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Alleviation of Lead-Induced Apoptosis by Puerarin via Inhibiting Mitochondrial Permeability Transition Pore Opening in Primary Cultures of Rat Proximal Tubular Cells

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Abstract

Previous study has demonstrated that mitochondrial-dependent apoptotic pathway is involved in the nephroprotective effect of puerarin (PU) against lead-induced cytotoxicity in primary cultures of rat proximal tubular (rPT) cells. To further clarify how PU exerts its antiapoptotic effects, this study was designed to investigate the role of mitochondrial permeability transition (MPT) and subsequent apoptotic events in the process of PU against Pb-induced cytotoxicity in rPT cells. The results showed that Pb-mediated mitochondrial permeability transition pore (MPTP) opening together with mitochondrial cytochrome c release, activations of caspase-9 and caspase-3, and subsequent poly-ADP-ribose polymerase (PARP) cleavage can be effectively blocked by the addition of PU. Simultaneously, upregulation and downregulation of Bcl-2 and Bax with increased Bcl-2/Bax ratio due to PU administration further alleviated Pb-induced mitochondrial apoptosis. Moreover, PU can reverse Pb-induced ATP depletion by restoring mitochondrial fragmentation to affect ATP production and by regulating expression levels of ANT-1 and ANT-2 to improve ATP transport. In summary, PU produced a significant protection against Pb-induced mitochondrial apoptosis in rPT cells by inhibiting MPTP opening to ameliorate the mitochondrial dysfunction.

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Abbreviations

MPT:

Mitochondrial permeability transition

rPT:

Rat proximal tubular

MPTP:

Mitochondrial permeability transition pore

ANT:

Adenine nucleotide translocase

cyt-c :

Cytochrome c

ROS:

Reactive oxygen species

BCA:

Bicinchonininc acid

MMP:

Mitochondrial membrane potential

PI:

Propidium iodide

CoCl2 :

Cobalt chloride

Calcein-AM:

Calcein acetoxymethyl ester

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Acknowledgments

This work was supported by the national nature science foundation of China (No. 31472251), a foundation for the author of national excellent doctoral dissertation of People’s Republic of China (No. 201266) and the fund of Fok Ying Tung education foundation under grant no. 141022.

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Daizong Road No. 61, Tai’an, 271018, China

    Zhong-Kun Wang, Xiang-Bin Song, Zhen-Yong Wang, Du-Bao Yang & Lin Wang

  2. Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, 611130, China

    Xue-Lei Zhou

  3. Institute of Pathogen Biology, Taishan Medical College, East Road of Yingsheng No.2, Tai’an, 271000, China

    Dong-Ming Zhuang

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  1. Zhong-Kun Wang
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  2. Xue-Lei Zhou
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  4. Dong-Ming Zhuang
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  5. Zhen-Yong Wang
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  6. Du-Bao Yang
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  7. Lin Wang
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Correspondence to Lin Wang.

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The authors declare that they have no conflicts of interest.

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Zhong-Kun Wang, Xue-Lei Zhou, and Xiang-Bin Song contributed equally to this work.

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Wang, ZK., Zhou, XL., Song, XB. et al. Alleviation of Lead-Induced Apoptosis by Puerarin via Inhibiting Mitochondrial Permeability Transition Pore Opening in Primary Cultures of Rat Proximal Tubular Cells. Biol Trace Elem Res 174, 166–176 (2016). https://doi.org/10.1007/s12011-016-0701-8

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