Anat Cell Biol.  2013 Sep;46(3):183-190. 10.5115/acb.2013.46.3.183.

Neuroprotective effects of tanshinone I from Danshen extract in a mouse model of hypoxia-ischemia

• Lee JC
• Park JH
• Park OK
• Kim IH
• Yan BC
• Ahn JH
• Kwon SH
• Choi JH
• Kim JD
• Won MH

• KMID: 2263144
• DOI: http://doi.org/10.5115/acb.2013.46.3.183

Abstract

Hypoxia-ischemia leads to serious neuronal damage in some brain regions and is a strong risk factor for stroke. The aim of this study was to investigate the neuroprotective effect of tanshinone I (TsI) derived from Danshen (Radix Salvia miltiorrhiza root extract) against neuronal damage using a mouse model of cerebral hypoxia-ischemia. Brain infarction and neuronal damage were examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin histochemistry, and Fluoro-Jade B histofluorescence. Pre-treatment with TsI (10 mg/kg) was associated with a significant reduction in infarct volume 1 day after hypoxia-ischemia was induced. In addition, TsI protected against hypoxia-ischemia-induced neuronal death in the ipsilateral region. Our present findings suggest that TsI has strong potential for neuroprotection against hypoxic-ischemic damage. These results may be used in research into new anti-stroke medications.

Keyword

Hypoxia-ischemia; Neuronal death; Neuroprotection; Radix Salvia miltiorrhiza

MeSH Terms

Animals
Brain
Brain Infarction
Diterpenes, Abietane
Drugs, Chinese Herbal
Eosine Yellowish-(YS)
Fluoresceins
Hematoxylin
Hypoxia-Ischemia, Brain
Mice
Neurons
Neuroprotective Agents
Risk Factors
Salvia miltiorrhiza
Stroke
Tetrazolium Salts
Diterpenes, Abietane
Drugs, Chinese Herbal
Eosine Yellowish-(YS)
Fluoresceins
Hematoxylin
Neuroprotective Agents
Tetrazolium Salts

Figure

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  Fig. 1 Structure of tanshinone I.

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  Fig. 2 Neurological deficits observed in the sham, vehicle-treated, and tanshinone I (TsI)-treated hypoxia-ischemia groups 24 hours after induction of hypoxia-ischemia. The TsI-treated hypoxia-ischemia group showed a significant reduction in neurological deficits (n=7 per group; **P<0.05, significantly different from the vehicle-treated hypoxia-ischemia group). The bars indicate mean±SEM.

• 

  Fig. 3 TTC staining of mouse brain slices of the sham (A), vehicle-treated (B), and tanshinone I (TsI)-treated (C) hypoxia-ischemia groups 24 hours after induction of hypoxia-ischemia. The vehicle-treated hypoxia-ischemia group shows severe infarction (unstained region, *); the TsI-treated hypoxia-ischemia group shows a significant reduction in infarction (arrow). Scale bar in (C)=5 mm (A-C). (D) Infarct volume (mm3) of the sham, vehicle- and TsI-treated hypoxia-ischemia groups 24 hours after hypoxia-ischemia. (n=7 per group; **P<0.05, significantly different from the vehicle-treated hypoxia-ischemia group). The bars indicate mean±SEM.

• 

  Fig. 4 Hematoxylin and eosin (H&E) staining in the contralateral (A, C, E) and ipsilateral (B, D, F) regions of the cerebral cortex of the sham (A, B), vehicle-treated (C, D), and tanshinone I (TsI)-treated (E, F) hypoxia-ischemia groups 24 hours after induction of hypoxia-ischemia. Many cells (arrows) in the ipsilateral region show eosinophilic cytoplasm in the vehicle-treated hypoxia-ischemia group; H&E-positive neurons in the TsI-treated hypoxia-ischemia group are similar to those in the sham group. Scale bar in (F)=50 µm (A-F). (G) Relative numeric analysis of H&E-positive cells in the ipsilateral region in the sham, vehicle-treated, and TsI-treated hypoxia-ischemia groups (n=7 per group; *P<0.05, significantly different from the sham group; **P<0.05, significantly different from the vehicle-treated hypoxia-ischemia group). The bars indicate mean±SEM.

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  Fig. 5 Fluoro-Jade B (F-J B) histofluorescence staining in the contralateral (A, C, E) and ipsilateral (B, D, F) regions of the cerebral cortex of the sham (A, B), vehicle-treated (C, D), and tanshinone I (TsI)-treated (E, F) groups. Many F-J B-positive cells (arrows) in the ipsilateral region are observed in the vehicle-treated group; a few F-J B-positive cells are observed in the TsI-treated group. Scale bar in (F)=50 µm (A-F). (G) Relative numeric analysis of F-J B-positive cells in the ipsilateral region in the sham, vehicle-treated, and TsI-treated hypoxia-ischemia groups (n=7 per group; **P<0.05, significantly different from the vehicle-treated hypoxia-ischemia group). The bars indicate mean±SEM.

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