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. 2020 Nov;24(21):12272-12284.
doi: 10.1111/jcmm.15403. Epub 2020 Oct 6.

Host pre-conditioning improves human adipose-derived stem cell transplantation in ageing rats after myocardial infarction: Role of NLRP3 inflammasome

Tsung-Ming Lee  1   2 , Horng-Jyh Harn  3   4 , Tzyy-Wen Chiou  5 , Ming-Hsi Chuang  6   7 , Chun-Hung Chen  7 , Chi-Hsuan Chuang  8 , Po-Cheng Lin  7 , Shinn-Zong Lin  3   9
Affiliations

Host pre-conditioning improves human adipose-derived stem cell transplantation in ageing rats after myocardial infarction: Role of NLRP3 inflammasome

Tsung-Ming Lee et al. J Cell Mol Med. 2020 Nov.

Abstract

Functional decline of stem cell transplantation in ageing hosts is well documented. The mechanism for this is poorly understood, although it is known that advancing age does not provide an optimal milieu for exogenous stem cells to survive, engraft and differentiate. We showed that n-butylidenephthalide improved human adipose-derived stem cell (hADSC) engraftment via attenuating the production of reactive oxygen species (ROS). It remained unclear whether pre-treated hosts with n-butylidenephthalide can rejuvenate the ageing heart and improve hADSC engraftment by regulating the ROS/NLRP3 inflammasome-mediated cardiac fibrosis after myocardial infarction. One hour after coronary ligation, hADSCs were transplanted into the hearts of young and ageing Wistar rats that were pre-treated with or without n-butylidenephthalide for 3 days. At day 3 after infarction, myocardial infarction was associated with an increase in ROS levels and NLRP3 inflammasome activity with age. hADSC transplant effectively provided a significant decrease in ROS levels, NLRP3 inflammasome activity, IL-1β levels and cardiac fibrosis in either young or old infarcted rats. However, the beneficial effects of hADSCs were greater in young compared with old rats in terms of NLRP3 inflammasome activity. The infarcted ageing rats pre-conditioned by n-butylidenephthalide improved engraftment and differentiation of hADSCs and additionally attenuated cardiac fibrosis compared with hADSCs alone. The anti-inflammation effects of n-butylidenephthalide were reversed by SIN-1. In conclusions, the increased NLRP3 inflammasome activity plays the pathogenesis of ageing-related functional hADSC decline in the ageing hosts. n-butylidenephthalide-pre-treated ageing hosts reversibly ameliorate the harsh microenvironments, improve stem cell engraftment and attenuate cardiac fibrosis after myocardial infarction.

Keywords: NLRP3 inflammasome; adipose-derived stem cell; butylidenephthalide; myocardial fibrosis; reactive oxygen species.

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Conflict of interest statement

The authors confirm that there are no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
A, Identification of the transplanted hADSCs and their cardiac phenotype from the border zone 3 d after infarction. The grafted cells were identified, and their human nature was established by positive immunostaining with anti‐human mitochondrial antibodies (brown). Human stem cells were observed at implanted area of transplantation groups but not the vehicle. More grafted cells were identified in the infarcted group pre‐treated with BP compared with hADSCs alone. A, young; B, young/ADSC; C, old; D, old/ADSC; E, old(BP)/ADSC. B, Engraftment of hADSCs. qPCR for human Alu was performed. Alu‐DNA index (amount of Alu‐amplified DNA in hADSC recipient rats relative to that detected in young infarcted rats). The number of animals in each group is indicated in parentheses. *P < .05 compared with young/ADSC; † P < .05 compared with the infarcted group treated with old/ADSC
FIGURE 2
FIGURE 2
Experiment 1. A, Myocardial superoxide measured by lucigenin‐amplified chemiluminescence. B, DHE staining from the border zone 3 d after infarction (n = 5 each group). Experiment 2. C, Myocardial superoxide measured by lucigenin‐amplified chemiluminescence. The number of animals in each group is indicated in parentheses. *P < .05 compared with young; † P < .05 compared with the infarcted groups treated with young/ADSC and old; ‡ P < .05 compared with old/ADSC. § P < .05 compared with the infarcted groups treated with old and old/BP/SIN‐1
FIGURE 3
FIGURE 3
Experiment 1. The changes in NLRP3 inflammasome at day 3 from the border zone. The protein levels of NF‐κB (A), NLRP3 (B) and IL‐1β (C) assessed by Western blot were significantly increased in old than young rats after infarction. Relative abundance was obtained against that of β‐actin. To further confirm serum IL‐1β levels, ELISA was performed (D). The mRNA changes in NF‐κB (E), NLRP3 (F) and IL‐1β (G) were similar to those of proteins. Results are mean ± SD of 3 independent experiments (n = 5 each group). Experiment 2. Pre‐treated hosts with BP provided reduction of these cytokines, which can be reversed by adding SIN‐1 (H‐J). The number of animals in each group is indicated in parentheses. *P < .05 compared with young; † P < .05 compared with the infarcted groups treated with young/ADSC and old; ‡ P < .05 compared with old/ADSC; § P < .05 compared with the infarcted groups treated with old and old/BP/SIN‐1
FIGURE 4
FIGURE 4
Immunofluorescent staining for human mitochondria (green) and sarcomeric α‐actinin (red), and DAPI (blue). Identification of cardiac phenotype of the transplanted hADSCs 28 d after infarction. Cardiomyocyte phenotype of transplanted hADSCs was confirmed by coexpression of human mitochondria (green) and sarcomeric α‐actinin (cardiomyocyte phenotype, red). Visualization of nuclei with DAPI stain. In the merged image, coexpression of human mitochondria and sarcomeric α‐actinin showed co‐localization of antigen expression as indicated by yellow fluorescence. The number of animals in each group is indicated in parentheses. *P < .05 compared with infarcted rats treated with young/ADSC and old(BP)ADSC
FIGURE 5
FIGURE 5
Serial echocardiographic analysis of heart function. A‐E, M‐mode echocardiographic images obtained at 28 d after MI. A severe hypokinesia to akinesia in the interventricular (IVS) wall motion was detected. F, Fractional shortening as determined from 2D images at baseline before and then again at 3 and 28 d after surgery. The number of animals in each group is indicated in parentheses. *P < .05 compared with young at the same time point; † P < .05 compared with the infarcted group treated with young/ADSC at the same time point; ‡ P < .05 compared with old at the same time point; § P < .05 compared with old/ADSC at the same time point
FIGURE 6
FIGURE 6
Cardiac fibrosis at the remote zone at day 28 after infarction. A, Representative sections from the remote zone with Sirius Red staining (red, magnification 400×). The line length corresponds to 50 μm. The values are mean ± SD of 5 animals from each group. B, Hydroxyproline content was also shown to measure quantitative amount of fibrosis. The number of animals in each group is indicated in parentheses. *P < .05 compared with young; † P < .05 compared with the infarcted groups treated with young/ADSC and old; ‡ P < .05 compared with old/ADSC
FIGURE 7
FIGURE 7
Schematic representation illustrates the involvements of BP‐mediated NLRP3 inflammasomes in cardiac fibrosis in postinfarcted rats. Activation of NLRP3 inflammasome, composed of NLRP3, ASC and pro‐caspase‐1, is tightly regulated by two‐step signals. The first “priming” signal, such as MI‐mediated DAMP, enhances the expression of inflammasome components and target proteins via activation of transcription factor NF‐κB. The second “activation” signal activates NLRP3 which recruits the ASC scaffolding protein and procaspase‐1 allowing for homodimerization and autocatalytic activation of caspase‐1. Active caspase‐1 cleaves pro–IL‐1β into the active isoform. BP inhibits ROS production, which in turn suppresses two‐step signals of NLRP3 inflammasome activation. Inhibition of these signalling pathways by their respective inhibitors is indicated by the vertical lines. TLR, Toll‐like receptor; DAMPs, danger‐associated molecular patterns
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References

    1. Gould KE, Taffet GE, Michael LH, et al. Heart failure and greater infarct expansion in middle aged mice: a relevant model for postinfarction failure. Am J Physiol Heart Circ Physiol. 2002;282:H615‐H621. - PubMed
    1. Carlson BM, Faulkner JA. Muscle transplantation between young and old rats: age of host determines recovery. Am J Physiol. 1989;256(6 Pt 1):C1262‐C1266. - PubMed
    1. Smythe GM, Shavlakadze T, Roberts P, et al. Age influences the early events of skeletal muscle regeneration: studies of whole muscle grafts transplanted between young (8 weeks) and old (13–21 months) mice. Exp Gerontol. 2008;43:550‐562. - PubMed
    1. Mauro AG, Bonaventura A, Mezzaroma E, Quader M, Toldo S. NLRP3 inflammasome in acute myocardial infarction. J Cardiovasc Pharmacol. 2019;74:175‐187. - PubMed
    1. Schroder K, Tschopp J. The inflammasomes. Cell. 2010;140:821‐832. - PubMed

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