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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

Oral administration of BBR reduced mutant Htt aggregation and increased autophagy in transgenic N171-82Q mice.(A) Fluorescent immunohistostaining of mouse brain slices from transgenic N171-82Q HD mice and WT siblings, orally gavaged with 40 mg/kg BBR or vehicle daily. (B) Western blotting of the cortex samples of 3 mice per group (HD+BBR vs. HD+vehicle) showing mutant Htt aggregates in the stacking gel and P62. Non-specific immunoreactive bands are also shown on the blots. Actin was used to show the internal control protein on western blot. (C) Densitometry analysis of above Western blots. The data are mean±SE (n = 3). (D) Quantitative RT-PCR analysis of transgenic mutant Htt mRNA in the cortex of the BBR-treated or untreated N171-82Q mice. n = 3 mice per group. Data are presented as mean CT±SEM. (E) Western blotting of WT mouse cortex samples (3-mouse pool per group) after the mice had been orally gavaged with 40 mg/kg BBR or vehicle daily for 24 weeks and sacrificed 4 h after the last BBR administration. (F) Densitometry analysis of Western blots in (E) showing the ratio of P62 to actin. The quantitative data are presented as mean±SE.
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pone.0134142.g004: Oral administration of BBR reduced mutant Htt aggregation and increased autophagy in transgenic N171-82Q mice.(A) Fluorescent immunohistostaining of mouse brain slices from transgenic N171-82Q HD mice and WT siblings, orally gavaged with 40 mg/kg BBR or vehicle daily. (B) Western blotting of the cortex samples of 3 mice per group (HD+BBR vs. HD+vehicle) showing mutant Htt aggregates in the stacking gel and P62. Non-specific immunoreactive bands are also shown on the blots. Actin was used to show the internal control protein on western blot. (C) Densitometry analysis of above Western blots. The data are mean±SE (n = 3). (D) Quantitative RT-PCR analysis of transgenic mutant Htt mRNA in the cortex of the BBR-treated or untreated N171-82Q mice. n = 3 mice per group. Data are presented as mean CT±SEM. (E) Western blotting of WT mouse cortex samples (3-mouse pool per group) after the mice had been orally gavaged with 40 mg/kg BBR or vehicle daily for 24 weeks and sacrificed 4 h after the last BBR administration. (F) Densitometry analysis of Western blots in (E) showing the ratio of P62 to actin. The quantitative data are presented as mean±SE.

Mentions: Based on the results from cultured cells, we wanted to examine whether BBR also reduced mutant Htt accumulation and aggregation in the HD mouse brain. Immunofluorescent staining of cerebellar brain slices revealed a drastic reduction of mutant Htt aggregates in the BBR-treated HD mouse brains as compared to the untreated HD mouse brains (Fig 4A). Western blotting of brain cortex samples from 3 separate mice per group was done and quantified by densitometry, revealing nearly 50% reduction in aggregated Htt in the BBR-treated HD mice as compared to their untreated HD siblings (Fig 4B and 4C) (P = 0.03, T = 5.64, DF = 2). However, the antibody (mEM48) used is poorly suited for detecting soluble mutant Htt in animal tissue despite the presence of non-specific immunoreactive bands (Fig 4B). To investigate whether this reduction occurred at the transcriptional level, RT-qPCR was performed on the HD mouse brain samples that were treated with or without BBR. The results show that BBR did not alter the transcriptional levels of transgenic mutant Htt (Fig 4D) (P = 0.988, T = 0.017, DF = 2). Thus, BBR is likely to inhibit the accumulation of mutant Htt at the protein level. To examine whether BBR stimulates autophagy to clear mutant Htt, we also probed the western blots with an antibody to P62 and found a moderate decrease in P62 in the BBR-treated group (Fig 4B and 4C) (P = 0.06, T = 3.90, DF = 2). Because the BBR-treated HD mice were examined for their behaviors and then sacrificed when they were near death, the most pronounced effects of BBR on autophagy in the HD mouse brain might not have been detected. Thus, the WT control group, which was gavaged with BBR daily at the same time as their HD siblings, was given their last gavage and sacrificed 4 hours later. The brain cortex samples of BBR-treated and untreated mice were then collected for western blot analysis with anti-P62. Densitometry analysis revealed a much more marked decrease in P62 in the BBR-treated mouse brain (Fig 4E and 4F), which also supports the idea that BBR treatment can increase autophagic activity in the mouse brain.


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

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

Oral administration of BBR reduced mutant Htt aggregation and increased autophagy in transgenic N171-82Q mice.(A) Fluorescent immunohistostaining of mouse brain slices from transgenic N171-82Q HD mice and WT siblings, orally gavaged with 40 mg/kg BBR or vehicle daily. (B) Western blotting of the cortex samples of 3 mice per group (HD+BBR vs. HD+vehicle) showing mutant Htt aggregates in the stacking gel and P62. Non-specific immunoreactive bands are also shown on the blots. Actin was used to show the internal control protein on western blot. (C) Densitometry analysis of above Western blots. The data are mean±SE (n = 3). (D) Quantitative RT-PCR analysis of transgenic mutant Htt mRNA in the cortex of the BBR-treated or untreated N171-82Q mice. n = 3 mice per group. Data are presented as mean CT±SEM. (E) Western blotting of WT mouse cortex samples (3-mouse pool per group) after the mice had been orally gavaged with 40 mg/kg BBR or vehicle daily for 24 weeks and sacrificed 4 h after the last BBR administration. (F) Densitometry analysis of Western blots in (E) showing the ratio of P62 to actin. The quantitative data are presented as mean±SE.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4520448&req=5

pone.0134142.g004: Oral administration of BBR reduced mutant Htt aggregation and increased autophagy in transgenic N171-82Q mice.(A) Fluorescent immunohistostaining of mouse brain slices from transgenic N171-82Q HD mice and WT siblings, orally gavaged with 40 mg/kg BBR or vehicle daily. (B) Western blotting of the cortex samples of 3 mice per group (HD+BBR vs. HD+vehicle) showing mutant Htt aggregates in the stacking gel and P62. Non-specific immunoreactive bands are also shown on the blots. Actin was used to show the internal control protein on western blot. (C) Densitometry analysis of above Western blots. The data are mean±SE (n = 3). (D) Quantitative RT-PCR analysis of transgenic mutant Htt mRNA in the cortex of the BBR-treated or untreated N171-82Q mice. n = 3 mice per group. Data are presented as mean CT±SEM. (E) Western blotting of WT mouse cortex samples (3-mouse pool per group) after the mice had been orally gavaged with 40 mg/kg BBR or vehicle daily for 24 weeks and sacrificed 4 h after the last BBR administration. (F) Densitometry analysis of Western blots in (E) showing the ratio of P62 to actin. The quantitative data are presented as mean±SE.
Mentions: Based on the results from cultured cells, we wanted to examine whether BBR also reduced mutant Htt accumulation and aggregation in the HD mouse brain. Immunofluorescent staining of cerebellar brain slices revealed a drastic reduction of mutant Htt aggregates in the BBR-treated HD mouse brains as compared to the untreated HD mouse brains (Fig 4A). Western blotting of brain cortex samples from 3 separate mice per group was done and quantified by densitometry, revealing nearly 50% reduction in aggregated Htt in the BBR-treated HD mice as compared to their untreated HD siblings (Fig 4B and 4C) (P = 0.03, T = 5.64, DF = 2). However, the antibody (mEM48) used is poorly suited for detecting soluble mutant Htt in animal tissue despite the presence of non-specific immunoreactive bands (Fig 4B). To investigate whether this reduction occurred at the transcriptional level, RT-qPCR was performed on the HD mouse brain samples that were treated with or without BBR. The results show that BBR did not alter the transcriptional levels of transgenic mutant Htt (Fig 4D) (P = 0.988, T = 0.017, DF = 2). Thus, BBR is likely to inhibit the accumulation of mutant Htt at the protein level. To examine whether BBR stimulates autophagy to clear mutant Htt, we also probed the western blots with an antibody to P62 and found a moderate decrease in P62 in the BBR-treated group (Fig 4B and 4C) (P = 0.06, T = 3.90, DF = 2). Because the BBR-treated HD mice were examined for their behaviors and then sacrificed when they were near death, the most pronounced effects of BBR on autophagy in the HD mouse brain might not have been detected. Thus, the WT control group, which was gavaged with BBR daily at the same time as their HD siblings, was given their last gavage and sacrificed 4 hours later. The brain cortex samples of BBR-treated and untreated mice were then collected for western blot analysis with anti-P62. Densitometry analysis revealed a much more marked decrease in P62 in the BBR-treated mouse brain (Fig 4E and 4F), which also supports the idea that BBR treatment can increase autophagic activity in the mouse brain.

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