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Antioxidant and prophylactic effects of Delonix elata L., stem bark extracts, and flavonoid isolated quercetin against carbon tetrachloride-induced hepatotoxicity in rats.

Krishnappa P, Venkatarangaiah K - Biomed Res Int (2014)

Bottom Line: Bioassay-guided fractionation of DSE has resulted in the isolation and characterization of quercetin.DSE and quercetin have shown significant prophylactic effects by restoring the liver function markers (AST, ALT, ALP, serum bilirubin, and total protein) and antioxidant enzymes (SOD, CAT, GPx, and GST).These results were proved to be hepatoprotective at par with silymarin and well supported by the histological observations of liver sections with distinct hepatic cells, and mild degree of fatty change and necrosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Post Graduate Studies and Research in Biotechnology, Kuvempu University, Shankaraghatta, Karnataka 577 451, India.

ABSTRACT
Delonix elata L. (Ceasalpinaceae), is widely used by the traditional medical practitioners of Karnataka, India, to cure jaundice, and bronchial and rheumatic problems. The objective of this study was to screen the in vitro antioxidant and hepatoprotective activity of the stem bark extracts against CCl4-induced liver damage in rats. Among different stem bark extracts tested, the ethanol extract (DSE) has shown significant in vitro antioxidant property in radicals scavenging, metal chelating, and lipid peroxidation inhibition assays. HPLC analysis of the DSE revealed the presence of known antioxidant molecules, namely, gallic acid, ellagic acid, coumaric acid, quercetin, and rutin. Bioassay-guided fractionation of DSE has resulted in the isolation and characterization of quercetin. DSE and quercetin have shown significant prophylactic effects by restoring the liver function markers (AST, ALT, ALP, serum bilirubin, and total protein) and antioxidant enzymes (SOD, CAT, GPx, and GST). These results were proved to be hepatoprotective at par with silymarin and well supported by the histological observations of liver sections with distinct hepatic cells, and mild degree of fatty change and necrosis. The results indicated that the DSE and quercetin were significant for prophylactic activity against CCl4-induced liver damage in rats. This activity could be attributed to the antioxidant constituents in the DSE and hence justified the ethnomedicinal claims.

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Histopathology of liver tissues. (a) Liver section of normal control rats showing normal architecture; (b) Liver section of CCl4 treated rats showing massive fatty changes, cellular necrosis, vacuolization, and ballooning degeneration; (c) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSC showing mild fatty vacuolation and cellular necrosis; (d) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSC showing cellular regeneration, mild degree of cell necrosis, and fatty vacuolation; (e) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSE showing few fatty vacuoles, restoration of hepatocytes, and lesser necrosis; (f) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSE showing minimal inflammatory cellular infiltration and almost near normal liver architecture; (g) Liver section of rats treated with CCl4 and quercetin showing recovery from the toxic effect with normal arrangement of hepatocytes with little evidence of fatty vacuoles and cellular necrosis (h) Liver histology of animals treated with silymarin showing normal histology with least parenchymal injury. (1-cellular necrosis; 2-vacuolization; 3-ballooning degeneration).
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fig5: Histopathology of liver tissues. (a) Liver section of normal control rats showing normal architecture; (b) Liver section of CCl4 treated rats showing massive fatty changes, cellular necrosis, vacuolization, and ballooning degeneration; (c) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSC showing mild fatty vacuolation and cellular necrosis; (d) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSC showing cellular regeneration, mild degree of cell necrosis, and fatty vacuolation; (e) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSE showing few fatty vacuoles, restoration of hepatocytes, and lesser necrosis; (f) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSE showing minimal inflammatory cellular infiltration and almost near normal liver architecture; (g) Liver section of rats treated with CCl4 and quercetin showing recovery from the toxic effect with normal arrangement of hepatocytes with little evidence of fatty vacuoles and cellular necrosis (h) Liver histology of animals treated with silymarin showing normal histology with least parenchymal injury. (1-cellular necrosis; 2-vacuolization; 3-ballooning degeneration).

Mentions: The histological profile of liver sections of the control animals showed normal hepatic architecture with well-preserved cytoplasm, prominent nucleus, central vein, and compact arrangement of hepatocytes without fatty lobulation (Figure 5(a)). The liver sections of CCl4 treated animals showed hydropic changes in centrilobular hepatocytes with cell necrosis surrounded by neutrophils. Congestion of the central vein and sinusoids was seen with acute and chronic inflammatory cells infiltrating sinusoids mainly in the central zone. The midzonal and periportal hepatocytes showed vacuolization and fatty change (steatosis) which includes the intracellular accumulation of neutral fat (Figure 5(b)). The hepatocytes are distended with fat vacuoles due to increased deposition of intracellular lipids in liver section of DSC (100 mg kg−1 b.w.) administered animals (Figure 5(c)). In the liver sections of rats administered with DSC (300 mg kg−1 b.w.) showed mild fatty changes and mild sinusoidal congestion (Figure 5(d)). Animals administered with DSE (100 mg kg−1 b.w) exhibited significant liver protection against CCl4-induced liver damage. It is evident by the presence of hepatic cords with moderate fatty change and few inflammatory cells, and absence of necrosis (Figure 5(e)). The sections of liver taken from the animals treated with DSE (300 mg kg−1 b.w.) and quercetin (Figures 5(f) and 5(g) resp.) showed the normal hepatic architecture with presence of very few inflammatory cells and cell necrosis. Liver section of the rats treated with silymarin showed the presence of normal hepatic cords with few numbers of inflammatory cells and necrosis (Figure 5(h)).


Antioxidant and prophylactic effects of Delonix elata L., stem bark extracts, and flavonoid isolated quercetin against carbon tetrachloride-induced hepatotoxicity in rats.

Krishnappa P, Venkatarangaiah K - Biomed Res Int (2014)

Histopathology of liver tissues. (a) Liver section of normal control rats showing normal architecture; (b) Liver section of CCl4 treated rats showing massive fatty changes, cellular necrosis, vacuolization, and ballooning degeneration; (c) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSC showing mild fatty vacuolation and cellular necrosis; (d) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSC showing cellular regeneration, mild degree of cell necrosis, and fatty vacuolation; (e) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSE showing few fatty vacuoles, restoration of hepatocytes, and lesser necrosis; (f) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSE showing minimal inflammatory cellular infiltration and almost near normal liver architecture; (g) Liver section of rats treated with CCl4 and quercetin showing recovery from the toxic effect with normal arrangement of hepatocytes with little evidence of fatty vacuoles and cellular necrosis (h) Liver histology of animals treated with silymarin showing normal histology with least parenchymal injury. (1-cellular necrosis; 2-vacuolization; 3-ballooning degeneration).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4060769&req=5

fig5: Histopathology of liver tissues. (a) Liver section of normal control rats showing normal architecture; (b) Liver section of CCl4 treated rats showing massive fatty changes, cellular necrosis, vacuolization, and ballooning degeneration; (c) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSC showing mild fatty vacuolation and cellular necrosis; (d) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSC showing cellular regeneration, mild degree of cell necrosis, and fatty vacuolation; (e) Liver section of rats treated with CCl4 and 100 mg kg−1 of DSE showing few fatty vacuoles, restoration of hepatocytes, and lesser necrosis; (f) Liver section of rats treated with CCl4 and 300 mg kg−1 of DSE showing minimal inflammatory cellular infiltration and almost near normal liver architecture; (g) Liver section of rats treated with CCl4 and quercetin showing recovery from the toxic effect with normal arrangement of hepatocytes with little evidence of fatty vacuoles and cellular necrosis (h) Liver histology of animals treated with silymarin showing normal histology with least parenchymal injury. (1-cellular necrosis; 2-vacuolization; 3-ballooning degeneration).
Mentions: The histological profile of liver sections of the control animals showed normal hepatic architecture with well-preserved cytoplasm, prominent nucleus, central vein, and compact arrangement of hepatocytes without fatty lobulation (Figure 5(a)). The liver sections of CCl4 treated animals showed hydropic changes in centrilobular hepatocytes with cell necrosis surrounded by neutrophils. Congestion of the central vein and sinusoids was seen with acute and chronic inflammatory cells infiltrating sinusoids mainly in the central zone. The midzonal and periportal hepatocytes showed vacuolization and fatty change (steatosis) which includes the intracellular accumulation of neutral fat (Figure 5(b)). The hepatocytes are distended with fat vacuoles due to increased deposition of intracellular lipids in liver section of DSC (100 mg kg−1 b.w.) administered animals (Figure 5(c)). In the liver sections of rats administered with DSC (300 mg kg−1 b.w.) showed mild fatty changes and mild sinusoidal congestion (Figure 5(d)). Animals administered with DSE (100 mg kg−1 b.w) exhibited significant liver protection against CCl4-induced liver damage. It is evident by the presence of hepatic cords with moderate fatty change and few inflammatory cells, and absence of necrosis (Figure 5(e)). The sections of liver taken from the animals treated with DSE (300 mg kg−1 b.w.) and quercetin (Figures 5(f) and 5(g) resp.) showed the normal hepatic architecture with presence of very few inflammatory cells and cell necrosis. Liver section of the rats treated with silymarin showed the presence of normal hepatic cords with few numbers of inflammatory cells and necrosis (Figure 5(h)).

Bottom Line: Bioassay-guided fractionation of DSE has resulted in the isolation and characterization of quercetin.DSE and quercetin have shown significant prophylactic effects by restoring the liver function markers (AST, ALT, ALP, serum bilirubin, and total protein) and antioxidant enzymes (SOD, CAT, GPx, and GST).These results were proved to be hepatoprotective at par with silymarin and well supported by the histological observations of liver sections with distinct hepatic cells, and mild degree of fatty change and necrosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Post Graduate Studies and Research in Biotechnology, Kuvempu University, Shankaraghatta, Karnataka 577 451, India.

ABSTRACT
Delonix elata L. (Ceasalpinaceae), is widely used by the traditional medical practitioners of Karnataka, India, to cure jaundice, and bronchial and rheumatic problems. The objective of this study was to screen the in vitro antioxidant and hepatoprotective activity of the stem bark extracts against CCl4-induced liver damage in rats. Among different stem bark extracts tested, the ethanol extract (DSE) has shown significant in vitro antioxidant property in radicals scavenging, metal chelating, and lipid peroxidation inhibition assays. HPLC analysis of the DSE revealed the presence of known antioxidant molecules, namely, gallic acid, ellagic acid, coumaric acid, quercetin, and rutin. Bioassay-guided fractionation of DSE has resulted in the isolation and characterization of quercetin. DSE and quercetin have shown significant prophylactic effects by restoring the liver function markers (AST, ALT, ALP, serum bilirubin, and total protein) and antioxidant enzymes (SOD, CAT, GPx, and GST). These results were proved to be hepatoprotective at par with silymarin and well supported by the histological observations of liver sections with distinct hepatic cells, and mild degree of fatty change and necrosis. The results indicated that the DSE and quercetin were significant for prophylactic activity against CCl4-induced liver damage in rats. This activity could be attributed to the antioxidant constituents in the DSE and hence justified the ethnomedicinal claims.

Show MeSH
Related in: MedlinePlus