title: The effects of two common edible herbs, Ipomoea aquatica and Enhydra fluctuans, on cadmium-induced pathophysiology: a focus on oxidative defence and anti-apoptotic mechanism.
abstract: BACKGROUND
  Ipomoea aquatica (Convolvulaceae) and Enhydra fluctuans (Asteraceae), two aquatic vegetables, are traditionally used against heavy metal toxicity in traditional medicines in India. The present study aimed to explore the protective role of edible (aqueous) extracts of I. aquatica (AEIA) and E. fluctuans (AEEF) against Cd-intoxication.
  
  
  METHODS
  The extracts were chemically standardized by spectroscopic and HPLC analysis. The cytoprotective roles of AEIA and AEEF were measured on mouse hepatocytes. The effect on redox status were measured after incubating the hepatocytes with CdCl2 (30 μM) along with AEIA or AEEF (400 μg/ml). The effects on the expressions of apoptotic signal proteins were estimated. The protective roles of AEIA or AEEF were measured by in vivo assay in mice. Haematological, serum biochemical, tissue redox status, Cd bioaccumulation and histological parameters were evaluated to estimate the protective role of AEIA or AEEF (100 mg/kg) against CdCl2 (4 mg/kg) intoxication.
  
  
  RESULTS
  Phytochemical analysis revealed presence of substantial quantities of phenolics, flavonoids, saponins, carbohydrates and ascorbic acid in AEIA or AEEF. CdCl2 treated murine hepatocytes showed a gradual reduction of cell viability in a concentration dependent manner with an IC50 of ~30 μM. CdCl2 treated hepatocytes exhibited significantly enhanced levels (p < 0.01) of ROS production, lipid peroxidation, protein carbonylation and NADPH oxidase with concomitant depletion (p < 0.01) of antioxidant enzymes and GSH. However, AEIA or AEEF treatment along with CdCl2 significantly restored the aforementioned parameters in murine hepatocytes near to normalcy. Besides, AEIA or AEEF significantly counteracted (p < 0.05-0.01) with ROS mediated alteration of transcription levels of signal proteins viz. Bcl-2, BAD, Cyt-C, Caspases, Fas and Bid. In in vivo bioassay, CdCl2 treatment caused significantly high Cd bioaccumulation and oxidative stress in the liver, kidney, heart, brain and testes in mice. In addition, the haematological and serum biochemical parameters were significantly altered in the CdCl2 treated animals. Simultaneous administration of AEIA or AEEF could significantly restore the tested parameters to the near-normal status.
  
  
  CONCLUSIONS
  The extracts would offer the overall protective effect via counteracting with Cd mediated oxidative stress and/or promoting the elimination of Cd by chelating.
authors.affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. s.dewanjee@yahoo.com.
authors.forename: Tarun K
authors.initials: TK
authors.lastname: Dua
authors.affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. s.dewanjee@yahoo.com.
authors.forename: Saikat
authors.initials: S
authors.lastname: Dewanjee
authors.affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. s.dewanjee@yahoo.com.
authors.forename: Ritu
authors.initials: R
authors.lastname: Khanra
authors.affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. s.dewanjee@yahoo.com.
authors.forename: Niloy
authors.initials: N
authors.lastname: Bhattacharya
authors.affiliation: Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. s.dewanjee@yahoo.com.
authors.forename: Bhuvan
authors.initials: B
authors.lastname: Bhaskar
authors.affiliation: The Patent Office, Karachi, 74470, Pakistan. s.dewanjee@yahoo.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Zia-Ul-Haq
authors.affiliation: Department of Pharmacy, University of Salerno, 84084, Fisciano, Salerno, Italy. defeo@unisa.it.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Feo
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D019659:Asteraceae / Q000737:chemistry*; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D019256:Cadmium Chloride / Q000633:toxicity*; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D027741:Ipomoea / Q000737:chemistry*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D051379:Mice; D019255:NADPH Oxidases / Q000378:metabolism; D009928:Organ Specificity / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D064209:Phytochemicals / Q000494:pharmacology; D010936:Plant Extracts / Q000494:pharmacology*; D010945:Plants, Edible / Q000737:chemistry*; D050050:Protein Carbonylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D051025:bcl-Associated Death Protein / Q000378:metabolism
chemical_list: D000975:Antioxidants; D015415:Biomarkers; D064209:Phytochemicals; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D051025:bcl-Associated Death Protein; D019255:NADPH Oxidases; D048031:JNK Mitogen-Activated Protein Kinases; D005978:Glutathione; D019256:Cadmium Chloride