Exosomes derived from human umbilical cord mesenchymal stem cells promote myocardial repair after myocardial infarction under hypoxia

doi:10.3969/j.issn.2095-4344.1716

Abstract

Abstract:

BACKGROUND: Studies have shown that exosomes are an important mechanism of stem cell therapy for myocardial infarction. Hypoxic preconditioning affects the quantity and content of exosomes from human umbilical cord mesenchymal stem cells.
OBJECTIVE: To compare the effects of exosomes derived from human umbilical cord mesenchymal stem cells pretreated with hypoxia and cultured with normoxia on myocardial repair after myocardial infarction in rats, and to explore the possible mechanisms.
METHODS: (1) Human umbilical cord mesenchymal stem cells isolated from healthy newborn umbilical cord were cultured for fourth generations and identified. Passage 4 human umbilical cord mesenchymal stem cells with fusion degree of 50%-60% were pretreated for 48 hours in combination with serum-free medium and oxygen gradient culture (hypoxia and normoxia). Exosomes were then extracted from two sets of cell culture supernatants by ultracentrifugation. (2) Fifty Sprague-Dawley female rats provided by the Experimental Animal Center of Shanxi Medical University in China were enrolled, 40 of which were used to make animal models of myocardial infarction and the remaining 10 rats were subjected to sham operation. After successful modeling, 30 model rats were randomly assigned into model group, hypoxic exosomes group, normoxic exosomes group (n=10/group). One week after myocardial infarction, the heart was opened again, and the same volume of exosomes or PBS was injected at four points around the infarct area (the whitening area), 25 μL per point, 100 μL for each rat. One week after transplantation, echocardiography was performed to measure cardiac function. After completion of echocardiography, heart samples were taken and pathological sections were prepared for hematoxylin-eosin staining and Masson staining. Western blot assay was used to detect the expression of apoptosis-related proteins Cleaved-caspase 3 and Bcl-2.
RESULTS AND CONCLUSION: (1) One week after exosomes transplantation, the ejection fraction and fractional shortening rate of the left ventricle in hypoxic exosomes and normoxic exosomes groups significantly increased (P < 0.01), while left ventricular end-diastolic diameter and left ventricular end-systolic diameter significantly decreased (P < 0.01). The above indexes improved most significantly in the hypoxic exosomes group (P < 0.01). (2) Hematoxylin-eosin staining results showed that the cells were in alignment in the sham-operated group. Most serious lesions were found in the model group. The hypoxic exosomes group had less lesions than the normoxic exosomes group. (3) Masson staining results showed that the compared with the model group, the collagen volume fraction was decreased significantly in the two exosomes groups, especially in the hypoxic exosomes group (P < 0.01). (4) Western blot results showed that compared with the model group, the expression of Cleaved-caspase-3 decreased while the expression of Bcl-2 increased in the hypoxic and normoxic exosomes groups, and the variation in the protein expression was more obvious in the hypoxic exosomes group than the normoxic exosomes group (P < 0.01). To conclude, these findings reveal that exosomes derived from human umbilical cord mesenchymal stem cells under hypoxia can improve myocardial repair in Sprague-Dawley rats with myocardial infarction.

Key words: myocardial infarction, umbilical cord mesenchymal stem cells, hypoxic preconditioning, exosomes, mesenchymal stem cell transplantation, National Natural Science Foundation of China

CLC Number:

Zhang Pin, Guo Ying, Gao Yajie, Wang Zhendong, Li Baiyi, Zhang Xiaomin, Niu Yuhu, Liu Zhizhen, Ma Lihui, Niu Bo, Guo Rui. Exosomes derived from human umbilical cord mesenchymal stem cells promote myocardial repair after myocardial infarction under hypoxia[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(17): 2630-2636.

Figures/Tables 6

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