Morphological and biochemical characterization and analysis of apoptosis

Allen, R. T.; Hunter, William J.; Agrawal, Devendra K.

doi:10.1016/s1056-8719(97)00033-6

Cited by 437 publications

(304 citation statements)

References 51 publications

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“…In order to further elucidate the role of leptin and gAcrp30 actions in the regulation of cytokine-and fatty acid-induced programmed cell death, we measured nucleosomal release as an early biochemical feature and quantitative marker of apoptosis [24]. Incubating INS-1 cells with the cytokine combination IL-1β/IFN-γ or palmitic acid resulted in a pronounced stimulation of DNA fragmentation (six-seven fold) (Fig.…”

Section: Resultsmentioning

confidence: 99%

Adiponectin counteracts cytokine- and fatty acid-induced apoptosis in the pancreatic beta-cell line INS-1

Rakatzi

Mueller

Ritzeler³

et al. 2004

Diabetologia

165

112

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Aims/hypothesis. Pancreatic beta-cell apoptosis is a common feature of Type 1 and Type 2 diabetes and leptin exerts an anti-apoptotic function in these cells. The beta-cell line INS-1 was used to test the hypothesis that the adipocyte hormone adiponectin might mediate an anti-apoptotic effect comparable to leptin. Methods. Apoptosis was induced by culturing cells with a cytokine combination (interleukin-1β/interferon-γ) or palmitic acid in absence or presence of leptin or the globular domain of adiponectin (gAcrp30), respectively.Results. INS-1 cells had a prominent sensitivity towards cytokine-and fatty acid-induced apoptosis, resulting in about three-and six-fold increases in caspase 3 activation and DNA fragmentation, respectively. gAcrp30 strongly (50-60%) inhibited palmitic acid-induced apoptosis, with a weaker effect against cytokine-induced apoptosis (35%). The same result was observed for leptin with both adipokines being non-additive. Reduction of apoptosis by an inhibitor of IκB-kinase (IKK) indicated the involvement of the nuclear factor (NF)-κB pathway in both cytokine-and fatty acid-induced apoptosis, however, leptin and gAcrp30 were unable to block NF-κB activation. Cytokine-and fatty-acid-induced suppression of glucose/forskolin-stimulated insulin secretion was completely prevented through the action of gAcrp30, whereas leptin was only effective against lipotoxicitymediated beta-cell dysfunction. Conclusion/interpretation. Our data show that gAcrp30 partially rescues beta cells from cytokineand fatty-acid-induced apoptosis and completely restores autoimmune-and lipotoxicity-induced dysfunction of insulin-producing cells. We suggest that gAcrp30 exerts its anti-apoptotic function without modulating NF-κB activation. This novel beta cell protective function of gAcrp30 might serve to counteract autoimmune-and lipotoxicity-induced beta-cell destruction. [Diabetologia (2004)

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Section: Resultsmentioning

confidence: 99%

Adiponectin counteracts cytokine- and fatty acid-induced apoptosis in the pancreatic beta-cell line INS-1

Rakatzi

Mueller

Ritzeler³

et al. 2004

Diabetologia

165

112

View full text Add to dashboard Cite

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“…They are primarily due to the loss of hair cells leading to degeneration of auditory neurons (Nordmann et al, 2000). Apoptosis, also known as programmed cell death, involves chromatin and cytoplasmic condensation, progressive contraction of cell volume, and preservation of the integrity of membrane-bound particles (Allen et al, 1997;Wyllie and Kerr 1980). Necrosis is generally characterized by early swelling of the cytoplasm and disintegration of cellular structures, which ultimately culminate in cell lysis and subsequent release of cellular debris into the extracellular space (Searle et al, 1982;Dive et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

A novel dual inhibitor of calpains and lipid peroxidation (BN82270) rescues the cochlea from sound trauma

et al. 2007

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Free radical and calcium buffering mechanisms are implicated in cochlear cell damage that has been induced by sound trauma. Thus in this study we evalua ted the therapeutic effect of a novel dual inhibitor of calpains and of lipid peroxidation (BN 82270) on the permanent hearing and hair cell loss induced by sound trauma.Perfusion of BN 82270 into the scala tympani of the guinea pig cochlea prevented the formation of calpain-cleaved fodrin, translocation of cytochrome c, DNA fragmentation and hair cell degeneration caused by sound trauma. This was confirmed by functional tests in vivo, showing a clear dose-dependent reduction of permanent hearing loss (ED 50 = 4.07 µM) with almost complete protection at 100 µM.Furthermore, BN82270 still remained effective even when applied onto the round window membrane after sound trauma had occurred, within a therapeutic window of 24hours. This indicates that BN 82270 may be of potential therapeutic value in treating the cochlea after sound trauma.

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“…Apoptosis study using the Annexin-V-FLUOS Staining kit demonstrated that SPME induced early apoptosis significantly 16 h post-treatment in both A549 and MCF-7 cells at 80 and 100 lg/ml, respectively. Internucleosomal DNA fragmentation is the primary hallmark to indicate an early event of apoptosis and it represents a point of no return from the path to cell death (Allen et al 1997). The observation from DAPI staining also supported that the above dose (80 and 100 lg/ml) of SPME is effective in inducing apoptosis in both types of cells.…”

Section: Discussionmentioning

confidence: 99%

In vitro anticancer activity of Spondias pinnata bark on human lung and breast carcinoma

et al. 2013

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Spondias pinnata, a commonly distributed tree in India, previously proven for various pharmacological properties and also reported for efficient anti-oxidant, free radical scavenging and iron chelating activity, continuing this, the present study is aimed to investigate the role of 70 % methanolic extract of S. pinnata bark (SPME) in promoting apoptosis in human lung adenocarcinoma cell line (A549) and human breast adenocarcinoma cell line (MCF-7). These two malignant cell lines and a normal cell line were treated with increasing concentrations of SPME and cell viability is calculated. SPME showed significant cytotoxicity to both A549 and MCF-7 cells with an IC 50 value of 147.84 ± 3.74 and 149.34 ± 13.30 lg/ml, respectively, whereas, comparatively no cytotoxicity was found in normal human lung fibroblast cell line (WI-38): IC 50 932.38 ± 84.44 lg/ ml. Flow cytometric analysis and confocal microscopic studies confirmed that SPME is able to induce apoptosis in both malignant cell lines. Furthermore, immunoblot result proposed the pathway of apoptosis induction by increasing Bax/Bcl-2 ratio in both cell types, which results in the activation of the caspasecascade and ultimately leads to the cleavage of Poly adeno ribose polymerase. For the first time this study proved the anticancer potential of SPME against human lung and breast cancer by inducing apoptosis through the modulation of Bcl-2 family proteins. This might take S. pinnata in light to investigate it for further development as therapeutic anticancer source.

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Morphological and biochemical characterization and analysis of apoptosis

Cited by 437 publications

References 51 publications

Adiponectin counteracts cytokine- and fatty acid-induced apoptosis in the pancreatic beta-cell line INS-1

Adiponectin counteracts cytokine- and fatty acid-induced apoptosis in the pancreatic beta-cell line INS-1

A novel dual inhibitor of calpains and lipid peroxidation (BN82270) rescues the cochlea from sound trauma

In vitro anticancer activity of Spondias pinnata bark on human lung and breast carcinoma

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