甲基莲心碱通过调控NF-κB信号通路减轻脓毒症大鼠急性肺损伤
Neferine Attenuates Acute Lung Injury in Sepsis Rats by Regulating NF-κB Signaling Pathway
游燕 1闫智杰2
作者信息
- 1. 南昌大学第四附属医院药学部,江西南昌 330003
- 2. 南昌大学第四附属医院重症医学科,江西南昌 330003
- 折叠
摘要
[目的]探讨甲基莲心碱对脓毒症大鼠急性肺损伤的治疗作用及机制.[方法]采用盲肠结扎穿刺(GLP)法制备脓毒症大鼠模型,造模成功后,将大鼠分为模型组,甲基莲心碱低、高剂量组和甲基莲心碱高剂量+PMA[核转录因子κB(NF-κB)激活剂佛波酯]组,同时设置假手术组,每组15只.术后6 h,给予相应干预,连续2 d.采用苏木素-伊红(HE)染色观察肺组织形态学改变,并进行损伤病理评分;检测肺组织湿干质量比;血气分析仪检测动脉血氧分压(PaO2)和动脉血二氧化碳分压(PaCO2)水平;酶联免疫吸附法(ELISA)测定支气管肺泡灌洗液(BALF)中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)水平;Western Blot法检测肺组织中NF-κB p65、磷酸化NF-κB(p-NF-κB)p65(Ser536)、环氧合酶2(COX-2)蛋白表达水平.[结果]与假手术组比较,模型组大鼠肺组织损伤严重,肺组织损伤病理评分、湿干质量比及PaCO2水平显著升高(P＜0.01),PaO2水平显著降低(P＜0.01),BALF中TNF-α、IL-1β、IL-6水平以及肺组织中p-NF-κB p65/NF-κB p65比值、COX-2蛋白表达水平显著升高(P＜0.01);与模型组比较,甲基莲心碱低、高剂量组大鼠肺组织损伤明显改善,肺组织损伤病理评分、湿干质量比及PaCO2水平显著降低(P＜0.05或P＜0.01),PaO2水平显著升高(P＜0.05或P＜0.01),BALF中TNF-α、IL-1β、IL-6水平以及肺组织中p-NF-κB p65/NF-κB p65比值、COX-2蛋白表达水平显著降低(P＜0.05或P＜0.01).然而,PMA干预后,甲基莲心碱对脓毒症大鼠肺组织损伤的改善作用被明显逆转(P＜0.01).[结论]甲基莲心碱可改善大鼠脓毒症急性肺损伤及肺功能,其作用可能与抑制NF-κB信号通路激活,进而减少体内炎症因子释放有关.
Abstract
Objective To investigate the therapeutic effect of Neferine on acute lung injury in sepsis rats and its mechanism.Methods A rat sepsis model was prepared by cecal ligation and puncture(CLP)method.After successful modeling,the rats were divided into model group,neferine low-and high-dose groups,neferine high-dose+PMA[nuclear factor kappa B(NF-κB)activator phorbol ester]group,and sham operation group was set up,with 15 rats in each group.At 6 hours after operation,the corresponding intervention was given for 2 days.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of lung tissue,and the pathological damage score was performed.The wet-dry mass ratio of lung tissue was detected;the levels of arterial partial pressure of oxygen(PaCO2)and arterial partial pressure of carbon dioxide(PaCO2)were detected by blood gas analyzer.The levels of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β)and interleukin 6(IL-6)in bronchoalveolar lavage fluid(BALF)were measured by enzyme-linked immunosorbent assay(ELISA).The protein expression levels of NF-κB p65,phosphorylated NF-κB(p-NF-κB)p65(Ser536)and cyclooxygenase-2(COX-2)in lung tissue were detected by Western Blot.Results Compared with the sham operation group,the pathological injury of lung tissue in the model group was severe,the pathological injury score and wet-dry mass ratio of lung tissue as well as PaCO2 level were significantly increased(P＜0.01),PaO2 level was significantly decreased(P＜0.01),the levels of TNF-α,IL-1β and IL-6 in BALF,the ratio of p-NF-κB p65/NF-κB p65 and the protein expression level of COX-2 in lung tissue were significantly increased(P＜0.01).Compared with the model group,the pathological damage of lung tissue in the low-dose and high-dose neferine groups was significantly improved,the pathological damage score and wet-dry mass ratio of lung tissue as well as PaCO2 level were significantly decreased(P＜0.05 or P＜0.01),PaO2 level was significantly increased(P＜0.05 or P＜0.01),the levels of TNF-α,IL-1β and IL-6 in BALF,the ratio of p-NF-κB p65/NF-κB p65 and the protein expression level of COX-2 in lung tissue were significantly decreased(P＜0.05 or P＜0.01).However,after PMA intervention,the improvement of neferine on lung tissue injury in septic rats was significantly reversed(P＜0.01).Conclusion Neferine can improve acute lung injury and lung function in rats with sepsis,which may be related to inhibiting the activation of NF-κB signaling pathway and thereby reducing the release of inflammatory factors in vivo.
关键词
甲基莲心碱/脓毒症/急性肺损伤/NF-κB信号通路/炎症/大鼠Key words
neferine/sepsis/acute lung injury/NF-κB signaling pathway/inflammation/rats引用本文复制引用
基金项目
江西省卫生健康委科技计划项目(SKJP220211237)
出版年
2024
国家科技期刊平台
NSTL
万方数据