Anat Cell Biol.  2011 Sep;44(3):218-225. 10.5115/acb.2011.44.3.218.

The high dosage of earthworm (Eisenia andrei) extract decreases cell proliferation and neuroblast differentiation in the mouse hippocampal dentate gyrus

• Yan BC
• Yoo KY
• Park JH
• Lee CH
• Choi JH
• Won MH

• KMID: 1447434
• DOI: http://doi.org/10.5115/acb.2011.44.3.218

Abstract

Earthworm extract has shown anticancer characteristics. In the present study, we examined the effect of chronic treatment with a high dose of earthworm (Eisenia andrei) extract (EE) on cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus (DG) of 3-week-old mice using 5-bromo-2'-deoxyuridine (BrdU) and Ki-67 immunohistochemistry for cell proliferation and doublecortin (DCX) immunohistochemistry for neuroblast differentiation, respectively. BrdU-, Ki-67-, and DCX-immunoreactive cells were easily detected in the subgranular zone of the DG in vehicle (saline)-treated mice. However, BrdU-, Ki-67-, and DCX-immunoreactive cells in the 500 mg/kg EE-treated mice decreased distinctively compared to those in the vehicle-treated mice. In addition, brain-derived neurotrophic factor (BDNF) immunoreactivity and its protein level decreased markedly in the DG of the EE-treated group compared to those in the vehicle-treated group. These results indicate that chronic treatment with high dose EE decreased cell proliferation and neuroblast differentiation, and that BDNF immunoreactivity decreased in the DG of EE-treated mice.

Keyword

Neurogenesis; Subgranular zone; Eisenia andrei extract; Neurotrophic factor

MeSH Terms

Animals
Brain-Derived Neurotrophic Factor
Bromodeoxyuridine
Cell Proliferation
Dentate Gyrus
Immunohistochemistry
Mice
Neurogenesis
Oligochaeta
Brain-Derived Neurotrophic Factor
Bromodeoxyuridine

Figure

• 

  Fig. 1 Immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) in the hippocampal dentate gyrus (DG) of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. BrdU+ cells (arrows) in the DG of the EE-treated group were lower than those in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of BrdU+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

• 

  Fig. 2 Immunohistochemistry for Ki-67 in the hippocampal dentate gyrus of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. Fewer Ki-67+ cells (arrows) were evident in the EE-treated group than those in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of Ki-67+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

• 

  Fig. 3 Immunohistochemistry for doublecortin (DCX) in the hippocampal dentate gyrus of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. DCX+ neuroblasts (arrows) were abundant, and DCX+ processes were well stained in the vehicle-treated group. However, DCX immunoreactivity in the EE-treated group decreased markedly. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of DCX+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

• 

  Fig. 4 Immunohistochemistry for brain derived neurotrophic factor (BDNF) in the hippocampal dentate gyrus (DG) of the vehicle- (A) and earthworm extract (EE)-treated (B) groups. BDNF immunoreactivity in the EE-treated group was weaker than that in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (C) Western blot analysis of BDNF in the DG of the vehicle- and EE-treated groups. Relative optical density (ROD) as a percentage of the immunoblot band is presented (n=5/group; *P<0.05, significantly different from the vehicle-treated group). Bars indicate mean±SD.

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