Free Radical Generation and Antioxidant Defense

Free Radical Generation and Antioxidant Defense

Free Radical Generation and Antioxidant Defense

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Free Radicals: Basics - Unstable Atoms Alert

  • Atoms or molecules with one or more unpaired electrons in their outer orbital; highly unstable and reactive.
  • Seek stability by oxidizing (stealing electrons from) or reducing (donating electrons to) other molecules.
  • Key Types:
    • Reactive Oxygen Species (ROS):
      • Superoxide ($O_2^{\cdot-}$)
      • Hydroxyl radical ($\cdot OH$) - extremely reactive.
      • Hydrogen peroxide ($H_2O_2$) (not a radical, but a key ROS)
    • Reactive Nitrogen Species (RNS):
      • Nitric oxide ($NO\cdot$)
      • Peroxynitrite ($ONOO^-$) Free Radical vs. Antioxidant: Electron Donation

⭐ The hydroxyl radical ($\cdot OH$) is the most reactive free radical, causing significant damage to DNA, lipids, and proteins. It has a very short half-life (nanoseconds).

FR Generation - Where Mischief Brews

Free Radical Sources and Cellular Damage

  • Free Radicals (FRs): Unstable molecules, unpaired electrons, cause cellular damage.
  • Endogenous Sources (Cellular Metabolism):
    • Mitochondrial ETC: Major site; 'leaks' superoxide ($O_2^{\cdot-}$).
    • NADPH Oxidase: Phagocytes for microbial killing (respiratory burst).
    • Xanthine Oxidase: Purine degradation to uric acid.
    • Fenton Reaction: $Fe^{2+} + H_2O_2 \rightarrow Fe^{3+} + \cdot OH + OH^-$. Iron/copper catalyzes hydroxyl radical ($\cdot OH$) formation.
    • Peroxisomal metabolism: e.g., fatty acid oxidation.
  • Exogenous Sources (Environmental):
    • Radiation: UV light, X-rays, gamma rays.
    • Pollutants: Air pollution, industrial chemicals.
    • Drugs: e.g., paracetamol overdose (OD).
    • Cigarette smoke: Rich in FRs.
    • Inflammation: Chronic inflammation ↑ FRs.

⭐ The hydroxyl radical, often formed via the Fenton reaction, is considered the most reactive and damaging oxygen-derived free radical.

FR Damage - Radicals on Rampage

Free radicals wreak havoc on cellular components:

  • Lipid Peroxidation: Polyunsaturated fatty acids (PUFAs) in membranes targeted.
    • Chain reaction: initiation, propagation, termination.
    • Markers: Malondialdehyde (MDA), 4-hydroxynonenal (4-HNE).
    • Damage: ↑ Membrane permeability, ↓ fluidity, cell lysis.
  • Protein Oxidation: Amino acid residues (Cys, Met) & protein backbone attacked.
    • Markers: Protein carbonyls, 3-nitrotyrosine.
    • Damage: Enzyme inactivation, protein aggregation & misfolding.
  • DNA Damage: Bases (esp. Guanine) & deoxyribose sugar affected.
    • Marker: $8-OHdG$ (8-hydroxy-2'-deoxyguanosine).
    • Damage: Mutations, DNA strand breaks, carcinogenesis. Free Radical Sources and Cellular Damage

⭐ 8-OHdG is a critical biomarker for oxidative DNA damage, frequently elevated in various cancers and degenerative diseases, making it a common exam topic related to free radical injury assessment and antioxidant efficacy studies.

Antioxidant Defenses - Body's Superheroes

  • Enzymatic Systems: Neutralize ROS. 📌 Mnemonic: Can Super Glue Protect? (Catalase, SOD, GPx)
    • Superoxide Dismutase (SOD): Converts $O_2^{\cdot-}$ to $H_2O_2$. (Cu-Zn, Mn, EC types).
    • Catalase (Peroxisomes): Degrades $H_2O_2$: $2H_2O_2 \rightarrow 2H_2O + O_2$.
    • Glutathione Peroxidase (GPx-Se): Selenium-dependent. Reduces $H_2O_2$/lipid peroxides: $H_2O_2 + 2GSH \rightarrow GSSG + 2H_2O$.
    • Glutathione Reductase: Regenerates GSH from GSSG (uses NADPH).
  • Non-Enzymatic Scavengers:
    • Glutathione (GSH): (\gamma)-Glu-Cys-Gly. Major intracellular antioxidant.
    • Vitamin E ((\alpha)-tocopherol): Lipid-soluble; protects membranes. Chain-breaking.
    • Vitamin C (Ascorbic acid): Water-soluble; regenerates Vit E; scavenges $O_2^{\cdot-}, OH^{\cdot}$.
    • Uric Acid: Plasma antioxidant (purine metabolism).
    • Bilirubin: Antioxidant (heme breakdown).
    • Others: Carotenoids, flavonoids, melatonin.

⭐ Glutathione (GSH) depletion ↑ oxidative stress susceptibility (e.g., Parkinson's, liver damage).

Oxidative Stress & Disease - Health Under Siege

  • Definition: Imbalance: ↑Reactive Oxygen/Nitrogen Species (ROS/RNS) vs. ↓Antioxidant capacity, causing cellular damage.
  • Key Damage Markers:
    • Lipids: Malondialdehyde (MDA).
    • Proteins: Carbonyls.
    • DNA: 8-hydroxy-2'-deoxyguanosine (8-OHdG).
  • Associated Diseases:
    • Atherosclerosis (LDL oxidation).
    • Neurodegeneration (Alzheimer's, Parkinson's).
    • Cancer (mutagenesis).
    • Ischemia-reperfusion injury.
    • Diabetes, chronic inflammation, aging.

⭐ 8-OHdG is a key biomarker for oxidative DNA damage, often elevated in various cancers and chronic diseases associated with oxidative stress.

High‑Yield Points - ⚡ Biggest Takeaways

  • Key Reactive Oxygen Species (ROS): superoxide (O₂⁻•), hydroxyl radical (•OH), hydrogen peroxide (H₂O₂).
  • Fenton reaction (Fe²⁺ + H₂O₂) generates highly reactive hydroxyl radicals (•OH).
  • Superoxide Dismutase (SOD) (Mn-mitochondrial, Cu/Zn-cytosolic) converts O₂⁻• to H₂O₂.
  • H₂O₂ is neutralized by Catalase and Selenium-dependent Glutathione Peroxidase.
  • Glutathione (GSH), a key antioxidant, is regenerated by Glutathione Reductase (uses NADPH).
  • Important antioxidant vitamins: Vitamin E (lipid-soluble), Vitamin C, and β-carotene.
  • Lipid peroxidation damages membranes; marker: Malondialdehyde (MDA).

Practice Questions: Free Radical Generation and Antioxidant Defense

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Where does the oxidation of drugs mainly take place?

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Flashcards: Free Radical Generation and Antioxidant Defense

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The enzyme responsible for conjugation of bilirubin, glucuronyl transferase is mainly located in the _____

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The enzyme responsible for conjugation of bilirubin, glucuronyl transferase is mainly located in the _____

endoplasmic reticulum (organelle)

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