Limits...
Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver.

Srilaxmi P, Sareddy GR, Kavi Kishor PB, Setty OH, Babu PP - BMC Pharmacol. (2010)

Bottom Line: Natansnin treatment significantly decreased the levels of CCl4 induced apoptotic proteins and inflammatory mediators.Further natansinin treatment significantly inhibited the CCl4 induced apoptosis which was evident form the reduced TUNEL positive cells.This protective effect of natansnin can be correlated to its direct antioxidant effect.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics, Osmania University, Hyderabad, India.

ABSTRACT

Background: Carbon tetra chloride (CCl4), an industrial solvent, is a hepatotoxic agent and it is the well established animal model for free radical-induced liver injury. The present investigation was carried out to establish the protective effect of natansnin, a novel dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver.

Results: CCl4 significantly increased the levels of lipid peroxides, oxidized glutathione and decreased the levels of reduced glutathione, SOD and CAT. CCl4 induce marked histopathological changes and increase in the levels of apoptotic proteins. CCl4 treatment significantly increased the levels of apoptotic proteins such as caspases-3, PARP, Bax, Bid and cytochrome C and also increased the levels of inflammatory mediators iNos and Cox-2. Natansnin treatment significantly decreased the levels of CCl4 induced apoptotic proteins and inflammatory mediators. Further natansinin treatment significantly inhibited the CCl4 induced apoptosis which was evident form the reduced TUNEL positive cells.

Conclusions: In conclusion, our study demonstrated the protective effect of natansnin against CCl4 induced oxidative stress and cellular degeneration in rat liver tissue. This protective effect of natansnin can be correlated to its direct antioxidant effect.

Show MeSH

Related in: MedlinePlus

Effect of CCl4 with or without prior administration of natansnin on Bax and Bid. Mitochondrial protein extracted from control, CCl4 and natansnin (10 mg/kg, 20 mg/kg body weight) treated rats and separated on 10% SDS gels and transferred on to nitrocellulose membranes. Immunoblots were detected with Bax and Bid specific primary antibodies. The expression levels of β-actin were used as loading controls. Densitometric analysis showing the protein levels of Bax and Bid were increased in CCl4 group while treatment with natansnin significantly decreased the protein levels. Data are represented as mean ± standard deviation from three independent experiments (*p < 0.05 indicate significant difference relative to the CCl4 treated rats).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2967507&req=5

Figure 9: Effect of CCl4 with or without prior administration of natansnin on Bax and Bid. Mitochondrial protein extracted from control, CCl4 and natansnin (10 mg/kg, 20 mg/kg body weight) treated rats and separated on 10% SDS gels and transferred on to nitrocellulose membranes. Immunoblots were detected with Bax and Bid specific primary antibodies. The expression levels of β-actin were used as loading controls. Densitometric analysis showing the protein levels of Bax and Bid were increased in CCl4 group while treatment with natansnin significantly decreased the protein levels. Data are represented as mean ± standard deviation from three independent experiments (*p < 0.05 indicate significant difference relative to the CCl4 treated rats).

Mentions: Immunoblot analysis of caspase-3, PARP, cytochrome C, iNOS and COX-2 were performed in whole cell lysates of liver samples of all the animals. Also, immunoblot analysis of Bax and Bid were performed in mitochondrial fractions of all liver samples. Mitochondrial cytochrome C, detected as a single band of molecular mass (14 kDa) was increased in the whole cell lysate of CCl4 treated rats, compared to controls (Figure 7). Densitometric analysis of this protein is shown in Figure 7. Release of cytochrome C from mitochondria to cytosol is clearly noticed. Immuno blot analysis of pro caspase-3 (17 kDa) and active caspase-3 (32 kDa) increased in CCl4 treated rats and densitometric analysis of this protein is also shown in Figure 7. Immuno blot analysis of cleaved PARP fragments (37 kDa, 51 kDa, 64 kDa, 89 kDa and 98 kDa) increased in CCl4 treated rats and also its densitometric analysis is shown in Figure 8. The cleavage of PARP was clearly observed. The levels of these proteins were appreciably reduced in natansnin treated, compared to CCl4 exposed animals indicating a decrease in the cell death-signaling pathway. Immuno blot analysis of apoptotic proteins like Bax (21kDa) and Bid (23kDa) in mitochondrial fractions of CCl4 treated rats revealed an increase and are shown in Figure 9. The immune blot analysis of inflammatory proteins iNOS (130 kDa) and COX-2 (74 kDa) considerably increased in CCl4 treated rats and are shown in Figure 10. The densitometric analyses of these proteins are also shown in Figure 10. Natansnin treatment reduced the level of these inflammatory proteins. However, levels of these proteins reduced significantly in natansnin (both concentrations) treated rats, compared to CCl4 exposed ones indicating a decrease in the cell death.


Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver.

Srilaxmi P, Sareddy GR, Kavi Kishor PB, Setty OH, Babu PP - BMC Pharmacol. (2010)

Effect of CCl4 with or without prior administration of natansnin on Bax and Bid. Mitochondrial protein extracted from control, CCl4 and natansnin (10 mg/kg, 20 mg/kg body weight) treated rats and separated on 10% SDS gels and transferred on to nitrocellulose membranes. Immunoblots were detected with Bax and Bid specific primary antibodies. The expression levels of β-actin were used as loading controls. Densitometric analysis showing the protein levels of Bax and Bid were increased in CCl4 group while treatment with natansnin significantly decreased the protein levels. Data are represented as mean ± standard deviation from three independent experiments (*p < 0.05 indicate significant difference relative to the CCl4 treated rats).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2967507&req=5

Figure 9: Effect of CCl4 with or without prior administration of natansnin on Bax and Bid. Mitochondrial protein extracted from control, CCl4 and natansnin (10 mg/kg, 20 mg/kg body weight) treated rats and separated on 10% SDS gels and transferred on to nitrocellulose membranes. Immunoblots were detected with Bax and Bid specific primary antibodies. The expression levels of β-actin were used as loading controls. Densitometric analysis showing the protein levels of Bax and Bid were increased in CCl4 group while treatment with natansnin significantly decreased the protein levels. Data are represented as mean ± standard deviation from three independent experiments (*p < 0.05 indicate significant difference relative to the CCl4 treated rats).
Mentions: Immunoblot analysis of caspase-3, PARP, cytochrome C, iNOS and COX-2 were performed in whole cell lysates of liver samples of all the animals. Also, immunoblot analysis of Bax and Bid were performed in mitochondrial fractions of all liver samples. Mitochondrial cytochrome C, detected as a single band of molecular mass (14 kDa) was increased in the whole cell lysate of CCl4 treated rats, compared to controls (Figure 7). Densitometric analysis of this protein is shown in Figure 7. Release of cytochrome C from mitochondria to cytosol is clearly noticed. Immuno blot analysis of pro caspase-3 (17 kDa) and active caspase-3 (32 kDa) increased in CCl4 treated rats and densitometric analysis of this protein is also shown in Figure 7. Immuno blot analysis of cleaved PARP fragments (37 kDa, 51 kDa, 64 kDa, 89 kDa and 98 kDa) increased in CCl4 treated rats and also its densitometric analysis is shown in Figure 8. The cleavage of PARP was clearly observed. The levels of these proteins were appreciably reduced in natansnin treated, compared to CCl4 exposed animals indicating a decrease in the cell death-signaling pathway. Immuno blot analysis of apoptotic proteins like Bax (21kDa) and Bid (23kDa) in mitochondrial fractions of CCl4 treated rats revealed an increase and are shown in Figure 9. The immune blot analysis of inflammatory proteins iNOS (130 kDa) and COX-2 (74 kDa) considerably increased in CCl4 treated rats and are shown in Figure 10. The densitometric analyses of these proteins are also shown in Figure 10. Natansnin treatment reduced the level of these inflammatory proteins. However, levels of these proteins reduced significantly in natansnin (both concentrations) treated rats, compared to CCl4 exposed ones indicating a decrease in the cell death.

Bottom Line: Natansnin treatment significantly decreased the levels of CCl4 induced apoptotic proteins and inflammatory mediators.Further natansinin treatment significantly inhibited the CCl4 induced apoptosis which was evident form the reduced TUNEL positive cells.This protective effect of natansnin can be correlated to its direct antioxidant effect.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics, Osmania University, Hyderabad, India.

ABSTRACT

Background: Carbon tetra chloride (CCl4), an industrial solvent, is a hepatotoxic agent and it is the well established animal model for free radical-induced liver injury. The present investigation was carried out to establish the protective effect of natansnin, a novel dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver.

Results: CCl4 significantly increased the levels of lipid peroxides, oxidized glutathione and decreased the levels of reduced glutathione, SOD and CAT. CCl4 induce marked histopathological changes and increase in the levels of apoptotic proteins. CCl4 treatment significantly increased the levels of apoptotic proteins such as caspases-3, PARP, Bax, Bid and cytochrome C and also increased the levels of inflammatory mediators iNos and Cox-2. Natansnin treatment significantly decreased the levels of CCl4 induced apoptotic proteins and inflammatory mediators. Further natansinin treatment significantly inhibited the CCl4 induced apoptosis which was evident form the reduced TUNEL positive cells.

Conclusions: In conclusion, our study demonstrated the protective effect of natansnin against CCl4 induced oxidative stress and cellular degeneration in rat liver tissue. This protective effect of natansnin can be correlated to its direct antioxidant effect.

Show MeSH
Related in: MedlinePlus