Anat Cell Biol.  2016 Mar;49(1):34-49. 10.5115/acb.2016.49.1.34.

Protective effect of rosemary on acrylamide motor neurotoxicity in spinal cord of rat offspring: postnatal follow-up study

• Al-Gholam
• Nooh HZ
• El-Mehi AE
• El-Barbary AEM
• Fokar AZ

• KMID: 2308930
• DOI: http://doi.org/10.5115/acb.2016.49.1.34

Abstract

The direct interactive effects of rosemary and acrylamide on the development of motor neurons in the spinal cord remains unknown. Our goal is to confirm the protective effects of rosemary against motor neuronal degeneration induced by acrylamide in the developing postnatal rat spinal cord using a postnatal rat model. We assigned the offspring of treated female rats into control, rosemary; acrylamide group; and recovery groups. This work depended on clinical, histopathological, morphometrically, immunohistochemical and genetic methods. In the acrylamide group, we observed oxidation, motor neuron degeneration, apoptosis, myelin degeneration, neurofilament reduction, reactive gliosis. Whoever, concomitant rosemary intake and withdrawal of acrylamide modulate these effects. These findings proof that dietary rosemary can directly protect motor neuron against acrylamide toxicity in the mammalian developing spinal cord.

Keyword

Acrylamide; Motor neuron; Rosemary; Development; Spinal cord

MeSH Terms

Acrylamide*
Animals
Apoptosis
Female
Follow-Up Studies*
Gliosis
Humans
Models, Animal
Motor Neurons
Myelin Sheath
Rats*
Spinal Cord*
Acrylamide

Figure

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  Fig. 1 Assessment of weight gain with age in different groups.

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  Fig. 2 Developmental landmarks in different groups.

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  Fig. 3 Assessment of gait score changes with age in different groups.

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  Fig. 4 Hematoxylin and eosin-stained (×400) lumbar anterior horn transverse sections in rats with advancement of age. (A–E) Control rat shows apparent increase in number and size of basophilic differentiated motor neurons (N) and decrease in undifferentiated one. In addition to, decrease in different form of neuroglia (G) and scattered small capillaries (C). (F–I) Acrylamide rat shows decrease in differentiated and increase in degenerated neurons (esinophilic [e], pyknotic [p]). In addition to, neuropil hemorrhage (asterisk) vacuolation neuropil (v), dilated congested capillaries, and neurogliosis (G) and neuronophagia (dashed circle). Protected (J–M) and recovery (N) rats, with age advancement, shows show similar picture to that of the control with few vacuolation, degenerated neurons and dilated congested capillaries.

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  Fig. 5 Sliver stained (×1,000) lumbar anterior horn transverse sections in in rats with advancement of age. (A–E) Control rat shows the motor neurons with central vesicular nucleus, small eccentric nucleoli acquires small dense granules (N), long branched dendrites (arrowheads) and the myelinated nodded axons (curved arrow). Oligodendroglia (O), astrocyte (A), and microglia (m) are also increased. Immature neuron with indistinct nucleus (U). (F–I) Acrylamide rat shows increase in neuron central chromatolysis (ch), swollen demyelination axons axon, and short dendrites. The degenerated oligodendroglia (o), astrocytes (A), and microglia (m) are also increased. Protected (I–M) and recovery (N) rats shows similar picture to that of the control group with few reported central chromatolysis (ch) especially in recovery rat.

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  Fig. 6 Toluidine blue (×1,000) stained lumbar anterior horn transverse sections in in rats with advancement of age. (A–E) Control rat shows dramatically increase in Nissl granules. (F–I) Acrylamide rat shows reduction in Nissl granules. Protected (J–M) and recovery (N) rats show upregulation in Nissl granules.

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  Fig. 7 Mean area (%) of toluidine blue in different groups.

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  Fig. 8 Neurofilament (NF) neurofilament (×1,000) immune-stained lumbar anterior horn transverse sections in rats with advancement of age. (A–E) Control rat shows dramatically increase in NF neurofilament. (F–I) Acrylamide rat shows reduction in NF neurofilament. Protected (J–M) and recovery (N) rats show upregulation in NF neurofilament.

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  Fig. 9 Mean neurofilament content area (%) in different groups.

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  Fig. 10 Myelin basic protein (MBP) neurofilament (×1,000) immune-stained lumbar anterior horn transverse sections in in rats with advancement of age. (A–E) Control rat shows dramatically increase in MBP neurofilament. (F–I) Acrylamide rat shows reduction in MBP neurofilament. Protected (J–M) and recovery (N) rats show upregulation in MBP neurofilament.

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  Fig. 11 Mean myelin basic protein content area (%) in different groups.

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  Fig. 12 Ethidium bromide stained comet assay (×40) in motor neuron in lumbar anterior horn in rats with advancement of age. (A–E) Control rat with normal neurons. (F–I) Acrylamide rat in increase of damaged neurons. Protected (J–M) and recovery (N) rats with downregulation of damaged neurons.

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