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Preconditioned adipose-derived stem cells ameliorate cardiac fibrosis by regulating macrophage polarization in infarcted rat hearts through the PI3K/STAT3 pathway

Laboratory Investigation (2019) | Download Citation


Stem cells can modify macrophage phenotypes; however, the mechanisms remain unclear. We investigated whether n-butylidenephthalide (BP) primed adipose-derived stem cells (ADSCs) attenuated cardiac fibrosis via regulating macrophage phenotype by a PI3K/STAT3-dependent pathway in postinfarcted rats. Male Wistar rats after coronary ligation were allocated to receive either intramyocardial injection of vehicle, ADSCs (1 × 106 cells), BP-preconditioned ADSCs, (BP + lithium)-preconditioned ADSCs, (BP + LY294002)-preconditioned ADSCs, and (BP + S3I-201)-preconditioned ADSCs. ADSCs were primed for 16 h before implantation. BP-pretreated ADSCs increased the cell viability compared with naive ADSCs in the in vitro experiments. Infarct sizes were similar among the infarcted groups at the acute and chronic stages of infarction. At day 3 after infarction, post-infarction was associated with increased M1 macrophage infiltration, which was inhibited by administering naive ADSCs. Compared with naive ADSCs, BP-preconditioned ADSCs provided a significant increase of Akt and STAT3 phosphorylation, STAT3 activity, STAT3 nuclear translocation, myocardial IL-10 levels, and the percentage of M2 macrophage infiltration. The effects of BP on M2 polarization were reversed by LY294002 or S3I-201. Furthermore, the phosphorylation of both Akt and STAT3 was abolished by LY294002, whereas Akt phosphorylation was not affected following the inhibition of STAT3. The addition of lithium did not have additional effects compared with BP alone. After 4 weeks of implantation, ADSCs remained in the myocardium, and reduced fibrosis and improved cardiac function. BP-preconditioned ADSCs provided superior cardioprotection, greater ADSC engraftment, and antifibrotic effects compared with naive ADSCs. These results suggest that BP-pretreated ADSCs polarize macrophages into M2 cells more efficiently than naive ADSCs via the PI3K/STAT3 pathway.

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This work was supported by the grants of An-Nan Hospital (ANHRF107-04) and Ministry of Science and Technology (MOST-106-2314-B-039-049), Taiwan.

Author information


  1. Cardiovascular Institute, An Nan Hospital, China Medical University, Tainan, Taiwan

    • Tsung-Ming Lee
  2. Department of Medicine, China Medical University, Taichung, Taiwan

    • Tsung-Ming Lee
  3. Bioinnovation Center, Tzu Chi Foundation, Hualien, Taiwan

    • Horng-Jyh Harn
    •  & Shinn-Zong Lin
  4. Department of Pathology, Buddhist Tzu Chi General Hospital, Tzu Chi University, Hualien, Taiwan

    • Horng-Jyh Harn
  5. Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien, Taiwan

    • Tzyy-Wen Chiou
  6. Department of Technology Management, Chung Hua University, Hsinchu, Taiwan

    • Ming-Hsi Chuang
  7. Gwo Xi Stem Cell Applied Technology, Hsinchu, Taiwan

    • Ming-Hsi Chuang
    • , Chun-Hung Chen
    •  & Po-Cheng Lin
  8. Genomics Research Center, Academia Sinica, Taipei, Taiwan

    • Chi-Hsuan Chuang
  9. Department of Neurosurgery, Buddhist Tzu Chi General Hospital, Tzu Chi University, Hualien, Taiwan

    • Shinn-Zong Lin


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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval

All rats had unrestricted access to food/water in accordance with the China Medical University Committee on Animal Care (Permit Number: 2016-070). The investigation conforms to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996).

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Correspondence to Shinn-Zong Lin.

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