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Berberine Induces Cell Apoptosis through Cytochrome C/Apoptotic Protease-Activating Factor 1/Caspase-3 and Apoptosis Inducing Factor Pathway in Mouse Insulinoma Cells

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Abstract

Objective

To investigate apoptotic effects of berberine, a significant alkaloids component existing in Rhizoma coptidis, and its possible acting mechanism in insulinoma cells.

Methods

Different concentrations of berberine were used to treat mouse insulinoma (MIN6) cells for various period of time. The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay, flow cytometry and enzyme-linked immuno sorbent assay. Changes in the relating pro- and anti-apoptosis proteins were detected by western-blotting.

Results

The half-maximal inhibitory concentration (IC50) of berberine was 5.7 μmol/L on MIN6 cells viability for 16 h. Berberine caused a 20% reduction (P<0.05) in cell number after only 4-h incubation; which reached 50% after 24 h (P<0.01). Berberine treatment for 16 h significantly increased the level of DNA fragmentation. The flow cytometry showed the apoptotic rate increased 2.9- and 4.6-fold after treating with berberine (5 μmol/L) for 8 and 16 h, while 3- and 8.7-fold after 10 μmol/L treatment for 8 and 16 h (P<0.01). Berberine treatment dramatically elevated the expression ratio of Bax to Bcl-2. Meanwhile, berberine notably increased the apoptosis-inducing factors and cytochrome C transforming from the mitochondria to the cytoplasm. Apoptotic protease-activating factor 1 (Apaf-1) was subsequently activated after cytochrome C release. Furthermore, caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.

Conclusion

High concentration (5 and 10 μmol/L) of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor (AIF) pathway.

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Authors and Affiliations

  1. School of Basic Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China

    Xin Fang, Qian-qian Mu, Na Yu & Si-hua Gao

  2. Fraser Laboratories for Diabetes Research, Department of Medicine, RI-McGill University Health Centre, Montreal, Quebec, H4A3J1, Canada

    Xin Fang, Xiao-liang Miao & Jun-li Liu

  3. School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, China

    Xiao-liang Miao, Min Wang & Hong-ping Yin

  4. Diabetes Research Center, Beijing University of Chinese Medicine, Beijing, 100029, China

    Dong-wei Zhang, Dan-dan Zhao, Fang-fang Mo & Si-hua Gao

Authors
  1. Xin Fang
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  2. Xiao-liang Miao
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  3. Jun-li Liu
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  4. Dong-wei Zhang
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  5. Min Wang
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  7. Qian-qian Mu
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  8. Na Yu
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  9. Fang-fang Mo
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  10. Hong-ping Yin
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  11. Si-hua Gao
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Contributions

The conception and study design: Gao SH, Liu JL, Yin HP; acquisition of data, analysis and interpretation of data: Fang X, Miao XL, Mu QQ, Yu N; drafting the article and revising it critically for important intellectual content: Fang X, Miao XL, Zhang DW, Zhao DD, Wang M, Mo FF.

Corresponding author

Correspondence to Si-hua Gao.

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Conflict of Interest

The authors do not have any conflict of interest to declare.

Supported by the National Natural Science Foundation of China (No. NSFC81274041 and No. NSFC81273995), the International Cooperation Projects of Ministry of Education (No. 2011DFA30920), the Key Drug Development Program of Ministry of Science an Technology (No. 20122X09103201-005)

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Fang, X., Miao, Xl., Liu, Jl. et al. Berberine Induces Cell Apoptosis through Cytochrome C/Apoptotic Protease-Activating Factor 1/Caspase-3 and Apoptosis Inducing Factor Pathway in Mouse Insulinoma Cells. Chin. J. Integr. Med. 25, 853–860 (2019). https://doi.org/10.1007/s11655-015-2280-5

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