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. 1997 Sep;41(3):344–353. doi: 10.1136/gut.41.3.344

Mitochondrial damage: a possible mechanism of the "topical" phase of NSAID induced injury to the rat intestine

S Somasundaram 1, S Rafi 1, J Hayllar 1, G Sigthorsson 1, M Jacob 1, A Price 1, A Macpherson 1, T Mahmod 1, D Scott 1, J Wrigglesworth 1, I Bjarnason 1
PMCID: PMC1891477  PMID: 9378390

Abstract

Background—The "topical" effect of non-steroidal anti-inflammatory drugs (NSAIDs) seems to be an important cause of NSAID induced gastrointestinal damage. 
Aim—To examine the possible mechanism of the "topical" phase of damage in the small intestine. 
Methods—Electron microscopy and subcellular organelle marker enzyme studies were done in rat small intestine after oral administration of indomethacin (doses varied between 5 and 30 mg/kg). The effect of conventional and non-acidic NSAIDs on rat liver mitochondrial respiration was measured in vitro in a Clarke-type oxygen electrode. 
Results—The subcellular organelle marker enzymes showed mitochondrial and brush border involvement within an hour of indomethacin administration. Electron microscopy showed dose dependent mitochondrial changes following indomethacin administration consistent with uncoupling of oxidative phosphorylation (or inhibition of electron transport) which were indistinguishable from those seen with the uncoupler dinitrophenol. Parenteral indomethacin caused similar changes, but not in rats with ligated bile ducts. A range of NSAIDs, but not paracetamol or non-acidic NSAIDs which have a favourable gastrointestinal tolerability profile, uncoupled oxidative phosphorylation in vitro at micromolar concentrations and inhibited respiration at higher concentrations. In vivo studies with nabumetone and aspirin further suggested that uncoupling or inhibition of electron transport underlies the "topical" phase of NSAID induced damage. 
Conclusion—Collectively, these studies suggest that NSAID induced changes in mitochondrial energy production may be an important component of the "topical" phase of damage induction. 



Keywords: intestinal inflammation; intestinal toxicity; mitochondrial function; drug induced toxicity

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Figure 1 .

Figure 1

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: Chemical structure of the NSAIDs and paracetamol used in the study. 


Figure 2 .

Figure 2

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: Representative electron transmission micrographs of rat small intestine (jejunum, 30 cm distal to the ligament of Treitz) demonstrating (A) normal enterocyte mitochondria, (B) vacuolisation of mitochondria one hour after indomethacin administration, (C) vacuolisation of mitochondria one hour after the administration of the uncoupler dinitrophenol, and (D) normal enterocyte mitochondria one hour after nabumetone administration. Note normal brush border appearances. Original magnification, ×10 000. 


Figure 3 .

Figure 3

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: Indomethacin, aspirin, naproxen, and piroxicam stimulate mitochondrial respiration (uncouple oxidative phosphorylation) at concentrations between 30 and 1500 µM, depending on the particular NSAID, and inhibit respiration at higher concentrations. Paracetamol, a non-NSAID analgesic control, reduces respiration, without an uncoupling effect. Each data point represents the mean (SE) of 3-5 experiments, performed on different days.


Figure 4 .

Figure 4

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: Mitochondrial preparation was as in fig 3, but with the addition of 0.1 µM FCCP, a potent uncoupler, prior to the addition of the drugs. (A) Glutamate/malate and (B) succinate (in the presence of rotenone) were used as electron donors for complexes I and II, respectively, at different concentrations of NSAIDs as shown in the figure. Indomethacin, aspirin, naproxen, and paracetamol all inhibit complexes I and II at different concentrations in a dose dependent manner. Each point represents the mean (SE) of 3-5 separate experiments, performed on different days. 


Figure 5 .

Figure 5

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: (A) The non-acidic pro-NSAID, nabumetone, had no significant effects on mitochondrial respiration whereas its active component, 6-MNA, stimulated respiration in a similar fashion to the other NSAIDs. (B) Similarly, flurbiprofen stimulated mitochondrial respiration whereas its chemical modification to a nitroxybutyl ester or a flurbiprofen dimer, renders the drugs incapable of uncoupling oxidative phosphorylation. 


Figure 6 .

Figure 6

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: Representative electron transmission micrographs of rat small intestine following intraperitoneal indomethacin (20 mg/kg) in (A) a rat with a ligated bile duct showing normal intestinal mitochondria with discrete cristae and (B) a sham operated animal showing disruption of mitochondrial cristae, vacuolisation and occasional disruption of both the outer and inner mitochondrial membranes. Endoplasmic reticulum is also dilated. Original magnification, × 30 000. 


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