柚皮素对肺炎支原体感染小鼠肺组织修复效果及高迁移率族蛋白B1/核因子-κB信号通路的影响

Effect of naringenin on lung tissue repair and high mobility group protein B1/nuclear factor-κB signaling pathway in mice infected with Mycoplasma pneumoniae

目的探讨柚皮素对肺炎支原体(MP)感染小鼠肺组织的修复效果及高迁移率族蛋白B1(HMGB1)/核因子-κB(NF-κB)信号通路的影响。方法2022年5—9月,BALB/c小鼠建立MP感染模型,将建模成功的小鼠按照随机数表法分为模型组,柚皮素低(50 mg/kg)、高(100 mg/kg)剂量组,阿奇霉素(90 mg/kg)组。每组12只。另取12只小鼠作为对照组,各组给予对应干预2周。全自动血气分析仪检测小鼠动脉血氧分压(PaO2);称量小鼠肺湿质量,并计算肺系数;HE染色观察小鼠肺组织病理学变化并对肺损伤进行评分;酶联免疫吸附法检测小鼠肺组织中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、IL-8水平;荧光定量PCR法检测小鼠肺组织中HMGB1、NF-κB信使核糖核酸(mRNA)水平;蛋白质印迹法检测小鼠肺组织中HMGB1、NF-κB蛋白水平。结果对照组小鼠肺泡组织结构清晰正常;与对照组相比,模型组小鼠肺泡组织具有明显的炎症细胞浸润,肺泡壁变厚,红细胞、肺水肿液渗入,动脉血PaO2[(35.57±3.70)mmHg比(88.96±8.66)mmHg]显著降低(P<0.05),肺系数[(0.98±0.08)%比(0.51±0.04)%]、肺损伤评分[(3.63±0.34)分比(0.00±0.00)分]、肺组织中TNF-α[(40.07±5.04)ng/L比(11.43±1.56)ng/L]、IL-1β[(119.60±13.25)ng/L比(50.37±6.38)ng/L]、IL-8、HMGB1、NF-κB mRNA和蛋白水平显著升高(P<0.05);与模型组相比,柚皮素低、高剂量组、阿奇霉素组小鼠肺泡壁变薄,红细胞、分泌液减少,炎症细胞浸润减少,肺泡组织趋于正常,动脉血PaO_(2)降低不明显(P<0.05),肺系数、肺损伤评分、肺组织中TNF-α、IL-1β、IL-8、HMGB1、NF-κB mRNA和蛋白水平依次降低(P<0.05)。结论柚皮素能明显改善MP感染小鼠肺组织损伤,可能与抑制HMGB1/NF-κB信号通路的激活有关。

Objective To investigate the repair effect of naringenin on lung tissue and its effect on high mobility group protein B1(HMGB1)/nuclear factor-κB(NF-κB)signaling pathway in mice infected with Mycoplasma pneumoniae(MP).Methods From May 2022 to September 2022,MP infection model was established in BALB/c mice.The successful model mice were assigned into model group,naringenin low(50 mg/kg),high(100 mg/kg)dose group and azithromycin(90 mg/kg)group according to the random number method,with 12 mice in each group.Another 12 mice were selected as control group,and each group was given corresponding interven-tion for 2 weeks.The arterial partial pressure of oxygen(PaO2)was detected by automatic blood gas analyzer;the wet lung weight of mice was weighed,and the wet lung weight index was calculated;HE staining was used to observe the pathological changes of lung tis-sue in mice and score the lung injury;the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-8 in lung tissue of mice were detected by enzyme-linked immunosorbent assay;the messenger RNA(mRNA)levels of HMGB1 and NF-κB in lung tissue of mice were detected by fluorescence quantitative PCR;the protein levels of HMGB1 and NF-κB in lung tissue of mice were detected by Western blotting.Results The alveolar tissue structure of mice in control group was clear and normal;compared with the control group,the model group had obvious infiltration of inflammatory cells in alveolar tissue,thickening of alveolar wall,infiltration of red blood cells and pulmonary edema fluid,and a significant reduction in arterial blood PaO_(2)[(35.57±3.70)mmHg vs.(88.96±8.66)mmHg](P<0.05),and the lung wet weight index[(0.98±0.08)%vs.(0.51±0.04)%],lung injury score[(3.63±0.34)points vs.(0.00±0.00)points],TNF-α[(40.07±5.04)ng/L vs.(11.43±1.56)ng/L],IL-1β[(119.60±13.25)ng/L vs.(50.37±6.38)ng/L],IL-8,HMGB1,NF-κB mRNA and protein levels in lung tissue were significantly increased(P<0.05);compared with the model group,the mice in the naringenin low,high dose groups and azithromycin group had thinner alveolar walls,reduced red blood cells and secretion,reduced inflammatory cell infiltration,and tended to have normal alveolar tissue,and no obvious decrease in arterial blood PaO2(P<0.05),but the lung wet weight index,lung injury score,TNF-α,IL-1β,IL-8,HMGB1,NF-κB mRNA and protein levels in lung tissue were gradually decreased(P<0.05).Conclusion Naringenin can significantly improve lung tissue injury in mice infected with MP,which may be related to inhibit-ing the activation of HMGB1/NF-κB signaling pathway.