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pCramoll and rCramoll as New Preventive Agents against the Oxidative Dysfunction Induced by Hydrogen Peroxide.

da Silva LC, Alves NM, de Castro MC, Higino TM, da Cunha CR, Pereira VR, da Paz NV, Coelho LC, Correia MT, de Figueiredo RC - Oxid Med Cell Longev (2015)

Bottom Line: Oxidative stress plays an important role in the induction of cell death and is associated with various pathologic disorders; therefore, the search for natural products that attenuate the effects produced by oxidant agents is greatly increased.The maximum protective effects were 96.85 ± 15.59% (rCramoll) and 59.48 ± 23.44% (pCramoll).The deleterious effects of H2O2 on cell proliferation were reduced to 10.83% (pCramoll) and 24.17% (rCramoll).

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Bioquímica de Proteínas, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Avenida Professor Moraes Rêgo, 1235 Cidade Universitária, 50670-901 Recife, PE, Brazil ; Functional Genomics, Department of Biology, Faculty of Science, University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen, Denmark.

ABSTRACT
Oxidative stress plays an important role in the induction of cell death and is associated with various pathologic disorders; therefore, the search for natural products that attenuate the effects produced by oxidant agents is greatly increased. Here, the protective effects of native lectin from Cratylia mollis seeds (pCramoll) and recombinant Cramoll 1 (rCramoll) against H2O2-induced oxidative stress in Vero cells were evaluated. Both lectins significantly attenuated the H2O2-induced cytotoxicity in a concentration-dependent way. The maximum protective effects were 96.85 ± 15.59% (rCramoll) and 59.48 ± 23.44% (pCramoll). The Live/Dead analysis showed a reduction in the percentage of dead cells from 65.04 ± 3.29% (H2O2) to 39.77 ± 2.93% (pCramoll) and 13.90 ± 9.01% (rCramoll). The deleterious effects of H2O2 on cell proliferation were reduced to 10.83% (pCramoll) and 24.17% (rCramoll). Lectins treatment attenuated the excessive superoxide production, the collapse of the mitochondrial membrane potential, and the lysosomal and DNA damage in H2O2-treated cells. In conclusion, our results suggest that pCramoll and rCramoll blocked H2O2-induced cytotoxicity through decreasing reactive oxygen species, restoring the mitochondrial potential, preventing the lysosomal damage and DNA fragmentation, and thus promoting cell survival and proliferation.

No MeSH data available.


Related in: MedlinePlus

Effect of pCramoll and rCramoll on the DNA damage induced by H2O2 in Vero cells by TUNEL analysis using flow cytometry. (∗) Significant differences in relation to H2O2.
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fig7: Effect of pCramoll and rCramoll on the DNA damage induced by H2O2 in Vero cells by TUNEL analysis using flow cytometry. (∗) Significant differences in relation to H2O2.

Mentions: The formation of DNA ladders is a consequence of a specific nucleosomal-sized fragmentation and is a conventional event in apoptotic process. The TUNEL assay was carried out to detect the extension of DNA degradation in apoptotic cells. DNA damage was quantified in relation to control cells. As expected, no significant DNA injury could be detected in the control cells. On the other hand, H2O2-treated cells presented an increase in the TUNEL fluorescence intensity, indicative of intense DNA fragmentation (this fluorescence was 170.41% higher than control cell). Pretreatment with both lectins leads to decrease in TUNEL staining in about 2-fold (61.62 ± 1.44% for pCramoll and 55.53 ± 5.86% for rCramoll) (Figure 7).


pCramoll and rCramoll as New Preventive Agents against the Oxidative Dysfunction Induced by Hydrogen Peroxide.

da Silva LC, Alves NM, de Castro MC, Higino TM, da Cunha CR, Pereira VR, da Paz NV, Coelho LC, Correia MT, de Figueiredo RC - Oxid Med Cell Longev (2015)

Effect of pCramoll and rCramoll on the DNA damage induced by H2O2 in Vero cells by TUNEL analysis using flow cytometry. (∗) Significant differences in relation to H2O2.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4632182&req=5

fig7: Effect of pCramoll and rCramoll on the DNA damage induced by H2O2 in Vero cells by TUNEL analysis using flow cytometry. (∗) Significant differences in relation to H2O2.
Mentions: The formation of DNA ladders is a consequence of a specific nucleosomal-sized fragmentation and is a conventional event in apoptotic process. The TUNEL assay was carried out to detect the extension of DNA degradation in apoptotic cells. DNA damage was quantified in relation to control cells. As expected, no significant DNA injury could be detected in the control cells. On the other hand, H2O2-treated cells presented an increase in the TUNEL fluorescence intensity, indicative of intense DNA fragmentation (this fluorescence was 170.41% higher than control cell). Pretreatment with both lectins leads to decrease in TUNEL staining in about 2-fold (61.62 ± 1.44% for pCramoll and 55.53 ± 5.86% for rCramoll) (Figure 7).

Bottom Line: Oxidative stress plays an important role in the induction of cell death and is associated with various pathologic disorders; therefore, the search for natural products that attenuate the effects produced by oxidant agents is greatly increased.The maximum protective effects were 96.85 ± 15.59% (rCramoll) and 59.48 ± 23.44% (pCramoll).The deleterious effects of H2O2 on cell proliferation were reduced to 10.83% (pCramoll) and 24.17% (rCramoll).

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Bioquímica de Proteínas, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Avenida Professor Moraes Rêgo, 1235 Cidade Universitária, 50670-901 Recife, PE, Brazil ; Functional Genomics, Department of Biology, Faculty of Science, University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen, Denmark.

ABSTRACT
Oxidative stress plays an important role in the induction of cell death and is associated with various pathologic disorders; therefore, the search for natural products that attenuate the effects produced by oxidant agents is greatly increased. Here, the protective effects of native lectin from Cratylia mollis seeds (pCramoll) and recombinant Cramoll 1 (rCramoll) against H2O2-induced oxidative stress in Vero cells were evaluated. Both lectins significantly attenuated the H2O2-induced cytotoxicity in a concentration-dependent way. The maximum protective effects were 96.85 ± 15.59% (rCramoll) and 59.48 ± 23.44% (pCramoll). The Live/Dead analysis showed a reduction in the percentage of dead cells from 65.04 ± 3.29% (H2O2) to 39.77 ± 2.93% (pCramoll) and 13.90 ± 9.01% (rCramoll). The deleterious effects of H2O2 on cell proliferation were reduced to 10.83% (pCramoll) and 24.17% (rCramoll). Lectins treatment attenuated the excessive superoxide production, the collapse of the mitochondrial membrane potential, and the lysosomal and DNA damage in H2O2-treated cells. In conclusion, our results suggest that pCramoll and rCramoll blocked H2O2-induced cytotoxicity through decreasing reactive oxygen species, restoring the mitochondrial potential, preventing the lysosomal damage and DNA fragmentation, and thus promoting cell survival and proliferation.

No MeSH data available.


Related in: MedlinePlus