Effect of miR-146a and miR-155 on cardiac xenotransplantation

Zhao,Zhicheng; Qi,Feng; Liu,Tong; Fu,Weihua

doi:10.3892/etm.2016.3867

Effect of miR-146a and miR-155 on cardiac xenotransplantation

Authors:
- Zhicheng Zhao
- Feng Qi
- Tong Liu
- Weihua Fu
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Affiliations: Department of General Surgery, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China
Published online on: November 3, 2016 https://doi.org/10.3892/etm.2016.3867
Pages: 3972-3978
Copyright: © Zhao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the expression levels of miR-146a and miR-155 in a cardiac xenograft model treated with the immunosuppressant FK506, and to construct lentiviral vectors to further study the roles of miR-146a and miR‑155 in cardiac xenotransplantation. Expression levels of miR‑146a and miR‑155 were examined by quantitative polymerase chain reaction analysis and protein expression of RelA, which is a member of the nuclear factor‑κB family, was examined by western blot analysis. Pre‑miR‑146a and pre‑miR‑155 fragments were designed and synthesized according to MiRBase and were cloned into the plasmid pCDH1‑MCS1‑EF1‑copGFP. Recombinant plasmids were identified by enzyme digestion and sequencing. Survival time of cardiac grafts in the FK506 treatment group was significantly increased in comparison with the control group (P<0.05). In addition, the histopathological grading results were significantly decreased in the treatment group (P<0.05). A significant decrease in RelA protein expression levels was observed in the treatment group (P<0.05), along with a significant increase in miR‑146a expression levels (P<0.05) and a significant decrease in miR‑155 expression levels (P<0.05). Digestion and sequencing findings demonstrated that the insertion of miRNA into the plasmid pCDH1‑MCS1‑EF1‑copGFP conformed with the pre‑miRNAs, and the lentiviral vectors were concentrated to a titer of 5x107 IFU/ml. These findings demonstrated that FK506 is able to inhibit the rejection effect in a mouse‑to‑rat cardiac xenotransplantation model. FK506 treatment altered the expression levels of miR‑146a and miR‑155, indicating that they may have an important role in regulating the immune response to the rejection effect. miR‑146a and miR‑155 lentiviral vectors were successfully constructed for further experiments both in vitro and in vivo.

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