Efficacy and mechanism of hESC-MSCs in ameliorating post-cardiopulmonary resuscitation cardiac dysfunction in pigs

doi:10.3877/cma.j.issn.1674-6880.2025.05.001

Abstract

Abstract:

Objective

To investigate the effects and underlying mechanisms of human embryonic stem cell derived mesenchymal stem cells (hESC-MSCs) on post-resuscitation cardiac dysfunction in a porcine model of cardiac arrest (CA).

Methods

Twenty-seven healthy male white pigs were randomly allocated to three groups (9 pigs per group) using a random number table: a Sham group, a cardiopulmonary resuscitation (CPR) group, and an hESC-MSCs group. The Sham group only completed the preparations related to surgical catheter placement. The CA-CPR model was established by inducing ventricular fibrillation for 10 minutes via a right ventricular electrode, followed by 6 minutes of CPR in the CPR group and the hESC-MSCs group. At 5 minutes after successful resuscitation, the hESC-MSCs group received an intravenous infusion of 2.5 × 10⁶ cells/kg hESC-MSCs in 1 hour; the Sham and CPR groups received an equivalent volume of 0.9% NaCl solution. Cardiac function parameters, including stroke volume (SV) and global ejection fraction (GEF), were monitored using pulse indicator continuous cardiac output (PiCCO) at 2, 4, 8, 24, 48, and 72 hours after resuscitation, and serum concentrations of cardiac troponin I (cTnI) and creatine kinase MB (CKMB) were quantified via enzyme-linked immunosorbent assay (ELISA). At 24, 48, and 72 hours after resuscitation, three pigs per group were euthanized, and left ventricular myocardial tissue samples were collected. Myocardial apoptosis was assessed using TdT-mediated dUTP nick labeling (TUNEL). Expression levels of protein kinase R-like endoplasmic reticulum kinase (PERK), C/EBP homologous protein (CHOP), cysteinyl aspartate specific proteinase 12 (caspase 12), and caspase 3 were determined via western-blotting.

Results

Significant differences were observed in SV (F = 97.956, 54.970, 46.893, 8.685, 17.754, 18.070; all P < 0.05), GEF (F = 219.499, 95.886, 43.986, 15.809, 16.337, 8.050; all P < 0.05), serum cTnI (F = 112.367, 259.559, 83.680, 69.664, 55.522, 102.025; all P < 0.05), and serum CKMB (F = 77.607, 77.995, 64.601, 43.265, 47.544, 25.836; all P < 0.05) among the three groups at all post-resuscitation time points (2, 4, 8, 24, 48, and 72 hours). Compared with the CPR group, the hESC-MSCs group demonstrated significantly higher SV and GEF at all post-resuscitation time points (all P < 0.05). Significant differences were also observed in the myocardial apoptosis index (F = 68.585, 156.766, 189.283; all P < 0.001) and protein expression of PERK (F = 108.029, 33.089, 42.870; all P < 0.001), CHOP (F = 32.813, 34.275, 69.464; all P < 0.001), caspase 12 (F = 41.344, 16.836, 82.824; all P < 0.001), and caspase 3 (F = 74.913, 41.634, 43.085; all P < 0.001) among the three groups at 24, 48, and 72 hours after resuscitation. Compared with the CPR group, the hESC-MSCs group showed significantly reduced myocardial apoptosis index and downregulated expression of endoplasmic reticulum (ER) stress-related apoptotic proteins at the same time points (all P < 0.05).

Conclusion

Post-resuscitation treatment with hESC-MSCs ameliorates cardiac dysfunction in a porcine CA model, potentially via inhibition of ER stress-induced apoptotic pathways.

Key words: Cardiac arrest, Cardiopulmonary resuscitation, Cardiac dysfunction, Mesenchymal stem cells derived from human embryonic stem cells, Endoplasmic reticulum stress, Apoptosis, Pig

Jinjiang Zhu, Jiefeng Xu, Feng Ge, Guangli Cao, Xuguang Wang, Meiya Zhou, Tiejiang Chen, Mao Zhang. Efficacy and mechanism of hESC-MSCs in ameliorating post-cardiopulmonary resuscitation cardiac dysfunction in pigs[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2025, 18(05): 353-361.

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References 37

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组别	只数	体质量（kg）	心率（次/min）	平均动脉压（mmHg）	呼气末二氧化碳分压（mmHg）	pH值	乳酸（mmol/L）
Sham组	9	39.0 ± 1.7	83 ± 4	108 ± 8	38.7 ± 1.5	7.49 ± 0.03	1.2 ± 0.5
CPR组	9	38.4 ± 2.2	83 ± 6	103 ± 8	37.7 ± 0.7	7.51 ± 0.04	1.4 ± 0.4
hESC-MSCs组	9	39.0 ± 1.1	83 ± 5	103 ± 7	38.1 ± 1.8	7.52 ± 0.04	1.1 ± 0.4
F值		0.308	0.006	1.253	1.109	2.015	0.658
P值		0.738	0.994	0.304	0.346	0.155	0.527

组别	只数	体质量（kg）	心率（次/min）	平均动脉压（mmHg）	呼气末二氧化碳分压（mmHg）	pH值	乳酸（mmol/L）
Sham组	9	39.0 ± 1.7	83 ± 4	108 ± 8	38.7 ± 1.5	7.49 ± 0.03	1.2 ± 0.5
CPR组	9	38.4 ± 2.2	83 ± 6	103 ± 8	37.7 ± 0.7	7.51 ± 0.04	1.4 ± 0.4
hESC-MSCs组	9	39.0 ± 1.1	83 ± 5	103 ± 7	38.1 ± 1.8	7.52 ± 0.04	1.1 ± 0.4
F值		0.308	0.006	1.253	1.109	2.015	0.658
P值		0.738	0.994	0.304	0.346	0.155	0.527

组别	只数	复苏前	复苏后2 h	复苏后4 h	复苏后8 h	复苏后24 h	复苏后48 h	复苏后72 h
Sham组	9	46.7 ± 4.1	43.7 ± 3.7	43.7 ± 3.6	46.0 ± 3.4	41.0 ± 2.6	45.7 ± 1.2	42.0 ± 4.6
CPR组	9	46.0 ± 2.9	20.0 ± 3.2^a	22.6 ± 2.8^a	26.1 ± 3.9^a	27.7 ± 6.7^a	28.0 ± 4.4^a	26.3 ± 5.7^a
hESC-MSCs组	9	48.1 ± 2.6	27.6 ± 4.1^ab	33.7 ± 5.8^ab	38.6 ± 5.6^ab	44.0 ± 5.2^b	42.3 ± 4.9^b	48.0 ± 3.0^b
F值		0.995	97.956	54.970	46.893	8.685	17.754	18.070
P值		0.385	<0.001	<0.001	<0.001	0.017	0.003	0.003

组别	只数	复苏前	复苏后2 h	复苏后4 h	复苏后8 h	复苏后24 h	复苏后48 h	复苏后72 h
Sham组	9	46.7 ± 4.1	43.7 ± 3.7	43.7 ± 3.6	46.0 ± 3.4	41.0 ± 2.6	45.7 ± 1.2	42.0 ± 4.6
CPR组	9	46.0 ± 2.9	20.0 ± 3.2^a	22.6 ± 2.8^a	26.1 ± 3.9^a	27.7 ± 6.7^a	28.0 ± 4.4^a	26.3 ± 5.7^a
hESC-MSCs组	9	48.1 ± 2.6	27.6 ± 4.1^ab	33.7 ± 5.8^ab	38.6 ± 5.6^ab	44.0 ± 5.2^b	42.3 ± 4.9^b	48.0 ± 3.0^b
F值		0.995	97.956	54.970	46.893	8.685	17.754	18.070
P值		0.385	<0.001	<0.001	<0.001	0.017	0.003	0.003

组别	只数	复苏前	复苏后2 h	复苏后4 h	复苏后8 h	复苏后24 h	复苏后48 h	复苏后72 h
Sham组	9	36.8 ± 3.2	35.9 ± 1.9	36.7 ± 2.5	36.7 ± 3.1	37.0 ± 3.0	38.7 ± 2.3	35.3 ± 1.2
CPR组	9	38.1 ± 3.8	16.8 ± 1.7^a	18.9 ± 1.3^a	22.3 ± 1.9^a	24.7 ± 3.2^a	24.0 ± 3.0^a	26.0 ± 5.3^a
hESC-MSCs组	9	39.2 ± 4.6	22.8 ± 2.3^ab	26.6 ± 3.8^ab	29.4 ± 4.3^ab	37.3 ± 3.2^b	34.7 ± 4.2^b	35.7 ± 2.1^b
F值		0.874	219.499	95.886	43.986	15.809	16.337	8.050
P值		0.430	<0.001	<0.001	<0.001	0.004	0.004	0.003

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