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A PEG-based method for the isolation of urinary exosomes and its application in renal fibrosis diagnostics using cargo miR-29c and miR-21 analysis

  • Nephrology - Original Paper
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Abstract

Purpose

To assess a new and highly specific, but low-cost, easily performed and suitable for large-scale applications method for renal fibrosis (RF) diagnostics.

Methods

Thirty-five RF and twenty non-RF patients were enrolled in the study. An appropriate polyethylene glycol (PEG) was used to isolate urinary exosomes. The efficiency of isolation process was evaluated by the morphology and size observation, as well as the detection of specific markers (CD63, CD9). The expression level of exosomal miR-29c, miR-21 and the endogenous control snRNA-U6 were detected by qRT-PCR. The diagnostic potency of urinary exosomal miR-29c and miR-21 was estimated by the ROC method. Spearman’s rank-order correlations analysis was used to assess the correlation between the miRNAs and clinical parameters, including pathological index.

Results

PEG-based method for isolation urinary exosome was effective and could be completed with a relatively low-speed centrifugal machine. Exosomal miR-29c and miR-21 were detected in all samples. The analysis of miRNAs in urinary exosomes revealed significant dys-regulation of miR-29c and miR-21 associated with RF. Exosomal miR-29c and miR-21 could predict degree of RF with AUC of 0.8333 and 0.7639 (P < 0.05). Correlation analysis showed that the level of miR-29c had a significant negative relationship with eGFR and the interstitial relative area.

Conclusions

The PEG-based method for isolation urinary exosome is an inexpensive and easily performed approach. The application for cargo miRNA analysis is feasible. Urinary exosomal miR-29c may present a promising diagnostic approach.

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Acknowledgements

Supported by the School Foundation of Chengdu University of Traditional Chinese Medicine (Project No. CGPY1601) and key experimental technical items of Chengdu University of Traditional Chinese Medicine (SYZD2017002).

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Authors and Affiliations

  1. Clinical Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu, China

    Chun-yan Lv, Zi-yi Zhao, Jin-hui Li, Yan Chen & Jiao Lv

  2. Department of Fundamental Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, China

    Wei-jun Ding & Yi-li Wang

Authors
  1. Chun-yan Lv
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  2. Wei-jun Ding
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  3. Yi-li Wang
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  4. Zi-yi Zhao
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  5. Jin-hui Li
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  6. Yan Chen
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  7. Jiao Lv
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Correspondence to Chun-yan Lv.

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The authors declare that they have no competing interests.

Ethical standard

All studies were performed according to the ethical standards of the institutional and national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All the participators were informed of the study and signed the informed consent form.

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Informed consent was obtained from all individual participants included in the study.

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Lv, Cy., Ding, Wj., Wang, Yl. et al. A PEG-based method for the isolation of urinary exosomes and its application in renal fibrosis diagnostics using cargo miR-29c and miR-21 analysis. Int Urol Nephrol 50, 973–982 (2018). https://doi.org/10.1007/s11255-017-1779-4

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  • DOI: https://doi.org/10.1007/s11255-017-1779-4

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