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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 increases autophagic activity in cultured cells.(A) Western blotting of LC3B in HEK293 cells transfected with or without Htt and treated with BBR (0, 5, 25, 50, or 100 μM). (B) Densitometry analysis of the ratios of LC3-I to LC3-II in above Western blots. (C) Western blotting of Htt-transfected HEK293 cells treated with or without 50 μM BBR or bafilomycin (BFA), an autophagy inhibitor. Antibody to P62, which decreases when autophagy activates, was used to confirm the altered activity of autophagy. (D) Densitometry analysis of the ratios of aggregated Htt or P62 to actin in Western blots in (C). The quantitative data are presented as mean±SE.
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pone.0134142.g002: BBR increases autophagic activity in cultured cells.(A) Western blotting of LC3B in HEK293 cells transfected with or without Htt and treated with BBR (0, 5, 25, 50, or 100 μM). (B) Densitometry analysis of the ratios of LC3-I to LC3-II in above Western blots. (C) Western blotting of Htt-transfected HEK293 cells treated with or without 50 μM BBR or bafilomycin (BFA), an autophagy inhibitor. Antibody to P62, which decreases when autophagy activates, was used to confirm the altered activity of autophagy. (D) Densitometry analysis of the ratios of aggregated Htt or P62 to actin in Western blots in (C). The quantitative data are presented as mean±SE.

Mentions: To determine autophagic flux, we examined LC3B, which converts from form I to from II to serve as the recruiter of autophagosome substrate P62 during the activation of autophagy. Non-transfected or Htt-transfected HEK293 cells were treated with 0, 5, 25, 50, or 100 μM BBR for 48 h. Western blotting analysis of LC3B-I and LC3B-II (Fig 2A) and densitometry were done to compare LC3-I/LC3-II ratio (Fig 2B). A lower ratio indicates more conversion of LC3-I to LC3-II or activation of autophagy. This ratio was decreased as the concentration of BBR was increased, suggesting that BBR could increase autophagic function (Fig 2B). Increased autophagic activity was also observed in both non-transfected cells and cells transfected with the control Htt-20Q (Fig 2B). To establish a causative relationship between autophagic increases and Htt aggregate reduction, we inhibited autophagy in Htt-120Q-transfected HEK293 cells using bafilomycin A (BFA) and treated these cells with 50 μM BBR. Western blotting showed that BBR reduced Htt aggregation and also antagonized the effect of BFA on increasing Htt aggregation (Fig 2C) (BBR only: P = 0.173, T = 2.08, DF = 2; BFA only: P = 0.124, T = 2.57, DF = 2). P62 was used as an autophagic indicator, as it is an expendable substrate that decreases with autophagic up-regulation. We also saw a decrease in P62 after BBR treatment and its increase after BFA treatment, verifying BFA’s activity to inhibit autophagy (BBR only: P = 0.008, T = 11.11, DF = 2; BFA only: P = 0.0008, T = 35.34, DF = 2). The opposite effects of BBR and BFA on Htt aggregation and P62 were further confirmed by densitometry quantifying of the Western blot results (Fig 2D).


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 increases autophagic activity in cultured cells.(A) Western blotting of LC3B in HEK293 cells transfected with or without Htt and treated with BBR (0, 5, 25, 50, or 100 μM). (B) Densitometry analysis of the ratios of LC3-I to LC3-II in above Western blots. (C) Western blotting of Htt-transfected HEK293 cells treated with or without 50 μM BBR or bafilomycin (BFA), an autophagy inhibitor. Antibody to P62, which decreases when autophagy activates, was used to confirm the altered activity of autophagy. (D) Densitometry analysis of the ratios of aggregated Htt or P62 to actin in Western blots in (C). The quantitative data are presented as mean±SE.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0134142.g002: BBR increases autophagic activity in cultured cells.(A) Western blotting of LC3B in HEK293 cells transfected with or without Htt and treated with BBR (0, 5, 25, 50, or 100 μM). (B) Densitometry analysis of the ratios of LC3-I to LC3-II in above Western blots. (C) Western blotting of Htt-transfected HEK293 cells treated with or without 50 μM BBR or bafilomycin (BFA), an autophagy inhibitor. Antibody to P62, which decreases when autophagy activates, was used to confirm the altered activity of autophagy. (D) Densitometry analysis of the ratios of aggregated Htt or P62 to actin in Western blots in (C). The quantitative data are presented as mean±SE.
Mentions: To determine autophagic flux, we examined LC3B, which converts from form I to from II to serve as the recruiter of autophagosome substrate P62 during the activation of autophagy. Non-transfected or Htt-transfected HEK293 cells were treated with 0, 5, 25, 50, or 100 μM BBR for 48 h. Western blotting analysis of LC3B-I and LC3B-II (Fig 2A) and densitometry were done to compare LC3-I/LC3-II ratio (Fig 2B). A lower ratio indicates more conversion of LC3-I to LC3-II or activation of autophagy. This ratio was decreased as the concentration of BBR was increased, suggesting that BBR could increase autophagic function (Fig 2B). Increased autophagic activity was also observed in both non-transfected cells and cells transfected with the control Htt-20Q (Fig 2B). To establish a causative relationship between autophagic increases and Htt aggregate reduction, we inhibited autophagy in Htt-120Q-transfected HEK293 cells using bafilomycin A (BFA) and treated these cells with 50 μM BBR. Western blotting showed that BBR reduced Htt aggregation and also antagonized the effect of BFA on increasing Htt aggregation (Fig 2C) (BBR only: P = 0.173, T = 2.08, DF = 2; BFA only: P = 0.124, T = 2.57, DF = 2). P62 was used as an autophagic indicator, as it is an expendable substrate that decreases with autophagic up-regulation. We also saw a decrease in P62 after BBR treatment and its increase after BFA treatment, verifying BFA’s activity to inhibit autophagy (BBR only: P = 0.008, T = 11.11, DF = 2; BFA only: P = 0.0008, T = 35.34, DF = 2). The opposite effects of BBR and BFA on Htt aggregation and P62 were further confirmed by densitometry quantifying of the Western blot results (Fig 2D).

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