого модулятора апоптоза (PUMA), снижается уровень Jun-N-концевой киназы (JNK). Препараты сравнения кавинтон и глиатилин корректировали возникшие изменения, препятствуя развитию внешнего и внутреннего путей апоптоза. На фоне получения крысами вещества PIR-4 также уменьшал концентрацию маркеров каспаза-зависимого и каспаза-независимого путей апоптоза, при этом по антиапоптотическому эффекту, превосходя кавинтон и не уступая глиатилину.

Mitochondria-mediated Caspase-dependent and Caspase-independent apoptosis induced by aqueous extract from Moringa oleifera leaves in human melanoma cells.

Malignant melanoma is a very aggressive and serious type of cutaneous cancer. Previous studies indicated the anti-cancer activity of aqueous extract of Moringa oleifera Lam. leaves (MOE) against a variety of cell lines. However, there has not been much research about the effect of MOE on melanoma. Therefore, this study was about to investigate the anti-proliferation mediated by apoptosis of MOE on human melanoma cell lines. Furthermore, the related molecular mechanisms of the apoptosis were also examined. An aqueous extract of Moringa oleifera leaves was prepared and the anti-proliferative activity on melanoma cells and normal cells was tested using WST-1 assay. The apoptotic hallmarks including DNA condensation and phosphatidylserine (PS) externalization were assessed. The expression of apoptosis-related genes and the depolarization of mitochondrial membrane potential were then examined to clarify the underlying molecular mechanisms. MOE inhibited cell growth of A375 cells and A2058 cells in a dose-dependent manner but had little effect on human normal fibroblasts. The cell growth inhibition was induced by apoptosis which was expressed via chromatin condensation and PS externalization. MOE decreased mitochondrial membrane potential. Additionally, MOE increased Bax/Bcl-2 ratio, activated Caspase-3/7, Caspase-9, PARP and AIF translocation, leading to apoptotic cell death. Our study indicated that MOE exerted significant anti-cancer effects on melanoma cells in vitro which involved mitochondria-mediated Caspase-dependent and Caspase-independent apoptosis pathways. These results provided a scientific approach for using Moringa oleifera leaves as an alternative therapy to treat skin cancer.

Suppressing PARylation by 2',5'-oligoadenylate synthetase 1 inhibits DNA damage-induced cell death.

High expression of 2',5'-oligoadenylate synthetase 1 (OAS1), which adds AMP residues in 2',5' linkage to a variety of substrates, is observed in many cancers as a part of the interferon-related DNA damage resistance signature (IRDS). Poly(ADP-ribose) (PAR) is rapidly synthesized from NAD+ at sites of DNA damage to facilitate repair, but excessive PAR synthesis due to extensive DNA damage results in cell death by energy depletion and/or activation of PAR-dependent programmed cell death pathways. We find that OAS1 adds AMP residues in 2',5' linkage to PAR, inhibiting its synthesis invitro and reducing its accumulation in cells. Increased OAS1 expression substantially improves cell viability following DNA-damaging treatments that stimulate PAR synthesis during DNA repair. We conclude that high expression of OAS1 in cancer cells promotes their ability to survive DNA damage by attenuating PAR synthesis and thus preventing cell death.

Brain-dead donor heart conservation with a preservation solution supplemented by a conditioned medium from mesenchymal stem cells improves graft contractility after transplantation.

Hearts are usually procured from brain-dead (BD) donors. However, brain death may induce hemodynamic instability, which may contribute to posttransplant graft dysfunction. We hypothesized that BD-donor heart preservation with a conditioned medium (CM) from mesenchymal stem cells (MSCs) would improve graft function after transplantation. Additionally, we explored the PI3K pathway's potential role. Rat MSCs-derived CM was used for conservation purposes. Donor rats were either exposed to sham operation or brain death by inflation of a subdural balloon-catheter for 5.5hours. Then, the hearts were explanted, stored in cardioplegic solution-supplemented with either a medium vehicle (BD and sham), CM (BD+CM), or LY294002, an inhibitor of PI3K (BD+CM+LY), and finally transplanted. Systolic performance and relaxation parameters were significantly reduced in BD-donors compared to sham. After transplantation, systolic and diastolic functions were significantly decreased, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive cells and endonuclease G positive cells were increased in the BD-group compared to sham. Preservation of BD-donor hearts with CM resulted in a recovery of systolic graft function (dP/dtmax : BD+CM: 3148±178 vs BD: 2192±94mmHg/s at 110µL, P<.05) and reduced apoptosis. LY294002 partially lowered graft protection afforded by CM in the BD group. Our data suggest that PI3K/Akt pathway is not the primary mechanism of action of CM in improving posttransplant cardiac contractility and preventing caspase-independent apoptosis.

Inhibiting the interaction between apoptosis-inducing factor and cyclophilin A prevents brain injury in neonatal mice after hypoxia-ischemia

The interaction between apoptosis-inducing factor (AIF) and cyclophilin A (CypA) has been shown to contribute to caspase-independent apoptosis. Blocking the AIF/CypA interaction protects against glutamate-induced neuronal cell death in vitro, and the purpose of this study was to determine the in vivo effect of an AIF/CypA interaction blocking peptide (AIF(370-394)-TAT) on neonatal mouse brain injury after hypoxia-ischemia (HI). The pups were treated with AIF (370-394)-TAT peptide intranasally prior to HI. Brain injury was significantly reduced at 72 h after HI in the AIF(370-394)-TAT peptide treatment group compared to vehicle-only treatment for both the gray matter and the subcortical white matter, and the neuroprotection was more pronounced in males than in females. Neuronal cell death was evaluated in males at 8 h and 24 h post-HI, and it was decreased significantly in the CA1 region of the hippocampus and the nucleus habenularis region after AIF(370-394)-TAT treatment. Caspase-independent apoptosis was decreased in the cortex, striatum, and nucleus habenularis after AIF(370-394)-TAT treatment, but no significant change was found on caspase-dependent apoptosis as indicated by the number of active caspase-3-labeled cells. Further analysis showed that both AIF and CypA nuclear accumulation were decreased after treatment with the AIF(370-394)-TAT peptide. These results suggest that AIF(370-394)-TAT inhibited AIF/CypA translocation to the nucleus and reduced HI-induced caspase-independent apoptosis and brain injury in young male mice, suggesting that blocking AIF/CypA might be a potential therapeutic target for neonatal brain injury.

A novel sulfonamide derivative as a strong and selective apototic agent against hematological malignancies

Currently available chemotherapeutic drugs against hematological malignancies have several adverse effects and are associated with high mortality rates. Thus, in this study we evaluated the cytotoxic effect of 26 new sulfonamide derivatives on acute leukemia and multiple myeloma cells in order to try to discover a new selective and safe compound that might be used as a prototype for new chemotherapeutic agents. The most cytotoxic compound, DFS16, reduced the cell viability of K562, Jurkat and MM.1S cells in a concentration- and time- dependent manner and it was significantly less cytotoxic to non-tumor cells. On acute leukemia cells, sulfonamide DFS16 activated intrinsic and extrinsic apoptosis with Bax/Bcl-2 inversion, increased FasR expression and ΔΨm loss. In K562, DFS16 induced apoptosis by caspase-3 activation, while in Jurkat, it induced AIF release and caspase-3 independent apoptosis. In multiple myeloma, DFS16 induced cell cycle arrest at the G2/M phase and apoptosis with ΔΨm loss. Altogether, the results suggest that the new sulfonamide derivative DFS16 induces apoptotic-like cell death in acute leukemia and multiple myeloma cells. DFS16 is a promising new molecule that could be used as a prototype for the development of chemotherapeutics against hematological malignancies.

Antihelminthic Niclosamide Induces Autophagy and Delayed Apoptosis in Human Non-small Lung Cancer Cells In Vitro and In Vivo.

Niclosamide is an antihe-minthic drug that has shown cytotoxic effects on non-small cell lung carcinoma (NSCLC) cells. However, the exact mechanisms underlying the anti-tumour activity of niclosamide in NSCLC cancer cells remains to be defined. The aim of this study was to evaluate the antitumor activity of niclosamide in human A549 and CL1-5 non-small cell lung cancer cells using in vitro and in vivo. We investigated the effects of niclosamide on cell viability, apoptosis, the mitochondrial membrane potential (MMP; Δϕm), and autophagy and apoptosis-related protein expression in human A549 and CL1-5 non-small cell lung cancer cells. Niclosamide induced mainly caspase-independent apoptosis through apoptosis-inducible factor (AIF) translocation to the nucleus upon mitochondria damage. Moreover, niclosamide-induced autophagy may act as adaptive response against apoptosis. AMPK/AKT/mTOR pathway were involved in niclosamide-induced cell death and autophagy in response to ATP depletion. Furthermore, niclosamide efficiently suppressed tumor growth and induce autophagy in vivo. Niclosamide induced apoptosis by activating the intrinsic and caspase-independent pathway in human A549 and CL1-5 non-small cell lung cancer cells. Therefore, niclosamide is a potential candidate for anti-NSCLC therapy.

Progress of the anoikis effector bcl-2 inhibitor of transcription 1

bcl-2 inhibitor of transcription 1 (Bit1) is an anoikis effector that can play a role of caspase-independent apoptosis by down-regulating bcl-2 expression. Previous studies have found that Bit1 affects cell survival and apoptosis through Erk and FAK-PI3K-Akt-NF-κB pathways. It is worth noting that the expression of Bit1 has shown the obvious tumor specificity in different tumors, and it is closely related to TNM stage, differentiation and prognosis of the tumor. This review summarizes the expression, main mechanism, and significance of Bit1 in different tumors. Key words: Apoptosis; Neoplasms; Bcl-2 inhibitor of transcription 1

Sanggenol L promotes apoptotic cell death in melanoma skin cancer cells through activation of caspase cascades and apoptosis-inducing factor

Sanggenol L is one component of root bark of Morus alba. The molecular and cellular mechanisms of sanggenol L effects on melanoma cells are not well known. Recently, melanoma is the most common skin cancer with a high mortality rate not only in United States, but also in East Asia. Therefore, safe and effective treatments for melanoma treatment are required. In this study, we investigated whether or not sanggenol L possesses anti-cancer activity in human and mouse melanoma skin cancer cells. Sanggenol L treatment exerted significant cell growth inhibitory effects and inhibited colony formation capacity against B16, SK-MEL-2, and SK-MEL-28 melanoma skin cancer cells, whereas HaCaT human epithelial keratinocyte cells was unaffected by sanggenol L treatment. Sanggenol L treatment resulted in apoptotic cell death in melanoma skin cancer cells, which was characterized by accumulation of apoptotic cells, nuclear condensation, and apoptotic bodies. We also showed that sanggenol L treatment induced caspase-dependent apoptosis (up-regulation of Bax and cleaved-PARP or down-regulation of Bid, Bcl-2, procaspse-3, -8, and -9), induction of caspase-independent apoptosis (up-regulation of AIF and Endo G on cytosol) in melanoma skin cancer cells. These results suggest that sanggenol L induces caspase-dependent and -independent apoptosis in melanoma skin cancer cells.

Sanggenol L Induces Apoptosis and Cell Cycle Arrest via Activation of p53 and Suppression of PI3K/Akt/mTOR Signaling in Human Prostate Cancer Cells.

Prostate cancer is the most common cancer in Western countries. Recently, Asian countries are being affected by Western habits, which have had an important role in the rapid increase in cancer incidence. Sanggenol L (San L) is a natural flavonoid present in the root barks of Morus alba, which induces anti-cancer activities in ovarian cancer cells. However, the molecular and cellular mechanisms of the effects of sanggenol L on human prostate cancer cells have not been elucidated. In this study, we investigated whether sanggenol L exerts anti-cancer activity in human prostate cancer cells via apoptosis and cell cycle arrest. Sanggenol L induced caspase-dependent apoptosis (up-regulation of PARP and Bax or down-regulation of procaspase-3, -8, -9, Bid, and Bcl-2), induction of caspase-independent apoptosis (up-regulation of AIF and Endo G on cytosol), suppression of cell cycle (down-regulation of CDK1/2, CDK4, CDK6, cyclin D1, cyclin E, cyclin A, and cyclin B1 or up-regulation of p53 and p21), and inhibition of PI3K/Akt/mTOR signaling (down-regulation of PI3K, p-Akt, and p-mTOR) in prostate cancer cells. These results suggest the induction of apoptosis via suppression of PI3K/Akt/mTOR signaling and cell cycle arrest via activation of p53 in response to sanggenol L in prostate cancer cells.

Hericium erinaceus enhances neurotrophic factors and prevents cochlear cell apoptosis in senescence accelerated mice

• Hericium erinaceus prevents hearing degeneration in SAMP8 mice. • Hericium erinaceus increases nerve growth factor expression. • Hericium erinaceus prevents outer hair cell loss in the cochlea. • Hericium erinaceus reduces caspase-independent apoptosis. This study aimed to investigate the effects of Hericium erinaceus (HE) on auditory function. Twenty-four 9-month-old male senescence-accelerated prone 8 (SAMP8) mice were randomly and equally assigned to the control group (group A) and two groups supplemented with 215.25 (group B) or 430.5 mg/kg BW of HE (group C) for 12 weeks. Compared with group A, group C had significantly smaller threshold shifts by auditory brainstem responses at the end of study. HE increased nerve growth factor (NGF) expressions in the spiral ganglion neurons (SGNs) and the temporal lobes, and also prevented outer hair cells (OHCs) loss in the middle turn of cochlea. In addition, group C had significantly lower apoptosis-inducing factor and poly (ADP-ribose) polymerase-1 expressions in the middle turn of cochlea, brainstem and temporal lobes. In conclusions, HE could prevent hearing degeneration possibly by increasing neurotrophic expression, preventing OHCs loss, and reducing the caspase-independent apoptosis signalings.

Geranylated 4-phenylcoumarins extracted from Mesua elegans induced caspase-independent cell death in prostate cancer cell lines through calpain-2 and cathepsin B

Geranylated 4-phenylcoumarins DMDP-1 and DMDP-2 isolated from Mesua elegans were elucidated for their role in inducing caspase-independent programmed cell death (CI-PCD) in prostate cancer cell lines, PC-3 and DU 145, respectively. Cell homeostasis disruption was demonstrated upon treatment, as shown by the increase in calcium ion through colourimetric assay and endoplasmic reticulum (ER) stress markers GRP 78 and p-eIF2α through western blot. Subsequently, cytoplasmic death protease calpain-2 also showed increased activity during DMDP-1 & -2 treatments, while lysosomic death protease cathepsin B activity was significantly increased in PC-3 treated with DMDP-1. Flow cytometry showed a reduction in mitochondrial membrane potential in both cell lines, while western blotting showed translocation of mitochondrial death protease AIF into the cytoplasm in its truncated form. Furthermore, DMDP-1 & -2 treatments caused significant increase in superoxide level and oxidative DNA damage. Concurrent inhibition of calpain-2 and cathepsin B during the treatment showed an attenuation of cell death in both cell lines. Hence, DMDP-1 & -2 induce CI-PCD in prostate cancer cell lines through calpain-2 and cathepsin B.

ROS-Dependent Cell Death Induced by Parthenolide in Human Hepatoma Cell HepG2

With the increase of the action time and dosing concentration, the proliferation of HepG2 cell was inhibited and its vitality gradually decreased. FCM showed that, with the increase of PN action time, the MMP gradually decreased; calcium ions flowed inwards; the cell cycle was arrested in phase G1; the cell apoptosis rate, especially late apoptosis and necrotic cells, increased. The shear expression of apoptosis-related proteins caspase3 and caspase9 was up-regulated; the shear expression of AIF, MIF and PARP1 proteins associated with Caspase-independent apoptosis, i.e. Parthanatos apoptosis, showed time-dependent up-regulation; the long and short expressions of anti-apoptosis protein FLIP showed different degrees of decrease; the ex-pression of autophagy-related proteins LC3A/B and becin-1 was up-regulated; the expression of P62 protein was down-regulated; the expression of cycle- related proteins P53, P27 and P21 increased significantly; the expression of CyclinD1 and CyclinE1 decreased. FCM was used to detect the increase of ROS with the action time of PN; and its generation level showed an increasing trend; after the combination with ROS scavenger NAC, there was no significant difference in cell viability with the control group. There was no significant difference in the expression level of relevant cell death protein with DMSO control group. There was no difference in intracellular ROS generation level with the control group. Conclusion: PN induces the ROS generation in HepG2 cell, blocks its cycle and causes apoptosis and au-tophagy to play an anti-tumor effect.

Correlation between Antioxidant/Antimutagenic and Antiproliferative Activity of Some Phytochemicals.

Chemotherapeutic drugs have high toxicity associated with undesirable side-effects. Now, natural products are the most important anti-cancer agents because of their low toxicity and potential effectiveness. The half maximal inhibitory concentration (IC50) of amygdalin, naringenin and ellagic acid against breast, colon, and liver cell lines was estimated. The antimutagenic, free radical-, superoxide radical-, and hydroxyl radical- scavenging activities of these phytochemicals were measured. The expression of p53, bid, bax, bcl2, and caspases 9, 3, and 7 was measured by quantitative real-time polymerase chain reaction (qRT-PCR) in breast and liver cells. In addition, the active Caspase 3 protein was estimated in liver cells. Ellagic acid showed the highest antioxidant and antiproliferative activities. Amygdalin and naringenin with low and moderate antioxidant profiles showed a corresponding low and moderate cytotoxicity against cancer cell lines, respectively. Naringenin and ellagic acid had a significant antimutagenic activity which was detected by the Salmonella test. Ellagic acid offered a much better antimutagenic activity than naringenin. The apoptotic pathway evoked by ellagic acid in HepG2 and MCF-7 cells was investigated. The results showed that a caspase-dependent and a caspase-independent apoptosis occurred in MCF-7 and HepG2, respectively. The antimutagenic/antioxidant properties are well correlated with the antiproliferative activity of the phytochemicals investigated. This study proved that some easy, quick and cheap assays could predict the antiproliferative activity of many nutraceuticals. Finally, this platform could help in the discovery of new anticancer agents where hundreds of compounds are investigated in the pipeline of drug discovery.

Lupiwighteone induces caspase-dependent and -independent apoptosis on human breast cancer cells via inhibiting PI3K/Akt/mTOR pathway

Breast cancer is one of the most common causes of mortality in women. Lupiwighteone has anticancer effects in prostate cancer cells and neuroblastoma cells. However, the molecular and cellular mechanisms of lupiwighteone effects on human breast cancer cells are not as well known. In the present study, we investigated the effects of lupiwighteone on the proliferation and apoptosis of two different human cancer cells; MCF-7, an estrogen receptor (ER)-positive human breast cancer cell, and MDA-MB-231, a triple negative human breast cancer cell. Lupiwighteone treatment decreased the viability of MCF-7 and MDA-MB-231 cells. Lupiwighteone treatment resulted in apoptotic cell death in breast cancer cells, which was characterized by DNA fragmentation, accumulation of apoptotic cells, and nuclear condensation. We also showed that treatment with lupiwighteone induced caspase-dependent apoptosis (up-regulation of caspase-3, -7, -8, -9, PARP, and Bax or down-regulation of Bid, Bcl-2), induction of caspase-independent apoptosis (up-regulation of AIF and Endo G on cytosol), and inhibition of the PI3K/Akt/mTOR signaling pathway (down-regulation of PI3K, p-Akt, and p-mTOR) in both MCF-7 and MDA-MB-231 cells. These results suggest that lupiwighteone induces caspase-dependent and -independent apoptosis in both breast cancer cell lines via inhibiting PI3K/Akt/mTOR pathway.

A small cytotoxic peptide from frog elicits potent antitumor immunity to prevent local tumor growth and metastases.

Aim: Anticancer immunochemotherapy represents an attractive paradigm to improve therapeutic responses and reduce side effects. Results & methodology: Here, we show that a naturally occurring host defense peptide, HN-1 inhibited multiple malignant cells proliferation and tumor growth in a xenografted human breast tumor model. Acting through MAPK/NF-κB pathways, HN-1 induced a caspase-independent mitochondrial apoptosis, as indicated by a p53-dependent increase of Bax/Bcl-2 ratio and the nuclear translocation of apoptosis inducing factor. Besides, HN-1 augmented CD4+/CD8+ T cells in 4T1 mammary carcinoma model, by enhancing the serum levels of cancer immunity-associated effectors. Meanwhile, HN-1 decreased the angiogenesis and infiltration of the tumor-associated macrophages. Conclusion: HN-1 induces caspase-independent cancer cells apoptosis and boosts cancer-resolving immunity without inducing potentially harmful pro-inflammatory responses.

Carvacrol inhibits cadmium toxicity through combating against caspase dependent/independent apoptosis in PC12 cells

Carvacrol is a monoterpenic phenol found in essential oils, is considered a safe food additive, and possesses various therapeutic properties. Numerous studies have also deciphered the protective role of carvacrol on various cytotoxicities. We clarify the effects of carvacrol on cadmium-induced apoptosis in PC12 cells. Carvacrol while co-exposed with cadmium for 48 h raised PC12 cell viability in comparison to only cadmium exposed group. The co-exposure increased the cellular glutathione levels and promoted the expression of glutathione reductase. The magnitude of DNA fragmentation caused by cadmium was also ameliorated by carvacrol. Flow cytometry exhibited the apoptosis rate augmented by cadmium was reduced by carvacrol. Western blotting revealed that cadmium and carvacrol co-exposure alleviated the cadmium-induced down-regulations of mammalian target of rapamycin (mTOR), protein kinase B (Akt), nuclear factor kappa-light-chain-enhancer of activated B cells (NFКB), extracellular signal-regulated kinase-1 (ERK-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) expressions. The co-exposure also reversed action of cadmium by suppressing the cleavage of caspase 3 and reducing the cytosolic levels of cytochrome c and apoptosis inducing factor (AIF). Moreover, carvacrol upon co-exposure significantly increased the intracellular metallothionein content. In conclusion, carvacrol strongly reduced cadmium-triggered oxidative stress and caspase-dependent and caspase-independent apoptosis in PC12 cells.

Flow Cytometry Reveals the Nature of Oncotic Cells.

The term necrosis is commonly applied to cells that have died via a non-specific pathway or mechanism but strictly is the description of the degradation processes involved once the plasma membrane of the cell has lost integrity. The signalling pathways potentially involved in accidental cell death (ACD) or oncosis are under-studied. In this study, the flow cytometric analysis of the intracellular antigens involved in regulated cell death (RCD) revealed the phenotypic nature of cells undergoing oncosis or necrosis. Sodium azide induced oncosis but also classic apoptosis, which was blocked by zVAD (z-Vla-Ala-Asp(OMe)-fluoromethylketone). Oncotic cells were found to be viability+ve/caspase-3–ve/RIP3+ve/–ve (Receptor-interacting serine/threonine protein kinase 3). These two cell populations also displayed a DNA damage response (DDR) phenotype pH2AX+ve/PARP–ve, cleaved PARP induced caspase independent apoptosis H2AX–ve/PARP+ve and hyper-activation or parthanatos H2AX+ve/PARP+ve. Oncotic cells with phenotype cell viability+ve/RIP3–ve/caspase-3–ve showed increased DDR and parthanatos. Necrostatin-1 down-regulated DDR in oncotic cells and increased sodium azide induced apoptosis. This flow cytometric approach to cell death research highlights the link between ACD and the RCD processes of programmed apoptosis and necrosis.

Ethylmethylhydroxypyridine succinate, acetylcysteine and choline alphoscerate improve mitochondrial function under condition of cerebral ischemia in rat

The study was devoted to see the effect of ethylmethylhydroxypyridine succinate, acetylcysteine and choline alphoscerate on the mitochondrial function under condition of experimental cerebral ischemia in rat. An integrated approach was used with the assessment of changes in the respirometric and antiapoptotic functions of mitochondria. It was found that the application of the objects under study contributed to a decrease in the intensity of glycolysis and the restoration of aerobic glucose metabolism with an increase in ATP synthesis, as well as a decrease in the dissociation of reactions in mitochondrial complexes I, II, IV and V. In addition, the introduction of the studied drugs reduced the intensity of caspase-dependent and caspase-independent apoptosis reactions, which, together with the restoration of mitochondrial respiratory function, contributed to the decrease in the size of the necrosis zone of the brain.&#x0D; Video Clip of Methodology:&#x0D; Rat middle cerebral artery occlusion: 8 min 7 sec Full Screen Alternate