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采用 RNA-seq 技术研究富含甲烷生理盐水处理后大鼠腹部皮瓣缺血再灌注损伤的转录组谱。

Investigation of transcriptome profile of ischemia/reperfusion injury of abdominal skin flaps in rats after methane-rich saline treatment using RNA-seq.

机构信息

Department of Plastic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.

Department of Plastic Surgery, Peking Union Medical College Hospital, Beijing, China.

出版信息

Clin Hemorheol Microcirc. 2021;78(2):127-138. doi: 10.3233/CH-201046.

DOI:10.3233/CH-201046
PMID:33554891
Abstract

OBJECTIVE

Here we examined the influence of methane-rich saline treatment (MS) on the whole transcriptome of the skin flaps during the ischemia/reperfusion (I/R) injuryusing RNA-sequence (RNA-seq).

METHODS

The rats were divided into three groups: the sham surgery group (SH),the I/R surgery group treated with physiological saline (I/R-P) or the I/R surgery group treated with the methane-rich saline (I/R-M) respectively. On the 72 hours after operation, the perfusion and the distribution of micro-circulatoryblood flow in skin flaps were observed by laser doppler flowmeters. The whole transcriptome alteration of the skin flaps was examined using RNA-seq. Moreover, the responses of the skin flaps to MRS treatment were examined using bio-informatic and q-PCR approaches after I/R injury.

RESULTS

The methane-rich saline (MS) treatment could expand survival area and improve the blood perfusion of the skin flaps after l/R injury. Compared to the I/R-P group, 474 genes significantly altered in the I/R-M group. These genes were mainly associated the development, the cell adhesion and migration. In addition, the PI3K-Akt signal pathway was meaningfully related to regulation of MS treatment. Q-PCR results confirmed that MS treatment positively regulated PI3K-Akt signal pathway relative genes and inhibited the cell adhesion relative genes.

CONCLUSION

These results proved that methane-rich saline may alleviate I/R injury and improve flap survival rate by regulating cell adhesion and PI3K-Akt signal pathway.

摘要

目的

本研究采用 RNA 测序(RNA-seq)技术,探讨富含甲烷的盐水处理(MS)对缺血再灌注(I/R)损伤皮肤瓣全转录组的影响。

方法

将大鼠分为三组:假手术组(SH)、生理盐水处理的 I/R 手术组(I/R-P)和富含甲烷的盐水处理的 I/R 手术组(I/R-M)。术后 72 小时,采用激光多普勒血流仪观察皮瓣的灌注和微循环血流分布。采用 RNA-seq 技术检测皮瓣全转录组的改变。此外,在 I/R 损伤后,采用生物信息学和 q-PCR 方法检测 MRS 处理对皮瓣的反应。

结果

富含甲烷的盐水(MS)处理可扩大 I/R 损伤后皮瓣的存活面积,改善皮瓣的血液灌注。与 I/R-P 组相比,I/R-M 组有 474 个基因显著改变。这些基因主要与发育、细胞黏附和迁移有关。此外,PI3K-Akt 信号通路与 MS 治疗的调节有显著的相关性。q-PCR 结果证实,MS 处理可正向调节 PI3K-Akt 信号通路相关基因,抑制细胞黏附相关基因。

结论

这些结果表明,富含甲烷的盐水可能通过调节细胞黏附和 PI3K-Akt 信号通路来减轻 I/R 损伤,提高皮瓣存活率。

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Investigation of transcriptome profile of ischemia/reperfusion injury of abdominal skin flaps in rats after methane-rich saline treatment using RNA-seq.采用 RNA-seq 技术研究富含甲烷生理盐水处理后大鼠腹部皮瓣缺血再灌注损伤的转录组谱。
Clin Hemorheol Microcirc. 2021;78(2):127-138. doi: 10.3233/CH-201046.
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Protective effect of hydrogen-rich saline on ischemia/reperfusion injury in rat skin flap.富氢生理盐水对大鼠皮瓣缺血再灌注损伤的保护作用。
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