Mechanism of action of EV-YF1 based on artificially synthesized Y RNA fragments from extracellular vesicles of cardiosphere-derived cells on improving myocardial ischemia-reperfusion injury in mice

doi:10.11958/20222017

Abstract

Abstract:

Objective To investigate the mechanism of extracellular vesicular Y RNA fragment (EV-YF1) in ameliorating myocardial ischemia/reperfusion injury (MIRI). Methods Eighteen C57 BL/6J mice were divided into the Sham group, the MIRI group and the EV-YF1 group according to random number table method, with six mice in each group. Myocardial ischemia-reperfusion model was established in the MIRI group and the EV-YF1 group. Ten minutes after reperfusion, the aorta was clipped in the EV-YF1 group, and 5 μg EV-YF1 was injected into the left ventricle. DharmaFECT4 without EV-YF1 was injected into the left ventricle of the MIRI group. Evans Blue/triphenyltetrazole chloride (TTC) double staining was used to detect myocardial infarction size 24 h after modeling. Myocardial histopathology was observed by HE staining. Interleukin (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) in peripheral blood were detected by enzyme-linked immunosorbent assay. The mRNA expression levels of IL-1β, IL-6 and TNF-α in myocardial tissue were detected by real-time quantitative PCR. The expression levels of B-lymphoblastoma -2 (Bcl-2), Bcl-2 related X protein (Bax) and Caspase-3 protein in myocardial tissue were detected by Western blot assay. Myocardial cell apoptosis was detected by TUNEL staining. Results Compared with the Sham group, the MIRI group showed obvious infarct areas, significant myocardial tissue damage, significant upregulation of IL-1β, IL-6 and TNF-α levels in peripheral blood. IL-1β, IL-6 and TNF-α mRNA expression in myocardial tissue were elevated. Bcl-2, Bax and Caspase-3 protein expression were increased (P < 0.05), and the number of TUNEL nuclear staining positive cells was significantly increased. Compared with the MIRI group, the ratio of infarct area was reduced after MIRI in the EV-YF1 group, myocardial injury was alleviated, serum levels of IL-1β, IL-6 and TNF-α were significantly decreased, and mRNA expression levels of IL-1β, IL-6 and TNF-α in myocardial tissue were decreased. Bcl-2 protein expression level was increased, while Bax and Caspase-3 protein expression levels were decreased (P<0.05). The number of TUNEL nuclear staining positive cells was decreased. Conclusion EV-YF1 can ameliorate myocardial ischemia-reperfusion injury in mice by reducing inflammation and inhibiting myocardial apoptosis.

Key words: extracellular vesicles, myocardial reperfusion injury, apoptosis, interleukin-6, tumor necrosis factor-alpha, EV-YF1

CLC Number:

R542.22

Figures/Tables 9

References 22

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基因名称	引物序列（5'→3'）	产物大小（bp）
IL-1β	上游：GAAGAAGAGCCCATCCTCTGT 下游：TGTTCACGGAGCCTGTAG	129
IL-6	上游：ACACTCTTCACACCCCTCTCCTTC 下游：GACCCTCACTCCTTCCCTTCTATCC	122
TNF-α	上游：CACCACGCTCTTCTGTCTACTGAAC 下游：CCATTAGCCCACTTCTTTCCCTCAC	147
GAPDH	上游：TGAGCAAGAGAGGCCCTATC 下游：AGGCCCCTCCTGTTATTATG	101

基因名称	引物序列（5'→3'）	产物大小（bp）
IL-1β	上游：GAAGAAGAGCCCATCCTCTGT 下游：TGTTCACGGAGCCTGTAG	129
IL-6	上游：ACACTCTTCACACCCCTCTCCTTC 下游：GACCCTCACTCCTTCCCTTCTATCC	122
TNF-α	上游：CACCACGCTCTTCTGTCTACTGAAC 下游：CCATTAGCCCACTTCTTTCCCTCAC	147
GAPDH	上游：TGAGCAAGAGAGGCCCTATC 下游：AGGCCCCTCCTGTTATTATG	101

组别	梗死面积比值	危险区域比值
Sham组	2.89±0.38	56.92±1.35
MIRI组	48.48±1.21^a	57.42±1.44
EV-YF1组	33.86±0.86^ab	58.59±0.67
F	693.900^＊＊	0.508

组别	梗死面积比值	危险区域比值
Sham组	2.89±0.38	56.92±1.35
MIRI组	48.48±1.21^a	57.42±1.44
EV-YF1组	33.86±0.86^ab	58.59±0.67
F	693.900^＊＊	0.508

组别	IL-1β	IL-6	TNF-α
Sham组	86.2±5.9	257.7±25.9	235.5±19.4
MIRI组	482.3±35.8^a	753.7±31.5^a	815.6±28.1^a
EV-YF1组	350.1±21.0^ab	494.8±25.8^ab	516.7±34.4^ab
F	69.480^＊＊	79.230^＊＊	107.600^＊＊