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The Components of Flemingia macrophylla Attenuate Amyloid β-Protein Accumulation by Regulating Amyloid β-Protein Metabolic Pathway.

Lin YL, Tsay HJ, Liao YF, Wu MF, Wang CN, Shiao YJ - Evid Based Complement Alternat Med (2012)

Bottom Line: Therefore, the effects of F. macrophylla on Aβ production and degradation were studied.The effect of F. macrophylla on Aβ metabolism was detected using the cultured mouse neuroblastoma cells N2a transfected with human Swedish mutant APP (swAPP-N2a cells).The effects on Aβ degradation were evaluated on a cell-free system.

View Article: PubMed Central - PubMed

Affiliation: Division of Medicinal Chemistry, National Research Institute of Chinese Medicine, Taipei 112, Taiwan.

ABSTRACT
Flemingia macrophylla (Leguminosae) is a popular traditional remedy used in Taiwan as anti-inflammatory, promoting blood circulation and antidiabetes agent. Recent study also suggested its neuroprotective activity against Alzheimer's disease. Therefore, the effects of F. macrophylla on Aβ production and degradation were studied. The effect of F. macrophylla on Aβ metabolism was detected using the cultured mouse neuroblastoma cells N2a transfected with human Swedish mutant APP (swAPP-N2a cells). The effects on Aβ degradation were evaluated on a cell-free system. An ELISA assay was applied to detect the level of Aβ1-40 and Aβ1-42. Western blots assay was employed to measure the levels of soluble amyloid precursor protein and insulin degrading enzyme (IDE). Three fractions of F. macrophylla modified Aβ accumulation by both inhibiting β-secretase and activating IDE. Three flavonoids modified Aβ accumulation by activating IDE. The activated IDE pool by the flavonoids was distinctly regulated by bacitracin (an IDE inhibitor). Furthermore, flavonoid 94-18-13 also modulates Aβ accumulation by enhancing IDE expression. In conclusion, the components of F. macrophylla possess the potential for developing new therapeutic drugs for Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus

The effect of the fractions of F. macrophylla on the level of secreted APP in swAPP770-transfected N2a cell. swAPP770-transfected N2a cells were treated with the fractions for 20 h at NTC of 10, 1, and 10 μg/mL, respectively. The level of 6E10 antibody-stained sAPP (i.e., sAPPα) and KPI antibody-stained sAPPs (i.e., sAPPα plus sAPPβ) in conditioned medium was determined by immunoblotting. The upper part is the representative image of immunoblot. The lower part is the relative level of 6E10 antibody-stained sAPPα (opened column) and KPI antibody-stained sAPPs (closed column). Results are means ± SD from three independent experiments. Significant differences between control and FM fractions-treated cells are indicated by *P < 0.05.
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fig7: The effect of the fractions of F. macrophylla on the level of secreted APP in swAPP770-transfected N2a cell. swAPP770-transfected N2a cells were treated with the fractions for 20 h at NTC of 10, 1, and 10 μg/mL, respectively. The level of 6E10 antibody-stained sAPP (i.e., sAPPα) and KPI antibody-stained sAPPs (i.e., sAPPα plus sAPPβ) in conditioned medium was determined by immunoblotting. The upper part is the representative image of immunoblot. The lower part is the relative level of 6E10 antibody-stained sAPPα (opened column) and KPI antibody-stained sAPPs (closed column). Results are means ± SD from three independent experiments. Significant differences between control and FM fractions-treated cells are indicated by *P < 0.05.

Mentions: The anabolic pathway of Aβ may also be affected by the fractions of F. macrophylla which resulted in the decrease of extracellular Aβ. Two categories of soluble APP (sAPP) including α-secretase-derived sAPP (sAPPα) and β-secretase-derived sAPP (sAPPβ) may be detected in the swAPP-N2a-conditioned medium. Two antibodies were used to detect these two sAPPs. The anti-Aβ1-17 (6E10) antibody may recognize sAPPα (this fragment contains Aβ1-17), and the anti-APP (KPI domain) antibody may recognize both sAPPα and sAPPβ on immunoblot. The result showed that 6E10 antibody-recognized sAPPα was not significantly affected by the fractions of F. macrophylla. By contrast, the anti-APP (KPI domain) antibody-recognized sAPPα and sAPPβ were significantly decreased. The fractions of EtOH, EA-47, and B50M decreased the level of sAPPs to 76.73 ± 9.11%, 78.99 ± 7.02%, and 79.44 ± 8.48%, respectively. The result indicated that the fractions may inhibit the activity of β-secretase and then decrease the level of sAPPβ (Figure 7), which may reflect the effects of these fractions on attenuating Aβ accumulation.


The Components of Flemingia macrophylla Attenuate Amyloid β-Protein Accumulation by Regulating Amyloid β-Protein Metabolic Pathway.

Lin YL, Tsay HJ, Liao YF, Wu MF, Wang CN, Shiao YJ - Evid Based Complement Alternat Med (2012)

The effect of the fractions of F. macrophylla on the level of secreted APP in swAPP770-transfected N2a cell. swAPP770-transfected N2a cells were treated with the fractions for 20 h at NTC of 10, 1, and 10 μg/mL, respectively. The level of 6E10 antibody-stained sAPP (i.e., sAPPα) and KPI antibody-stained sAPPs (i.e., sAPPα plus sAPPβ) in conditioned medium was determined by immunoblotting. The upper part is the representative image of immunoblot. The lower part is the relative level of 6E10 antibody-stained sAPPα (opened column) and KPI antibody-stained sAPPs (closed column). Results are means ± SD from three independent experiments. Significant differences between control and FM fractions-treated cells are indicated by *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3376484&req=5

fig7: The effect of the fractions of F. macrophylla on the level of secreted APP in swAPP770-transfected N2a cell. swAPP770-transfected N2a cells were treated with the fractions for 20 h at NTC of 10, 1, and 10 μg/mL, respectively. The level of 6E10 antibody-stained sAPP (i.e., sAPPα) and KPI antibody-stained sAPPs (i.e., sAPPα plus sAPPβ) in conditioned medium was determined by immunoblotting. The upper part is the representative image of immunoblot. The lower part is the relative level of 6E10 antibody-stained sAPPα (opened column) and KPI antibody-stained sAPPs (closed column). Results are means ± SD from three independent experiments. Significant differences between control and FM fractions-treated cells are indicated by *P < 0.05.
Mentions: The anabolic pathway of Aβ may also be affected by the fractions of F. macrophylla which resulted in the decrease of extracellular Aβ. Two categories of soluble APP (sAPP) including α-secretase-derived sAPP (sAPPα) and β-secretase-derived sAPP (sAPPβ) may be detected in the swAPP-N2a-conditioned medium. Two antibodies were used to detect these two sAPPs. The anti-Aβ1-17 (6E10) antibody may recognize sAPPα (this fragment contains Aβ1-17), and the anti-APP (KPI domain) antibody may recognize both sAPPα and sAPPβ on immunoblot. The result showed that 6E10 antibody-recognized sAPPα was not significantly affected by the fractions of F. macrophylla. By contrast, the anti-APP (KPI domain) antibody-recognized sAPPα and sAPPβ were significantly decreased. The fractions of EtOH, EA-47, and B50M decreased the level of sAPPs to 76.73 ± 9.11%, 78.99 ± 7.02%, and 79.44 ± 8.48%, respectively. The result indicated that the fractions may inhibit the activity of β-secretase and then decrease the level of sAPPβ (Figure 7), which may reflect the effects of these fractions on attenuating Aβ accumulation.

Bottom Line: Therefore, the effects of F. macrophylla on Aβ production and degradation were studied.The effect of F. macrophylla on Aβ metabolism was detected using the cultured mouse neuroblastoma cells N2a transfected with human Swedish mutant APP (swAPP-N2a cells).The effects on Aβ degradation were evaluated on a cell-free system.

View Article: PubMed Central - PubMed

Affiliation: Division of Medicinal Chemistry, National Research Institute of Chinese Medicine, Taipei 112, Taiwan.

ABSTRACT
Flemingia macrophylla (Leguminosae) is a popular traditional remedy used in Taiwan as anti-inflammatory, promoting blood circulation and antidiabetes agent. Recent study also suggested its neuroprotective activity against Alzheimer's disease. Therefore, the effects of F. macrophylla on Aβ production and degradation were studied. The effect of F. macrophylla on Aβ metabolism was detected using the cultured mouse neuroblastoma cells N2a transfected with human Swedish mutant APP (swAPP-N2a cells). The effects on Aβ degradation were evaluated on a cell-free system. An ELISA assay was applied to detect the level of Aβ1-40 and Aβ1-42. Western blots assay was employed to measure the levels of soluble amyloid precursor protein and insulin degrading enzyme (IDE). Three fractions of F. macrophylla modified Aβ accumulation by both inhibiting β-secretase and activating IDE. Three flavonoids modified Aβ accumulation by activating IDE. The activated IDE pool by the flavonoids was distinctly regulated by bacitracin (an IDE inhibitor). Furthermore, flavonoid 94-18-13 also modulates Aβ accumulation by enhancing IDE expression. In conclusion, the components of F. macrophylla possess the potential for developing new therapeutic drugs for Alzheimer's disease.

No MeSH data available.


Related in: MedlinePlus