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Cudrania cochinchinensis attenuates amyloid β protein-mediated microglial activation and promotes glia-related clearance of amyloid β protein.

Wang CJ, Chen CC, Tsay HJ, Chiang FY, Wu MF, Shiao YJ - J. Biomed. Sci. (2013)

Bottom Line: LPS and IFN-γ, but not Aβs, activated BV-2 cells to produce nitric oxide through an increase in inducible nitric oxide synthase (iNOS) expression without significant effects on cell viability of microglia. fAβ, but not oAβ, enhanced the IFN-γ-stimulated nitric oxide production and iNOS expression.The ethanol/water extracts of Cudrania cochinchinensis (CC-EW) and the purified isolated components (i.e. CCA to CCF) effectively reduced the nitric oxide production and iNOS expression stimulated by IFN-γ combined with fAβ.CC-EW and CCB effectively prohibit the Aβ-mediated morphological change of microglia.Furthermore, CC-EW and CCB effectively decreased Aβ deposition and remained Aβ in the conditioned medium suggesting the effect of CC-EW and CCB on promoting Aβ clearance.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Geriatrics, Cheng Hsin General Hospital, Taipei, Taiwan.

ABSTRACT

Background: Microglial inflammation may significantly contribute to the pathology of Alzheimer's disease. To examine the potential of Cudrania cochinchinensis to ameliorate amyloid β protein (Aβ)-induced microglia activation, BV-2 microglial cell line, and the ramified microglia in the primary glial mixed cultured were employed.

Results: Lipopolysaccharide (LPS), Interferon-γ (IFN-γ), fibrillary Aβ (fAβ), or oligomeric Aβ (oAβ) were used to activate microglia. LPS and IFN-γ, but not Aβs, activated BV-2 cells to produce nitric oxide through an increase in inducible nitric oxide synthase (iNOS) expression without significant effects on cell viability of microglia. fAβ, but not oAβ, enhanced the IFN-γ-stimulated nitric oxide production and iNOS expression.The ethanol/water extracts of Cudrania cochinchinensis (CC-EW) and the purified isolated components (i.e. CCA to CCF) effectively reduced the nitric oxide production and iNOS expression stimulated by IFN-γ combined with fAβ. On the other hand, oAβ effectively activated the ramified microglia in mixed glial culture by observing the morphological alteration of the microglia from ramified to amoeboid. CC-EW and CCB effectively prohibit the Aβ-mediated morphological change of microglia. Furthermore, CC-EW and CCB effectively decreased Aβ deposition and remained Aβ in the conditioned medium suggesting the effect of CC-EW and CCB on promoting Aβ clearance. Results are expressed as mean ± S.D. and were analyzed by ANOVA with post-hoc multiple comparisons with a Bonferroni test.

Conclusions: The components of Cudrania cochinchinensis including CC-EW and CCB are potential for novel therapeutic intervention for Alzheimer's disease.

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Related in: MedlinePlus

CC-EW and CCB inhibit oAβ-induced morphological alteration of microglia and reduce the Aβ level. Glia were treated with vehicle (A), 20 μg/ml CC-EW (AC20), 100 μg/ml CC-EW (AC100), or 20 μM CCB (ACB20) for 2 h, and then were treated with 5 μM oAβ for 24 h. (a) Cells were fixed and subjected to immuno-staining by anti-Iba1 antibody (red), anti-GFAP antibody and anti-Aβ antibody (green). Nuclei were stained using Hoechst 33258 (blue). Results were repeated for three times, and represent photograph are shown. (b) Microglial form factor were calculated as describe in the Method section. Results are means ± S.D. from three independent experiments and total nine cells of each group were analyzed. (c) The number of Aβ-immunoreactive deposit per field (250 μm × 250 μm) of view was calculated using Metamorph software, and presented in percentage of the cells treated with Aβ alone. (d) The conditioned medium was collected and subjected to Aβ1-42 ELISA assay. The data is presented in percentage of the cells treated with Aβ alone. Results are means ± S.D. from three independent experiments. Significant differences between oAβ group and other treated group are indicated by **, p < 0.01; ***, p < 0.001.
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Figure 6: CC-EW and CCB inhibit oAβ-induced morphological alteration of microglia and reduce the Aβ level. Glia were treated with vehicle (A), 20 μg/ml CC-EW (AC20), 100 μg/ml CC-EW (AC100), or 20 μM CCB (ACB20) for 2 h, and then were treated with 5 μM oAβ for 24 h. (a) Cells were fixed and subjected to immuno-staining by anti-Iba1 antibody (red), anti-GFAP antibody and anti-Aβ antibody (green). Nuclei were stained using Hoechst 33258 (blue). Results were repeated for three times, and represent photograph are shown. (b) Microglial form factor were calculated as describe in the Method section. Results are means ± S.D. from three independent experiments and total nine cells of each group were analyzed. (c) The number of Aβ-immunoreactive deposit per field (250 μm × 250 μm) of view was calculated using Metamorph software, and presented in percentage of the cells treated with Aβ alone. (d) The conditioned medium was collected and subjected to Aβ1-42 ELISA assay. The data is presented in percentage of the cells treated with Aβ alone. Results are means ± S.D. from three independent experiments. Significant differences between oAβ group and other treated group are indicated by **, p < 0.01; ***, p < 0.001.

Mentions: CC-EW and CCB inhibited the morphological alteration induced by oAβ and the IC50 is 30.00 ± 3.65 μg/ml and 10.02 ± 2.11 μM, respectively (Figure 6a, 6b; Table 1). On the other hand, CC-EW and CCB did not significantly affect the morphology of astrocytes (Figure 6a).


Cudrania cochinchinensis attenuates amyloid β protein-mediated microglial activation and promotes glia-related clearance of amyloid β protein.

Wang CJ, Chen CC, Tsay HJ, Chiang FY, Wu MF, Shiao YJ - J. Biomed. Sci. (2013)

CC-EW and CCB inhibit oAβ-induced morphological alteration of microglia and reduce the Aβ level. Glia were treated with vehicle (A), 20 μg/ml CC-EW (AC20), 100 μg/ml CC-EW (AC100), or 20 μM CCB (ACB20) for 2 h, and then were treated with 5 μM oAβ for 24 h. (a) Cells were fixed and subjected to immuno-staining by anti-Iba1 antibody (red), anti-GFAP antibody and anti-Aβ antibody (green). Nuclei were stained using Hoechst 33258 (blue). Results were repeated for three times, and represent photograph are shown. (b) Microglial form factor were calculated as describe in the Method section. Results are means ± S.D. from three independent experiments and total nine cells of each group were analyzed. (c) The number of Aβ-immunoreactive deposit per field (250 μm × 250 μm) of view was calculated using Metamorph software, and presented in percentage of the cells treated with Aβ alone. (d) The conditioned medium was collected and subjected to Aβ1-42 ELISA assay. The data is presented in percentage of the cells treated with Aβ alone. Results are means ± S.D. from three independent experiments. Significant differences between oAβ group and other treated group are indicated by **, p < 0.01; ***, p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750318&req=5

Figure 6: CC-EW and CCB inhibit oAβ-induced morphological alteration of microglia and reduce the Aβ level. Glia were treated with vehicle (A), 20 μg/ml CC-EW (AC20), 100 μg/ml CC-EW (AC100), or 20 μM CCB (ACB20) for 2 h, and then were treated with 5 μM oAβ for 24 h. (a) Cells were fixed and subjected to immuno-staining by anti-Iba1 antibody (red), anti-GFAP antibody and anti-Aβ antibody (green). Nuclei were stained using Hoechst 33258 (blue). Results were repeated for three times, and represent photograph are shown. (b) Microglial form factor were calculated as describe in the Method section. Results are means ± S.D. from three independent experiments and total nine cells of each group were analyzed. (c) The number of Aβ-immunoreactive deposit per field (250 μm × 250 μm) of view was calculated using Metamorph software, and presented in percentage of the cells treated with Aβ alone. (d) The conditioned medium was collected and subjected to Aβ1-42 ELISA assay. The data is presented in percentage of the cells treated with Aβ alone. Results are means ± S.D. from three independent experiments. Significant differences between oAβ group and other treated group are indicated by **, p < 0.01; ***, p < 0.001.
Mentions: CC-EW and CCB inhibited the morphological alteration induced by oAβ and the IC50 is 30.00 ± 3.65 μg/ml and 10.02 ± 2.11 μM, respectively (Figure 6a, 6b; Table 1). On the other hand, CC-EW and CCB did not significantly affect the morphology of astrocytes (Figure 6a).

Bottom Line: LPS and IFN-γ, but not Aβs, activated BV-2 cells to produce nitric oxide through an increase in inducible nitric oxide synthase (iNOS) expression without significant effects on cell viability of microglia. fAβ, but not oAβ, enhanced the IFN-γ-stimulated nitric oxide production and iNOS expression.The ethanol/water extracts of Cudrania cochinchinensis (CC-EW) and the purified isolated components (i.e. CCA to CCF) effectively reduced the nitric oxide production and iNOS expression stimulated by IFN-γ combined with fAβ.CC-EW and CCB effectively prohibit the Aβ-mediated morphological change of microglia.Furthermore, CC-EW and CCB effectively decreased Aβ deposition and remained Aβ in the conditioned medium suggesting the effect of CC-EW and CCB on promoting Aβ clearance.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Geriatrics, Cheng Hsin General Hospital, Taipei, Taiwan.

ABSTRACT

Background: Microglial inflammation may significantly contribute to the pathology of Alzheimer's disease. To examine the potential of Cudrania cochinchinensis to ameliorate amyloid β protein (Aβ)-induced microglia activation, BV-2 microglial cell line, and the ramified microglia in the primary glial mixed cultured were employed.

Results: Lipopolysaccharide (LPS), Interferon-γ (IFN-γ), fibrillary Aβ (fAβ), or oligomeric Aβ (oAβ) were used to activate microglia. LPS and IFN-γ, but not Aβs, activated BV-2 cells to produce nitric oxide through an increase in inducible nitric oxide synthase (iNOS) expression without significant effects on cell viability of microglia. fAβ, but not oAβ, enhanced the IFN-γ-stimulated nitric oxide production and iNOS expression.The ethanol/water extracts of Cudrania cochinchinensis (CC-EW) and the purified isolated components (i.e. CCA to CCF) effectively reduced the nitric oxide production and iNOS expression stimulated by IFN-γ combined with fAβ. On the other hand, oAβ effectively activated the ramified microglia in mixed glial culture by observing the morphological alteration of the microglia from ramified to amoeboid. CC-EW and CCB effectively prohibit the Aβ-mediated morphological change of microglia. Furthermore, CC-EW and CCB effectively decreased Aβ deposition and remained Aβ in the conditioned medium suggesting the effect of CC-EW and CCB on promoting Aβ clearance. Results are expressed as mean ± S.D. and were analyzed by ANOVA with post-hoc multiple comparisons with a Bonferroni test.

Conclusions: The components of Cudrania cochinchinensis including CC-EW and CCB are potential for novel therapeutic intervention for Alzheimer's disease.

Show MeSH
Related in: MedlinePlus