Thesis In Physiology(BSC)

Published in: Physiology
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    Daniel O

    • Dubai
    • 5 Years of Experience
    • Qualification: BSc General Science and Physiology, PGCE Education
    • Teaches: Physics, Biology, SAT
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This PPT will summarise the tutor's work with endocannabinoids on the effects of inflammation during his BSc in Physiology.

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    Daniel O Hara Supervisor: Dr. Michelle NUI Galway oÉ Gaillimh
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    Innate and adaptive immunity LPS - CD14, TLR4 and MD-2 LPS re-negative bacte ria LPS LEP inflammatory cytokine release LPS _-—-.-* CLJ14 NF-"ÄL3 acer-rvar:jon Irrrrr•rurre cells like a Hage a a eell pathway of innate irnmune RAP mechani sm
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    Aims of Study Investigate the effect of LPS on PGE2 & PGF2a levels in brain and plasma. Effect of enhanced anandamide levels using AM404 on PGE2 and PGF2a concentration in brain and plasma samples Investigate if enhanced endocannabinoid tone modulates LPS-induced changes in PGE2
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    Materials and Methods AM404 — (20mg/kg) LPS 100gg/kg Figure l, tVtØ OOOOOOOOOOOO OOOOOOOOOOOO 100gl standard + oooooooooooo D OOOOOOOOOOOO sample to each well E OOOOOOOOOOOO F 0 00 G OOOOOOOOOOOO H OOOOOOOOOOOO A diagram Of a 96-well plate divided into units Of 16 samples, or one rtm, Plasma and brain samples Stored at -800C Samples carefully prepared
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    Protein Concentration Bradford Assay Extraction Citric acid Ethyl acetate PGE2 Metabolite Assay
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    PGE Metabolite std surve Protein standard curve 0.5 0.4 = 0.3 0.2 0.1 0.0 o 105 100 95 o 90 85 80 500 1000 1500 Results 2000 3000 150 100 o o 50 0.1 120 100 80 60 40 O 1 10 Concentration (pg/ml) PGE2 Standard Curve 1000 2000 PGE metabolite (pg/ml) PGF2u Standard Curve 1000 2000 3000 PGF2a Conc (pg/ml) 100 4000 PGE2 Conc (pg/ml)
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    PGE2 and PGF2a levels in the plasma following LPS PGE2 Plasma 12 10 Saline LPS
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    PGE2 and PGF2a levels from brain samples following LPS PGE2 Brain (assay) 12 10 Saline LPS PGF2a Brain (lysis) 12 10 Saline PGF2a Brain (Assay) LPS 12 10 Saline P < 0.05 LPS Data expressed as Mean + SEM.
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    PGE2 and PGF2a in plasma and brain fo owing LPS - extraction protocol PGF2a Plasma 12 Saline LPS 12 10 Saline 12 Saline LPS PGF2a Brain LPS
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    PGE2 and PGF2a levels from plasma and brain samples treated with either saline or AM404 PGE2 12 Cong 8 6 4 2 Saline Brain AM404 12 10 Conc8( g/ml) 6 4 2 Saline 12 Conc I 8 6 4 2 Saline PGF2a Plasma A M404 AM404
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    LPS induced increase in PGE2 levels-in-the-- plasma, not observed following AM404 treatment PGE2 Metabolite Brain 12 10 8 6 4 2 O Saline + Vehicle AM404 + Vehicle LPS + Vehicle LPS+ AM404
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    Summary PGE2 but not PGF2a increased in plasma and brain following LPS AM404 does not alter PGE2 or PGF2a in brain & plasma LPS increased PGE2 metabolite levels in plasma and pre-treatment with AM404 attenuated this
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    Conclusion AM404 — possible immunosuppressant effect as reported in previous studies. Enhancing EC tone attenuates induced increase in PGE2 metabolite in plasma following LPS. No effect of LPS or EC was observed on PGF2a Perhaps the role of investigating endocannabinoid modulators in more detail will

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