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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

Morphological alteration of the microglia stimulated by LPS, IFN-γ, oAβ alone or combined with IFN-γ. Glial cells treated with vehicle (Veh), 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1–10 μM oAβ (Aβ1, Aβ5, Aβ10) for 24 h. (a) Cells were fixed and subjected to immuno-staining of microglia by anti-Iba1 antibody (red), astrocyte by anti-GFAP antibody (green) and nuclei staining using Hoechst 33258 (blue). Results are repeated for three times, and represent photographs are shown. (b) Form factor of microglial morphology 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. Significant differences between the cells treated with vehicle and the other treatments are indicated by *, p < 0.05, ***, p < 0.001.
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Figure 5: Morphological alteration of the microglia stimulated by LPS, IFN-γ, oAβ alone or combined with IFN-γ. Glial cells treated with vehicle (Veh), 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1–10 μM oAβ (Aβ1, Aβ5, Aβ10) for 24 h. (a) Cells were fixed and subjected to immuno-staining of microglia by anti-Iba1 antibody (red), astrocyte by anti-GFAP antibody (green) and nuclei staining using Hoechst 33258 (blue). Results are repeated for three times, and represent photographs are shown. (b) Form factor of microglial morphology 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. Significant differences between the cells treated with vehicle and the other treatments are indicated by *, p < 0.05, ***, p < 0.001.

Mentions: The morphological change of the ramified microglia in the primary mixed glial culture was further employed to examine the effect of Aβ on microglia activation. The microglia co-cultured with astrocyte displayed ramified processes with a small cell body and several long processes. The ramified morphology was altered to amoeboid as the cell was activated by 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1 to 10 μM oAβ (Figure 5a). To quantify the morphological alteration of microglia, the form factor of single cell was calculated as described in the Method section. The results showed that LPS and IFN-γ significantly increased the form factor of microglia from 0.29 ± 0.07 to 0.71 ± 0.04 and 0.62 ± 0.02, respectively (Figure 5b). oAβ, at 1 and 5 μM, increase the form factor of microglia from 0.29 ± 0.07 to 0.42 ± 0.10 and 0.77 ± 0.16, respectively. On the other hand, the morphology of astrocyte was altered by LPS, IFN-γ and oAβ from ramified to broaden processes or spreading lamella shape (Figure 5a).


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)

Morphological alteration of the microglia stimulated by LPS, IFN-γ, oAβ alone or combined with IFN-γ. Glial cells treated with vehicle (Veh), 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1–10 μM oAβ (Aβ1, Aβ5, Aβ10) for 24 h. (a) Cells were fixed and subjected to immuno-staining of microglia by anti-Iba1 antibody (red), astrocyte by anti-GFAP antibody (green) and nuclei staining using Hoechst 33258 (blue). Results are repeated for three times, and represent photographs are shown. (b) Form factor of microglial morphology 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. Significant differences between the cells treated with vehicle and the other treatments are indicated by *, p < 0.05, ***, p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Morphological alteration of the microglia stimulated by LPS, IFN-γ, oAβ alone or combined with IFN-γ. Glial cells treated with vehicle (Veh), 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1–10 μM oAβ (Aβ1, Aβ5, Aβ10) for 24 h. (a) Cells were fixed and subjected to immuno-staining of microglia by anti-Iba1 antibody (red), astrocyte by anti-GFAP antibody (green) and nuclei staining using Hoechst 33258 (blue). Results are repeated for three times, and represent photographs are shown. (b) Form factor of microglial morphology 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. Significant differences between the cells treated with vehicle and the other treatments are indicated by *, p < 0.05, ***, p < 0.001.
Mentions: The morphological change of the ramified microglia in the primary mixed glial culture was further employed to examine the effect of Aβ on microglia activation. The microglia co-cultured with astrocyte displayed ramified processes with a small cell body and several long processes. The ramified morphology was altered to amoeboid as the cell was activated by 10 ng/ml LPS, 0.5 ng/ml IFN-γ (IFN), or 1 to 10 μM oAβ (Figure 5a). To quantify the morphological alteration of microglia, the form factor of single cell was calculated as described in the Method section. The results showed that LPS and IFN-γ significantly increased the form factor of microglia from 0.29 ± 0.07 to 0.71 ± 0.04 and 0.62 ± 0.02, respectively (Figure 5b). oAβ, at 1 and 5 μM, increase the form factor of microglia from 0.29 ± 0.07 to 0.42 ± 0.10 and 0.77 ± 0.16, respectively. On the other hand, the morphology of astrocyte was altered by LPS, IFN-γ and oAβ from ramified to broaden processes or spreading lamella shape (Figure 5a).

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