Abstract:

Objective　To explore the effect of apigenin on regulating the miR-152/bromodomain protein 4 (BRD4) axis and inhibiting high mobility group protein B1 (HMGB1) in the treatment of carbon monoxide (CO) induced brain injury. Methods　The mouse models of CO induced brain injury were established in February 2024, and were divided into a control group, a CO treatment group, an apigenin 25 mg/kg intervention group, and an apigenin 50 mg/kg intervention group by the random number table method, with 10 in each group. The control group was intraperitoneally given air, 170 ml/kg; the CO treatment group was given intraperitoneal injection of CO, 170 ml/kg; the apigenin 25 mg/kg intervention group established the CO models 24 h, and were intraperitoneally injected apigenin, 25 mg/kg, once per day; and the 50 mg/kg apigenin intervention group established the CO models 24 h, and were intraperitoneally injected apigenin, 50 mg/kg , once per day. The mice were euthanized by 3% isoflurane anesthesia after 90 minutes, 7 days, and 14 days, respectively. The brain tissue and peripheral blood were collected. The pathological and physiological conditions of the cortical and hippocampal regions of the brain tissue were compared between the two groups, and HE staining was used for morphological observation. Immunohistochemical detection of glial fibrillary acidic protein in the brain tissue was did; the serum concentrations of HMGB1, tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 in the mouse serum were detected by ELISA; Western blot/real time polymerase chain reaction was used to detect the expressions of BRD4 and HMGB1/mRNA in the cells; the flow cytometry was used to detect cell apoptosis. Analysis of variance was used. Results　The expressions of GFAP protein in the control group, the CO treatment group, the apigenin 25 mg/kg intervention group, and the apigenin 50 mg/kg intervention group were (1.11±0.06), (1.93±0.18), (1.32±0.09), and (1.09±0.05), respectively, with statistical differences between the groups (all P<0.05). The concentrations of TNF-α, IL-1β, and IL-6 in the CO treatment group were higher than those in the control group and the apigenin 50 mg/kg intervention group [(1.24±0.08) pg/L vs. (0.45±0.03) pg/L vs. (0.59±0.04) pg/L, (13.56±2.30) mg/L vs. (6.30±1.23) mg/L vs. (7.69±2.10)mg/L, and (3.86±0.22) pg/L vs. (0.14±0.01) pg/L vs. (1.35±0.06) pg/L], with statistical differences (all P<0.05). The levels of HMGB1/β-actin, TNF-α/β-actin, IL-1β/β-actin, and IL-6/β-actin in the CO treatment group were higher than those in the control group and the apigenin 50 mg/kg intervention group [(1.23±0.10) vs. (0.50±0.04) vs. (0.78±0.08), (0.61±0.06) vs. (0.24±0.02) vs. (0.37±0.02), (0.85±0.14) vs. (0.42±0.05) vs. (0.52±0.02), and (1.23±0.10) vs. (0.50±0.04) vs. (0.78±0.08)], with statistical differences (all P<0.05). The apoptosis rates of hippocampal neurons in the CO treatment group, the apigenin 25 mg/kg intervention group, and the apigenin 50 mg/kg intervention group were higher than that in the control group [(19.92±3.25)%, (15.63±2.25)%, and (11.23±3.55)% vs. (4.98±1.26)%], with statistical differences (all P<0.05). Conclusion　Apigenin inhibits the expression of HMGB1 by regulating the miR-152/BRD4 axis, and alleviates brain damage caused by carbon monoxide poisoning.

Key words:

Brain injury, Carbon monoxide poisoning, Apigenin, High mobility group protein B1

摘要：

目的　探讨芹菜素调节miR-152/溴结构域蛋白4（BRD4）轴抑制高迁移率族蛋白B1（HMGB1）治疗一氧化碳（CO）中毒脑损伤的研究。方法　本文采取实验型研究，于2024年2月建立小鼠CO中毒脑损伤模型，随机数字法将其分为对照组（10只）、CO处理组（10只）、芹菜素25 mg/kg干预组（10只）、芹菜素50 mg/kg干预组（10只）。对照组腹腔给予空气170 ml/kg；CO处理组给予CO腹腔注射170 ml/kg；芹菜素25 mg/kg干预组建立CO模型24 h，腹腔注射芹菜素25 mg/kg，1次/d；芹菜素50 mg/kg干预组建立CO模型24 h，腹腔注射芹菜素50 mg/kg，1次/d；分别在90 min、7 d及14 d后用3%异氟醚麻醉下对动物实施安乐死，收集脑组织以及外周血。对比两组脑组织皮质区以及海马区的病理生理情况，HE染色剂形态学观察；免疫组化检测脑组织胶质纤维酸性蛋白；酶联免疫吸附试验检测小鼠血清中的血清HMGB1、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）浓度及各组组织中的蛋白表达测定；蛋白质印迹法/实时定量聚合酶链反应检测细胞BRD4、HMGB1/mRNA表达；流式细胞术检测细胞凋亡。统计学方法采用方差分析。结果　正常对照组GFAP表达为（1.11±0.06）、CO处理组为（1.93±0.18）、芹菜素25 mg/kg干预组为（1.32±0.09）、芹菜素50 mg/kg干预组为（1.09±0.05），各组间比较差异有统计学意义（均P<0.05）。CO处理组TNF-α、IL-1β、IL-6含量均高于正常对照组、芹菜素50 mg/kg干预组［（1.24±0.08）pg/L比（0.45±0.03）pg/L比（0.59±0.04）pg/L、（13.56±2.30）mg/L比（6.30±1.23）mg/L比（7.69±2.10）mg/L、（3.86±0.22）pg/L比（0.14±0.01）pg/L比（1.35±0.06）pg/L］，差异均有统计学意义（均P<0.05）。CO处理组HMGB1/β-肌动蛋白（β-actin）、TNF-α/β-actin、IL-1β/β-actin、IL-6/β-actin水平含量均高于正常对照组、芹菜素50 mg/kg干预组［（1.23±0.10）比（0.50±0.04）比（0.78±0.08）、（0.61±0.06）比（0.24±0.02）比（0.37±0.02）、（0.85±0.14）比（0.42±0.05）比（0.52±0.02）、（0.87±0.06）比（0.38±0.04）比（0.48±0.04）］，差异均有统计学意义（均P<0.05）。CO处理组、芹菜素25 mg/kg干预组、芹菜素50 mg/kg干预组的海马神经细胞凋亡率均高于正常对照组［（19.92±3.25）%、（15.63±2.25）%、（11.23±3.55）%比（4.98±1.26）%］，差异均有统计学意义（均P<0.05）。结论　芹菜素通过调节miR-152/BRD4轴抑制HMGB1表达，从而减轻CO中毒脑损伤。

关键词:

脑损伤, 一氧化碳中毒, 芹菜素, 高迁移率族蛋白B1