ABSTRACT
Indigofera caerulea Roxb. is a well known shrub among native medical practitioners in folk medicine used for the treatment of jaundice, epilepsy, night blindness and snake bites. It is also reported to have antioxidant and antimicrobial properties. However its actual efficacy and hepatoprotective mechanism in particular is uncertain. Thus the present study investigates the hepatoprotective effect of the methanolic extract of I. caerulea Roxb. leaves (MIL) and elucidation of its mode of action against carbon tetrachloride (CCl4) induced liver injury in rats. HPLC analysis of MIL when carried out showed peaks close to standard ferulic acid and quercetin. Intragastric administration of MIL up to 2000 mg/kg bw, didn't show any toxicity and mortality in acute toxicity studies. During "in-vivo" study, hepatic injury was established by intraperitoneal administration of CCl4 3 ml/kg bw (30% CCl4 in olive oil; v/v) twice a week for 4 weeks in Sprague-Dawley rats. Further, hepatoprotective activity of MIL assessed using two different doses (100 and 200mg/kg bw) showed that intra-gastric administration of MIL (200mg/kg bw) significantly attenuates liver injury. Investigation of the underlying mechanism revealed that MIL treatment was capable of reducing inflammation by an antioxidant defense mechanism that blocks the activation of NF-κB as well as inhibits the release of proinflammatory cytokine TNF-α and IL-1ß. The results suggest that MIL has a significant hepatoprotective activity which might be due to the presence of phytochemicals namely analogues of ferulic acid and other phytochemicals which together may suppress the inflammatory signaling pathways and promote hepatoprotective activity against CCl4 intoxicated liver damage.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Carbon Tetrachloride/toxicity , Chemical and Drug Induced Liver Injury/prevention & control , Cytokines/immunology , Indigofera/chemistry , NF-kappa B/immunology , Plant Extracts/therapeutic use , Animals , Anti-Inflammatory Agents/isolation & purification , Chemical and Drug Induced Liver Injury/etiology , Chemical and Drug Induced Liver Injury/immunology , Chemical and Drug Induced Liver Injury/pathology , Cytokines/blood , Female , Immunohistochemistry , Liver/drug effects , Liver/immunology , Liver/pathology , Liver Function Tests , Male , NF-kappa B/blood , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Rats, Sprague-DawleyABSTRACT
Here, a large fraction of plant mitochondrial actin was found to be resistant to protease and high-salt treatments, suggesting it was protected by mitochondrial membranes. A portion of this actin became sensitive to protease or high-salt treatment after removal of the mitochondrial outer membrane, indicating that some actin is located inside the mitochondrial outer membrane. The import of an actin-green fluorescent protein (GFP) fusion protein into the mitochondria in a transgenic plant, actin:GFP, was visualized in living cells and demonstrated by flow cytometry and immunoblot analyses. Polymerized actin was found in mitochondria of actin:GFP plants and in mung bean (Vigna radiata). Notably, actin associated with mitochondria purified from early-developing cotyledons during seed germination was sensitive to high-salt and protease treatments. With cotyledon ageing, mitochondrial actin became more resistant to both treatments. The progressive import of actin into cotyledon mitochondria appeared to occur in concert with the conversion of quiescent mitochondria into active forms during seed germination. The binding of actin to mitochondrial DNA (mtDNA) was demonstrated by liquid chromatography-tandem mass spectrometry analysis. Porin and ADP/ATP carrier proteins were also found in mtDNA-protein complexes. Treatment with an actin depolymerization reagent reduced the mitochondrial membrane potential and triggered the release of cytochrome C. The potential function of mitochondrial actin and a possible actin import pathway are discussed.
Subject(s)
Actin Cytoskeleton/metabolism , Actins/metabolism , Mitochondria/metabolism , Phaseolus/metabolism , Amino Acid Sequence , Cotyledon/genetics , Cotyledon/metabolism , Cotyledon/ultrastructure , Cytochromes c/metabolism , DNA, Mitochondrial/genetics , Germination/drug effects , Mitochondrial Membranes/metabolism , Mitochondrial Proteins/metabolism , Molecular Sequence Data , Nucleoproteins/metabolism , Peptide Hydrolases/pharmacology , Phaseolus/drug effects , Phaseolus/genetics , Phaseolus/ultrastructure , Plants, Genetically Modified , Porins/metabolism , Potassium Chloride/pharmacology , Protein Transport , Recombinant Fusion Proteins , Seedlings/genetics , Seedlings/metabolism , Seedlings/ultrastructure , Seeds/drug effects , Seeds/genetics , Seeds/metabolism , Seeds/ultrastructureABSTRACT
UNLABELLED: The human Y chromosome is the sex determining chromosome. The number of proteins associated with this chromosome is 196 and 107 of the 196 proteins have yet not been characterised. Here, we describe the analysis of these 107 proteins by computing various physicochemical properties using sequence and predicted structural data to elucidate molecular function. We present the derived data in the form a form a database made freely available for download, review, refinement and update. AVAILABILITY: http://puratham.googlepages.com/ or http://puratham.googlepages.com/ftpconnection.
