Briefly, SV-SMC from 5 different patients are seeded into 24-well cell culture plates at a density of 1×10⁴ cells per well in full growth medium. Cells are incubated overnight and then quiesced in serum free medium for 3 days before transfer to full growth medium (10% FCS) containing 5 different statins (simvastatin, atorvastatin, fluvastatin, lovastatin, and pravastatin)at a range of concentrations. All statins are tested on cells from each individual patient. Medium and drugs are replaced after 2 days, and viable cell numbers are determined in triplicate wells after 4 days using Trypan Blue and a hemocytometer. The increase in cell number is calculated by subtracting the starting cell number (day 0) from the final cell number (day 4). Data are then normalized to control values (no statin) to correct for differences in proliferation rates between cells from different patients.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

To investigate the effect of atorvastatin on lipopolysaccharide (LPS)-induced inflammatory hypernociception, mice are pretreated orally with either atorvastatin, at doses of 1, 3, 10, 30 and 90 mg/kg or vehicle (PBS) once a day for 3 consecutive days. At 2 h after the last dose of atorvastatin, mice receive an i.pl. injection of LPS (100 ng/paw) or saline (vehicle for LPS). The animals are also treated with atorvastatin (30 mg/kg) for 1 or 2 days before LPS challenge. The hypernociceptive responses are assessed 0.5, 1, 3, 5, 7 and 24 h after LPS or saline i.pl. injections.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Santodomingo-Garzón T, et al. Atorvastatin inhibits inflammatory hypernociception. Br J Pharmacol. 2006 Sep;149(1):14-22.

  [2]. Turner NA, et al. Comparison of the efficacies of five different statins on inhibition of human saphenous vein smooth muscle cell proliferation and invasion. J Cardiovasc Pharmacol. 2007 Oct;50(4):458-61.

  [3]. Lei A, et al. Atorvastatin promotes the expansion of myeloid-derived suppressor cells and attenuates murine colitis. Immunology. 2016 Aug 22.

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134523-03-8

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Room temperature in continental US; may vary elsewhere