Evaluation of therapeutic potential of cichorium intybus using high throughput computational and heavy metals intoxicated rat models

Abstract

This thesis research work was undertaken to assess the therapeutic potential of Cichorium intybus newlineusing high throughput in silico, in vitro phytochemical and antioxidant analysis and in vivo study for lead and newlinenickel induced subchronic toxicity and its amelioration in rats. In silico study revealed that selected newlinephytoconstituents of Cichorium intybus had good oral bioavailability and rutin, cichoric acid, chlorogenic newlineacid, lactopicrin and quercetin had highest binding affinity for CYP3A1 protein. Aqueous extract of newlineCichorium intybus leaves has highest percent yield followed by hydroethanolic extract (70%). Minerals like newlineCa+2, Mg+2 and Fe+2 were found in higher concentration than others in chicory leaf powder. Total flavonoid newlineand phenolic content were found in higher concentration in hydroethanolic than aqueous and methanolic newlineextracts of Cichorium intybus leaves. In vitro evaluation of antioxidant property of different extracts by DPPH, newlineABTS and nitric oxide free radical scavenging assay showed that hydroethanolic extract of Cichorium intybus newlineleaves (HECL) had minimum IC50 value and highest antioxidant property as compared to aqueous and newlinemethanolic extracts. For in vivo study, forty two male Wistar rats were divided equally and randomly into newlineseven groups. Group I served as control and other groups were given orally lead at 50 mg/kg b wt in group II, newlinenickel at 40 mg/kg b wt in group III, lead at 50 mg/kg b wt + nickel at 40 mg/kg b wt in group IV, lead at 50 newlinemg/kg b wt + HECL at 250 mg/kg b wt in group V, nickel at 40 mg/kg b wt + HECL at 250 mg/kg b wt in newlinegroup VI, lead at 50 mg/kg b wt + nickel at 40 mg/kg b wt + HECL at 250 mg/kg b wt in group VII, newlinerespectively, daily for 90 days. A significant (plt0.05) reduction was observed in body weight and feed intake newlinewhile mild to moderate changes in absolute organ weight in lead and nickel alone and in combination exposed newlinerats whereas, HECL treatments did not show any significant changes in body weight and feed intake. A newlinesignificant (plt0.05) reduction in TEC, TLC, Hb, PCV and platelet counts was observed in lead and nickel newlinealone and combined groups whereas a significant (plt0.05) improvement in TEC, TLC, Hb, PCV and platelet newlinecount was observed after treatment with HECL. A significant (plt0.05) increase in serum lipid profile and newlineserum ALT, AST, ALP, LDH and GGT activities while HDL was found to be decreased in lead and nickel newlinealone and combined groups, however, these values were ameliorated in HECL treated groups. Serum newlinecreatinine and BUN level were increased significantly (plt0.05) in lead and nickel exposed rats while newlinecombined group showed more increase in concentration of these parameters which were significantly (plt0.05) newlinerestored by the HECL treatment. A significant (plt0.05) increase in erythrocyte and tissue lipid peroxidation newlinewas observed while decrease in GSH content, SOD, CAT, GPx and GST activities in lead and nickel alone newlineand in combination groups whereas a significant (plt0.05) improvement was observed in these parameters newlinefollowing simultaneous medication with HECL for 90 days. Exposure to lead and nickel alone and in newlinecombination induced DNA fragmentation in liver whereas treatment with HECL showed partial improvement. newlinemRNA expression of CYP3A1 gene significantly (plt0.05) inhibited in lead and nickel alone and in newlinecombination groups whereas treatment with HECL significantly (plt0.05) restored mRNA expression of newlineCYP3A1 gene in lead and nickel alone groups. Testicular injury biomarkers (LDH and GGT) activities were newlinesignificantly (plt0.05) increased. A significant (plt0.05) decrease in live sperm percent, sperm motility and newlinesperm count while increase in dead sperm percentage, HOST positive sperm and total number of abnormal newlinesperms due to lead and nickel toxicity and combined group showed reproductive toxicity whereas HECL newlinesignificantly (plt0.05) restored these parameters toward normalcy in lead and nickel alone treatment groups. newlineTesticular injury biomarker (LDH and GGT) activity was significantly (plt0.05) increased in lead, nickel alone newlineand combined exposed groups whereas HECL significantly (plt0.05) restored these enzyme activities. newlineHistopathology changes in liver, kidney and testes and ultrastructure changes in liver by lead and newlinenickel were also reversed by treatment with HECL. newlineIt is concluded from this investigation that HECL at 250 mg/kg b wt reversed lead and nickel orally newlineadministered at 50 and 40 mg/kg b wt alone and in combination, induced haemotoxic, hepatotoxic, newlinenephrotoxic, alteration of drug metabolizing enzyme gene expression and reproductive toxic effect in a 90 newlinedays study in rats. newline( newline