Fig 1.
Molecular structures of TMPyP4 and TMPyP2.
Fig 2.
In vitro vs. in vivo modulation of Th expression by TMPyP4.
A, transcription assays with SH-SY5Y cells transfected with a reporter under the control of the rat 4.5kb Th promoter that are also treated with either saline, TMPyP2 or TMPyP4. Mean relative promoter activities are shown with error bars representing the standard deviation. ** indicates p<0.001, * indicates p = 0.02 and n.s. indicates not significant. B, Th transcript levels, as measured by quantitative RT-PCR, in the adrenal glands, olfactory bulb and midbrain of adult mice administered either saline, TMPyP2 or TMPyP4 (n = 8 for both male and female). Mean relative transcript levels are shown with error bars representing the standard error of the mean. ** indicates p<0.01, * indicates p = 0.03 and n.s. indicates not significant.
Table 1.
Physiological symptoms observed following a single injection with a high dose (40mg/kg) of either TMPyP2 or TMPyP4.
Fig 3.
TMPyP4 inhibits AChE activity.
A, oxidation of heme by Hemeoxygenase (HO) enzymes produces biliverdin and carbon monoxide. B, oxidation of TMPyP4 by HO enzymes is expected to generate either 4-formyl-1-methylpyridinium (4F-MP) or 4-carboxy-1-methylpyridinium (4C-MP). Both 4F-MP and 4C-MP have a structural resemblance to acetylcholine. C, fluorometric assay of AChE activity in presence of TMPyP2, TMPyP4, 4F-MP, 4C-MP and an established AChE inhibitor (Donepezil). D, pre-incubation of TMPyP4 with recombinant HO-2 prior rescued AChE activity in a concentration-dependent manner.
Fig 4.
Age and gender analysis of HO-1/2 expression levels in liver and skeletal muscle.
A-B, western blots for HO-1/2 and GAPDH in the liver and skeletal muscle, respectively, of young and old adult mice for both sexes. C and E, D and F, average relative western blot band intensities for HO-1/2 in liver and skeletal muscle tissue, respectively. Averages are from 3 independent trials for each age and gender in both liver and muscle tissue.
Fig 5.
Gender-based analysis of AChE activity.
A and B, fluorometric AChE activity assays with liver and skeletal muscle tissue lysates, respectively, from young adult mice (aged 1.5–3 months). For both males and females, data points are the combination of four individuals. AChE activities of tissue lysates are reported relative to the activity of an AChE standard supplied with the assay kit.