艾司氯胺酮调控miR-204-5p/CXCR4轴减轻机械通气诱导的大鼠肺损伤

韦松里; 范舜钦; 邹敏; 黎斌

doi:10.16190/j.cnki.45-1211/r.2023.07.008

艾司氯胺酮调控miR-204-5p/CXCR4轴减轻机械通气诱导的大鼠肺损伤

Esketamine regulating miR-204-5p/CXCR4 axis to reduce mechanical ventilation-induced lung injury in rats

摘要

摘要: 目的：探讨艾司氯胺酮（Esket）调控miR-204-5p/趋化因子受体4（CXCR4）轴对机械通气诱导的大鼠肺损伤的影响。方法：将SD 大鼠随机分为对照组（NC 组）、模型组（model 组）、低剂量Esket 组（Esket-L 组）、高剂量Esket 组（Esket-H 组）、antagomir 阴性对照组（antagomir NC 组）、miR-204-5p antagomir 组、Esket-H+antagomir NC 组、Esket-H+miR-204-5p antagomir组。除NC 组外，其余各组大鼠机械通气4 h，在机械通气前3 d给予相应药物处理。比较各组肺组织湿重（W）/干重（D）比值、超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量及血清中白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）水平，苏木精—伊红（HE）染色观察大鼠肺组织病理变化，TUNEL 法检测肺组织细胞凋亡率，实时荧光定量PCR（RT-qPCR）法检测miR-204-5p 表达，western blotting法检测Bax、Caspase-3、CXCR4蛋白表达。结果：与model组比较，Esket-L组、Esket-H 组肺组织病理损伤减轻，肺损伤评分、W/D比值、细胞凋亡率、MDA、IL-6、TNF-α、Bax、Caspase-3、CXCR4表达水平降低，SOD活性和miR-204-5p表达量升高（均P＜0.05）。与model组、antagomir NC组比较，miR-204-5p antagomir组肺组织病理损伤加重，肺损伤评分、W/D比值、细胞凋亡率、MDA、IL-6、TNF-α、Bax、Caspase-3、CXCR4 表达水平升高，SOD 活性和miR-204-5p 表达降低（均P＜0.05）。miR-204-5p antagomir 减弱了Esket-H 对机械通气诱导的大鼠肺损伤的改善作用（P＜0.05）。双荧光素酶报告基因实验证实miR-204-5p 可靶向调控CXCR4 表达（P＜0.05）。结论：Esket 可能通过上调miR-204-5p 来抑制CXCR4 表达，进而减轻机械通气诱导的大鼠肺损伤。

Abstract: Objective: To explore the impact of Esketamine (Esket) on mechanical ventilation-induced lung injury in rats by regulating miR-204-5p/C-X-C motif receptor 4 (CXCR4) axis.Methods: SD rats were randomly grouped into control group (NC group), model group, low-dose Esket group (Esket-L group), high-dose Esket group (Esket-H group), antagomir NC group, miR-204-5p antagomir group, Esket-H+antagomir NC group, and Esket-H+miR-204-5p antagomir group.Except for the NC group, the rats in other groups were given mechanical ventilation for 4 hours, and the corresponding drugs were given 3 days before mechanical ventilation.The ratio of wet weight (W) to dry weight (D), the activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA) and the serum levels of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were compared among all groups.The pathological changes of lung tissues were observed by hematoxylin-eosin (HE) staining, the apoptotic rate of lung tissue cells was detected by TUNEL method, the expression of miR-204-5p was detected by realtime fluorescence quantitative polymerase chain reaction (RT-qPCR), and the expressions of Bax, Caspase-3 and CXCR4 were detected by western blotting.Results: Compared with the model group, pathological injury of lung tissue was reduced in Esket-L and Esket-H groups, lung injury score, W/D ratio, apoptosis rate, and expression levels of MDA, IL-6, TNF-α, Bax, Caspase-3 and CXCR4 decreased, and SOD activity and miR-204-5p expression increased (both P< 0.05); compared with the model group and the antagomir NC group, the pathological injury of lung tissue in the miR-204-5p antagomir group was aggravated, the lung injury score, W/D ratio, apoptosis rate, expression levels of MDA, IL-6, TNF-α, Bax, Caspase-3 and CXCR4 increased, and SOD activity and miR-204-5p expression decreased (both P< 0.05); miR-204-5p antagomir attenuated the improvement effect of high-dose Esket on mechanical ventilation-induced lung injury in rats (P< 0.05).Dual luciferase reporter assay confirmed that miR-204-5p could target the expression of CXCR4 (P< 0.05).Conclusion: Esket may inhibit the expression of CXCR4 by up-regulating miR-204-5p, thereby reducing mechanical ventilation-induced lung injury in rats.

HTML全文

参考文献(0)

施引文献

资源附件(0)