血必净注射液对体外循环后急性肺损伤大鼠肺泡Ⅱ型上皮细胞凋亡相关微RNA表达的影响

doi:10.20047/j.issn1673-7210.2022.31.05

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摘要目的研究血必净注射液对体外循环后急性肺损伤大鼠肺泡Ⅱ型上皮细胞凋亡相关微RNA（miRNA）表达的影响。方法采用生物信息学技术筛选与肺泡Ⅱ型上皮细胞凋亡相关的候选miRNA；30只雄性SD大鼠（20～22周龄，体重450～500 g）按随机数字表法将其分为假手术组、体外循环组和血必净组，假手术组不建立体外循环模型，体外循环组和血必净组建立体外循环模型。假手术组于血管穿刺前2 h、体外循环组于体外循环前2 h经腹腔注射4 ml/kg的生理盐水。血必净组于体外循环前2 h经腹腔注射血必净注射液4 ml/kg，体外循环结束后4 h放血处死大鼠，取肺组织标本，采用大鼠肺湿/干重比（W/D）和肺泡损伤数定量评价指标（IQA）评价肺损伤程度；rT-PCR观察血必净对体外循环后急性肺损伤肺泡Ⅱ型上皮细胞中候选miRNA表达的影响。结果通过生物信息学分析筛选得到与大鼠肺泡Ⅱ型上皮细胞凋亡相关的候选miRNAs 5条：rno-miR-17-5p、rno-miR-34a-5p、rno-miR-26a-5p、rno-miR-21a-5p和rno-miR-375-3p。与假手术组比较，体外循环组W/D和IQA升高，差异有统计学意义（P ＜ 0.05）。与体外循环组比较，血必净组W/D、IQA下降，差异有统计学意义（P ＜ 0.05）；与假手术组比较，体外循环组大鼠肺泡Ⅱ型上皮细胞miRNA-17-5p表达下调，而miRNA-34a-5p 表达上调，差异有统计学意义（P ＜ 0.05）；与体外循环组比较，血必净组肺泡Ⅱ型上皮细胞miRNA-17-5p表达上调，差异有统计学意义（P ＜ 0.05），而miR-34a-5p表达下调，差异无统计学意义（P ＞ 0.05），rno-miR-26a-5p、rno-miR-21a-5p和rno- miR- 375-3p在三组中差异无统计学意义（P ＞ 0.05）。结论血必净注射液可能通过上调miRNA-17-5p对体外循环后急性肺损伤起到治疗作用，初步揭示血必净注射液治疗体外循环后急性肺损伤的分子机制。

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	徐朝军1 张胜康1 张怡1 周代勇1 宋岚2

关键词 ：体外循环, 急性肺损伤, 血必净注射液, miRNA-17-5p, 生物信息学

Abstract：Objective To study the effect of Xuebijing Injection on the expression of apoptosis-related microRNA (miRNA) in alveolar typeⅡepithelial cells of cardiopulmonary bypass -induced acute lung injury rats. Methods The candidate miRNAs related to apoptosis of alveolar type Ⅱ epithelial cells were screened by bioinformatics technology; 30 male SD rats (20 to 22 weeks old, body weight 450 to 500 g) were divided into three groups according to the random number table method. Sham operation group, cardiopulmonary bypass group and Xuebijing group. In the sham operation group, 4 ml/kg of normal saline was injected intraperitoneally two hours before the arterial and venous puncture, and in the cardiopulmonary bypass group two hours before the cardiopulmonary bypass. In Xuebijing group, 4 ml/kg of Xuebijing Injection was injected intraperitoneally two hours before cardiopulmonary bypass, and the rats were sacrificed by bloodletting four hours after cardiopulmonary bypass, and lung tissue samples were taken. Lung wet/dry weight ratio (W/D) and alveolar injury quantitative assessment index (IQA) were used to evaluate the degree of lung injury. The effect of Xuebijing Injection on the expression of candidate miRNAs in alveolar type Ⅱ epithelial cells of cardiopulmonary bypass-induced acute lung injury was observed by RT-PCR. Results Five miRNAs related to apoptosis of alveolar type Ⅱ epithelial cells were screened by bioinformatics analysis rno-miR-17-5p, rno-miR-34a-5p, rno-miR-26a-5p, rno-miR-21a-5p, and rno-miR-375-3p. Compared with Sham operation group, W/D and IQA increased in cardiopulmonary bypass group, and the difference was statistically significant (P ＜ 0.05) Compared with the cardiopulmonary bypass group, the W/D and IQA of the Xuebiying group were significantly decreased, and the differences were statistically significant (P ＜ 0.05). The expression of miRNA-17-5p in alveolar typeⅡepithelial cells of cardiopulmonary bypass group was lower than that of sham operation group,and the difference was statistically significant (P ＜ 0.05), while the expression of miRNA-34a-5p was up-regulated (P ＜ 0.05); Compared with the cardiopulmonary bypass group, the expression of miRNA-17-5p in alveolar typeⅡepithelial cells in the XBJ group was up-regulated, and the difference was statistically significant(P ＞ 0.05), while the expression of miR-34a-5p was down- regulated, but there was no statistical difference between the two groups (P ＞ 0.05). Conclusion Xuebijing Injection may play a therapeutic role in the treatment of cardiopulmonary bypass-induced acute lung injury by up-regulating miRNA-17- 5p, and initially reveal the molecular mechanism of Xuebijing injection in the treatment of CPB-Induced acute lung injury.

Key words： Cardiopulmonary bypass Acute lung injury Xuebijing injection MicroRNA-17-5p Bioinformatics technology

基金资助:湖南省教育厅科学研究重点项目（20A364）；
湖南中医药大学中医学国内一流建设学科项目（湘教通2018- 469号）。

通讯作者: 宋岚（1975-），女，博士，副教授，硕士生导师；研究方向：中西医结合心肺保护的基础研究。

作者简介: 徐朝军（1974-），男，博士，主任医师，硕士生导师；研究方向：中西医结合心肺保护的临床与基础研究。

引用本文:

徐朝军1 张胜康1 张怡1 周代勇1 宋岚2. 血必净注射液对体外循环后急性肺损伤大鼠肺泡Ⅱ型上皮细胞凋亡相关微RNA表达的影响[J]. 中国医药导报, 2022, 19(31): 22-26,43.
XU Zhaojun1 ZHANG Shengkang1 ZHANG Yi1 ZHOU Daiyong1 SONG Lan2. Effect of Xuebijing Injection on expression of apoptosis-related microRNAs in alveolar type Ⅱ epithelial cells of cardiopulmonary bypass-induced acute lung injury rats. 中国医药导报, 2022, 19(31): 22-26,43.

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