Detecting Role of Apoptosis in Mediating Cyclophosphamide Induced Teratogenesis In Vitro

Singh, Gyanendra; Sinha, Neeraj; Koushik, C Jayendra; Mathur, Suvigya; Srivastava, Sudhir

doi:10.1080/15376520500194791

Cited by 6 publications

(10 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, at lower doses, the effects were not so significant as compared to untreated control. Whereas at high drug concentrations, a meager increase in necrosis was also observed, which might be because of cytotoxic nature of the drug at these high concentrations as already documented by several studies [34][35][36]. In addition, the cell cycle analysis also showed a sub-Go peak of apoptotic cells, which increased with increasing drug concentration.…”

Section: Discussionsupporting

confidence: 62%

“…However, their number at 10 μg/mL culture (LD) was very low with respect to untreated (control) and was not found significant ( P > 0.05). As discussed by several experts, a sub‐Go peak can be another qualitative indicator of apoptosis [34–36,42,43]. Hence, we employ sub‐Go peak, as an additional confirmatory indicator of apoptosis.…”

Section: Resultsmentioning

confidence: 99%

“…At higher concentration, excessive cell death in defined regions of the embryo such as lesions in forebrain and in fore and hind limbs may lead to characteristic malformations, which are observed in the form of necrotic cell death concurrently with apoptosis. DNA fragmentation has been extensively reported as a biochemical hallmark of apoptosis as described previously [34–36]. During apoptosis, DNA fragmentation is catalyzed by Ca 2+ ‐ and Mg 2+ ‐activated endonuclease and is characterized by double‐strand cleavage at the linker regions between nucleosomes resulting in the production of DNA fragments, which are multiples of approximately 180 bp [37,38].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Studying significance of apoptosis in mediating tolbutamide‐induced teratogenesis in vitro

Singh

Kumar

Sinha³

2011

Fundamemntal Clinical Pharma

Self Cite

View full text Add to dashboard Cite

The incidence of type 2 diabetes mellitus (non-insulin-dependent diabetes mellitus) is growing worldwide and poses a serious public health problem in a current paradigm of changing life style and food habits. Tolbutamide (sulfonylurea) is among the commonly used anti-diabetic drugs worldwide for treating type 2 diabetes and is known to cause congenital malformations in animals. In this study, the effect of tolbutamide on major organogenesis period and the possible involvement of apoptosis in mediating congenital malformations have been carried out. In the present study design, post-implantation rat embryos of day 11 were cultured for 24 h with various concentrations of tolbutamide, i.e., 10, 100, and 1000 μg/mL cultures, respectively. The growth and developmental of each embryo was evaluated and compared with control ones for the presence of any malformations. The tolbutamide decreased all growth and developmental parameters in a concentration-dependent manner, when compared with control. However, exposure to tolbutamide at 10 μg/mL culture did not show any significant effect on embryonic growth and development in vitro. In parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were carried out followed by quantitation by 3'-OH labeling of cultured rat embryos to examine the role of apoptosis in bringing about tolbutamide-induced teratogenesis. All results were found to be dose dependent and an increase in apoptosis in embryonic tissues may be related to the increased risk of congenital malformations. The outcome of the research suggested that apoptosis might be involved in mediating teratogenesis of tolbutamide in vitro. Further research is warranted to fully understand this mechanism.

show abstract

Section: Discussionsupporting

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Studying significance of apoptosis in mediating tolbutamide‐induced teratogenesis in vitro

Singh

Kumar

Sinha³

2011

Fundamemntal Clinical Pharma

Self Cite

View full text Add to dashboard Cite

show abstract

“…Many studies detected increased apoptosis after exposure to various teratogens suggesting a correlation between increased apoptosis and teratogen‐induced fetal malformations (Torchinsky et al., ; Savion et al., ; Singh et al., ). Increased apoptosis was observed in developing embryos exposed to cyclophosphamide (Chen et al., ; Torchinsky et al., ; Savion et al., ; Singh et al., ), arsenic (Li et al., ), salicylic acid (Singh et al., ), homocysteine (Li et al., ) and these studies proposed a relation between increased apoptosis and teratogenesis. Exposure to retinoic acid (Sarkar and Sharma, ) and nicotine (Zhao et al., ) resulted in apoptosis in the neuroepithelium of developing mouse embryos suggesting apoptosis as a possible mechanism for teratogen‐induced NTDs.…”

Section: Discussionmentioning

confidence: 99%

Reduction in Valproic Acid–Induced Neural Tube Defects by Maternal Immune Stimulation: Role of Apoptosis

Mallela

Hrubec

2012

Birth Defects Research Pt B

View full text Add to dashboard Cite

Teratogenic deregulation of apoptosis during development is a possible mechanism for birth defects. Administration of valproic acid (VA) during first trimester of pregnancy causes neural tube defects (NTDs). Nonspecific stimulation of the mother’s immune system has been shown to reduce various teratogen-induced fetal malformations including NTDs in rodents. This present study investigated the role of reduced apoptosis by maternal immune stimulation in prevention of VA-induced NTDs in CD-1 mice. Prevention of VA-induced NTDs by nonspecific maternal immune stimulation using IFNγ was employed to evaluate the role of reduced apoptosis by IFNγ in this protective mechanism. Apoptosis was quantified using flow cytometry. Terminal Transferase dUTP Nick End Labeling assay was used to localize the apoptosis. Increased apoptosis, suggesting involvement in VA teratogenicity, was observed along the neural tube in both normal and abnormal embryos from VA-exposed dams. Increased apoptosis in normal VA-exposed embryos suggests that VA may alter other cellular processes such as cell proliferation and differentiation in addition to apoptosis. Apoptotic levels in embryos with closed neural tubes from IFNγ + VA dams were similar to controls indicating resistance to VA-induced apoptosis and protection against teratogenicity of VA. In IFNγ + VA exposed embryos with open neural tubes, maternal immune stimulation failed to regulate apoptosis resulting in an NTD. Overall, these results suggest that VA alters several biological processes including apoptosis in the developing embryos to induce fetal malformations. Resistance to VA-induced apoptosis in embryos resulting from maternal immune stimulation may be involved in protective mechanism.

show abstract

“…However, their number at LD (3.75 mg/ml)-culture was not found to be significant with respect to untreated (control; p > 0.05). As discussed in earlier studies, a sub-G0 peak could be another qualitative parameter of apoptosis (Darzynkiewicz et al, 1997;Del Bino et al, 1999;Singh and Sinha, 2010;Singh et al, 2005Singh et al, , 2009Singh et al, , 2011. Hence, the sub-G0 peak was employed as an additional confirmatory indicator of apoptosis.…”

Section: Sub-g0 Assaymentioning

confidence: 98%

Role of apoptosis in mediating diclofenac-induced teratogenesis

et al. 2013

Self Cite

View full text Add to dashboard Cite

Diclofenac (DCF) is among the most commonly used nonsteroidal anti-inflammatory drugs worldwide for the treatment of various conditions in postpubertal women. However, very limited information is available regarding its safety during pregnancy and teratogenecity. The present study was designed to elucidate the effects of DCF on the developing rat embryos during the major organogenesis period and investigate the critical role of apoptosis in bringing about these congenital anomalies. Embryos were exposed in vitro to various concentrations of DCF, that is, 0, 3.75, 7.5 and 15 µg/ml for 24 h, respectively, and examined for the growth and differentiation at the end of the culture period for the presence of any specific malformations. Growth and developmental parameters such as weight of embryos, crown-rump length and number of somites were found to be lower in the embryos exposed to high concentrations of DCF (7.5 and 15.0 μg/ml) when compared with the untreated control. However, no significant difference in growth parameters was found between embryos exposed to 3.75 µg/ml and the control group. In parallel to this, flow cytometric analysis and DNA quantitation of cultured rat embryos were performed to verify the involvement of apoptosis in mediating DCF-induced teratogenesis. A concentration-dependent increase in apoptosis in embryos suggests a possible engagement of apoptosis in the role of DCF as a teratogenic agent. A detailed analysis of the actual effect of DCF on cellular apoptotic machinery necessitates further evaluation.

show abstract

Detecting Role of Apoptosis in Mediating Cyclophosphamide Induced Teratogenesis In Vitro

Cited by 6 publications

References 19 publications

Studying significance of apoptosis in mediating tolbutamide‐induced teratogenesis in vitro

Studying significance of apoptosis in mediating tolbutamide‐induced teratogenesis in vitro

Reduction in Valproic Acid–Induced Neural Tube Defects by Maternal Immune Stimulation: Role of Apoptosis

Role of apoptosis in mediating diclofenac-induced teratogenesis

Contact Info

Product

Resources

About