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Therapeutic Effect of Berberine on Huntington's Disease Transgenic Mouse Model.

Jiang W, Wei W, Gaertig MA, Li S, Li XJ - PLoS ONE (2015)

Bottom Line: We found that BBR can reduce the accumulation of mutant huntingtin in cultured cells.We found that BBR could promote the degradation of mutant huntingtin by enhancing autophagic function.Since BBR is an orally-taken drug that has been safely used to treat a number of diseases, our findings suggest that BBR can be tested on different HD animal models and HD patients to further evaluate its therapeutic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Genetics, Emory University School of Medicine, 615 Michael Street, Atlanta, GA 30322, United States of America; Graduate Program of Microbiology and Molecular Genetics, Emory University, Atlanta, GA 30322, United States of America.

ABSTRACT
Huntington disease (HD) represents a family of neurodegenerative diseases that are caused by misfolded proteins. The misfolded proteins accumulate in the affected brain regions in an age-dependent manner to cause late-onset neurodegeneration. Transgenic mouse models expressing the HD protein, huntingtin, have been widely used to identify therapeutics that may retard disease progression. Here we report that Berberine (BBR), an organic small molecule isolated from plants, has protective effects on transgenic HD (N171-82Q) mice. We found that BBR can reduce the accumulation of mutant huntingtin in cultured cells. More importantly, when given orally, BBR could effectively alleviate motor dysfunction and prolong the survival of transgenic N171-82Q HD mice. We found that BBR could promote the degradation of mutant huntingtin by enhancing autophagic function. Since BBR is an orally-taken drug that has been safely used to treat a number of diseases, our findings suggest that BBR can be tested on different HD animal models and HD patients to further evaluate its therapeutic effects.

No MeSH data available.


Related in: MedlinePlus

BBR reduced Htt aggregation in vitro in a dose and time-dependent manner.(A) Immunocytostaining images (10 X) of Htt-120Q- or Htt-20Q-transfected HEK293 cells that were treated with different concentrations of BBR (0, 5, 25, 50, 100 μM). (B) Western blot analysis of Htt-transfected cells treated with or without BBR at different concentrations. Aggregated Htt in the stacking gel was detected by mEM48 antibody. Soluble mutant Htt and its potential degraded products are also shown. (C) Densitometry analysis of the ratios of aggregated Htt or Htt-20Q to β-actin on western blots in (B). (D) Fluorescent microscopic images of Htt-120Q-transfected HEK293 cells that were treated with 50 μM BBR at 0 h, 12 h or 24 h post-transfection. (E) Cell-counting analysis of images obtained in (D) showing the percentage of aggregates relative to the total cells revealed by DAPI nuclear staining. (F) Western blotting of Htt-120Q transfected HEK293 cells treated with 50 μm BBR for different times showing aggregated Htt in the stacking gel. (G) Densitometry analysis of the ratios of mutant Htt to β-actin on western blots in (F). The quantitative data are presented as mean±SE.
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pone.0134142.g001: BBR reduced Htt aggregation in vitro in a dose and time-dependent manner.(A) Immunocytostaining images (10 X) of Htt-120Q- or Htt-20Q-transfected HEK293 cells that were treated with different concentrations of BBR (0, 5, 25, 50, 100 μM). (B) Western blot analysis of Htt-transfected cells treated with or without BBR at different concentrations. Aggregated Htt in the stacking gel was detected by mEM48 antibody. Soluble mutant Htt and its potential degraded products are also shown. (C) Densitometry analysis of the ratios of aggregated Htt or Htt-20Q to β-actin on western blots in (B). (D) Fluorescent microscopic images of Htt-120Q-transfected HEK293 cells that were treated with 50 μM BBR at 0 h, 12 h or 24 h post-transfection. (E) Cell-counting analysis of images obtained in (D) showing the percentage of aggregates relative to the total cells revealed by DAPI nuclear staining. (F) Western blotting of Htt-120Q transfected HEK293 cells treated with 50 μm BBR for different times showing aggregated Htt in the stacking gel. (G) Densitometry analysis of the ratios of mutant Htt to β-actin on western blots in (F). The quantitative data are presented as mean±SE.

Mentions: First, we wanted to see whether BBR would produce any effect on the accumulation of mutant Htt in transfected cells. BBR was added to the culture medium at concentrations of 0, 5, 25, 50, and 100 μM immediately after transfection of HEK293 cells with GFP-exon1 Htt containing 120Q (Htt-120Q). After 48 h of incubation with BBR, the cells were examined via fluorescent microscopy. The results showed a dose-dependent decrease of Htt-120Q aggregates, which were presented as puncta, with notable reduction at 50 μM BBR (Fig 1A). However, HEK293 cells transfected with the control GFP-exon1 Htt containing 20Q (Htt-20Q) did not exhibit any significant GFP signal reduction with even the highest concentration of BBR (100 μM), suggesting that BBR selectively reduces the accumulation of mutant Htt (Fig 1B) and since both Htt-20Q and Htt-120Q were both under a cytomegalovirus promoter, it also suggests that BBR does not impede transfection or promoter activity. The effect of BBR on reducing Htt aggregation was also shown by Western blotting that revealed aggregated Htt in the stacking gel. Quantification of the ratios of aggregated Htt to actin via densitometry also verified the reduction of Htt aggregates by BBR (Fig 1B and 1C) (for treatment with 50uM BBR, P = 0.028, T = 5.85, DF = 2). These results suggest that BBR can suppress the aggregate formation or the accumulation of mutant Htt (Fig 1A–1C).


Therapeutic Effect of Berberine on Huntington's Disease Transgenic Mouse Model.

Jiang W, Wei W, Gaertig MA, Li S, Li XJ - PLoS ONE (2015)

BBR reduced Htt aggregation in vitro in a dose and time-dependent manner.(A) Immunocytostaining images (10 X) of Htt-120Q- or Htt-20Q-transfected HEK293 cells that were treated with different concentrations of BBR (0, 5, 25, 50, 100 μM). (B) Western blot analysis of Htt-transfected cells treated with or without BBR at different concentrations. Aggregated Htt in the stacking gel was detected by mEM48 antibody. Soluble mutant Htt and its potential degraded products are also shown. (C) Densitometry analysis of the ratios of aggregated Htt or Htt-20Q to β-actin on western blots in (B). (D) Fluorescent microscopic images of Htt-120Q-transfected HEK293 cells that were treated with 50 μM BBR at 0 h, 12 h or 24 h post-transfection. (E) Cell-counting analysis of images obtained in (D) showing the percentage of aggregates relative to the total cells revealed by DAPI nuclear staining. (F) Western blotting of Htt-120Q transfected HEK293 cells treated with 50 μm BBR for different times showing aggregated Htt in the stacking gel. (G) Densitometry analysis of the ratios of mutant Htt to β-actin on western blots in (F). The quantitative data are presented as mean±SE.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520448&req=5

pone.0134142.g001: BBR reduced Htt aggregation in vitro in a dose and time-dependent manner.(A) Immunocytostaining images (10 X) of Htt-120Q- or Htt-20Q-transfected HEK293 cells that were treated with different concentrations of BBR (0, 5, 25, 50, 100 μM). (B) Western blot analysis of Htt-transfected cells treated with or without BBR at different concentrations. Aggregated Htt in the stacking gel was detected by mEM48 antibody. Soluble mutant Htt and its potential degraded products are also shown. (C) Densitometry analysis of the ratios of aggregated Htt or Htt-20Q to β-actin on western blots in (B). (D) Fluorescent microscopic images of Htt-120Q-transfected HEK293 cells that were treated with 50 μM BBR at 0 h, 12 h or 24 h post-transfection. (E) Cell-counting analysis of images obtained in (D) showing the percentage of aggregates relative to the total cells revealed by DAPI nuclear staining. (F) Western blotting of Htt-120Q transfected HEK293 cells treated with 50 μm BBR for different times showing aggregated Htt in the stacking gel. (G) Densitometry analysis of the ratios of mutant Htt to β-actin on western blots in (F). The quantitative data are presented as mean±SE.
Mentions: First, we wanted to see whether BBR would produce any effect on the accumulation of mutant Htt in transfected cells. BBR was added to the culture medium at concentrations of 0, 5, 25, 50, and 100 μM immediately after transfection of HEK293 cells with GFP-exon1 Htt containing 120Q (Htt-120Q). After 48 h of incubation with BBR, the cells were examined via fluorescent microscopy. The results showed a dose-dependent decrease of Htt-120Q aggregates, which were presented as puncta, with notable reduction at 50 μM BBR (Fig 1A). However, HEK293 cells transfected with the control GFP-exon1 Htt containing 20Q (Htt-20Q) did not exhibit any significant GFP signal reduction with even the highest concentration of BBR (100 μM), suggesting that BBR selectively reduces the accumulation of mutant Htt (Fig 1B) and since both Htt-20Q and Htt-120Q were both under a cytomegalovirus promoter, it also suggests that BBR does not impede transfection or promoter activity. The effect of BBR on reducing Htt aggregation was also shown by Western blotting that revealed aggregated Htt in the stacking gel. Quantification of the ratios of aggregated Htt to actin via densitometry also verified the reduction of Htt aggregates by BBR (Fig 1B and 1C) (for treatment with 50uM BBR, P = 0.028, T = 5.85, DF = 2). These results suggest that BBR can suppress the aggregate formation or the accumulation of mutant Htt (Fig 1A–1C).

Bottom Line: We found that BBR can reduce the accumulation of mutant huntingtin in cultured cells.We found that BBR could promote the degradation of mutant huntingtin by enhancing autophagic function.Since BBR is an orally-taken drug that has been safely used to treat a number of diseases, our findings suggest that BBR can be tested on different HD animal models and HD patients to further evaluate its therapeutic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Genetics, Emory University School of Medicine, 615 Michael Street, Atlanta, GA 30322, United States of America; Graduate Program of Microbiology and Molecular Genetics, Emory University, Atlanta, GA 30322, United States of America.

ABSTRACT
Huntington disease (HD) represents a family of neurodegenerative diseases that are caused by misfolded proteins. The misfolded proteins accumulate in the affected brain regions in an age-dependent manner to cause late-onset neurodegeneration. Transgenic mouse models expressing the HD protein, huntingtin, have been widely used to identify therapeutics that may retard disease progression. Here we report that Berberine (BBR), an organic small molecule isolated from plants, has protective effects on transgenic HD (N171-82Q) mice. We found that BBR can reduce the accumulation of mutant huntingtin in cultured cells. More importantly, when given orally, BBR could effectively alleviate motor dysfunction and prolong the survival of transgenic N171-82Q HD mice. We found that BBR could promote the degradation of mutant huntingtin by enhancing autophagic function. Since BBR is an orally-taken drug that has been safely used to treat a number of diseases, our findings suggest that BBR can be tested on different HD animal models and HD patients to further evaluate its therapeutic effects.

No MeSH data available.


Related in: MedlinePlus