摘要
目的探讨雷公藤红素对脂多糖(LPS)诱导脓毒症大鼠急性肺损伤(ALI)的影响及其机制。方法选择雄性SD大鼠18只,按随机数字表法随机分为对照组、模型组、雷公藤红素组,每组6只。模型组和雷公藤红素组经尾静脉一次性注射LPS 5 mg/kg建立ALI模型,雷公藤红素组同时经腹腔注射0.02%雷公藤红素1 mg/kg,对照组分别经尾静脉和腹腔注射等量生理盐水。注射完毕,常规饲养24 h,腹主动脉取血,采用血气分析仪检测腹主动脉血氧分压(PO2)。过量麻醉致死,无菌条件下解剖胸腔,取左肺组织,称湿重,烘干后称干重,计算肺组织湿重干重比值(W/D)。取右肺下叶组织,常规HE染色,光镜下观察肺组织病理改变,并行肺损伤病理评分。取右肺中上叶组织,冰浴下匀浆,离心留取上清液,采用ELISA法检测肺组织TNF-α、IL-1β、细胞间黏附分子1(ICAM-1)、血管内皮细胞黏附分子1(VCAM-1)含量。取右肺下叶组织,RIPA裂解液充分裂解,提取细胞质蛋白和细胞核蛋白,采用ELISA法检测细胞质和细胞核蛋白核因子κB(NF-κB)含量。结果与对照组比较,模型组和雷公藤红素组腹主动脉血PO2、肺组织细胞质内NF-κB含量均明显降低,肺组织W/D和肺组织TNF-α、IL-1β、ICAM-1、VCAM-1、细胞核内NF-κB含量以及肺组织病理评分均明显升高(P均<0.05);与模型组比较,雷公藤红素组腹主动脉血PO2、肺组织细胞质内NF-κB含量均明显升高,肺组织W/D和肺组织TNF-α、IL-1β、ICAM-1、VCAM-1、细胞核内NF-κB含量以及肺组织病理评分均明显降低(P均<0.05)。与对照组比较,模型组肺泡充血和出血明显,并伴有大量炎症细胞浸润,肺泡结构塌陷,轮廓不清;与模型组比较,雷公藤红素组肺泡充血和出血明显减轻,肺泡内可见少量炎症细胞浸润,肺泡结构清晰可见。结论雷公藤红素能够抑制LPS诱导的脓毒症大鼠ALI,其机制可能与抑制NF-κB的核内转位,从而抑制大鼠体内炎症因子含量有关。
Objective To investigate the effect and mechanism of celastrol on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in septic rats.Methods Eighteen male SD rats were randomly divided into the control group,model group,and celastrol group with 6 rats in each group.The rats in the model group were injected with 5 mg/kg LPS through tail vein to establish ALI models.The rats in the celastrol group were injected with the same amount of LPS through tail vein and 1 mg/kg of 0.02%celastrol intraperitoneally.The rats in the control group were separately injected with the same amount of normal saline through tail vein and abdominal cavity.After 24-hour culture,the abdominal aorta blood was collected,and PO2 of the abdominal aorta was detected by blood gas analyzer.Rats were executed with narcotic overdose,and the chests in the rats were dissected under aseptic conditions.The left lung tissues were taken,and the wet weight was weighed.After drying in the oven,the dry weight of rats was weighed,and the ratio of the wet weight to the dry weight of the lung tissues(W/D)was calculated.The inferior lobe of right lung in rats was taken and HE staining was per⁃formed to observe the pathological changes in the lung tissues under light microscope,and pathological score of lung injury was performed.The middle and superior lobes of right lung in the rats were homogenized in ice bath,and the supernatant was extracted by centrifugation.The content of TNF-α,IL-1β,intercellular adhesion molecule-1(ICAM-1),and vascular cell adhesion molecule-1(VCAM-1)in the lung tissues was determined by ELISA.The inferior lobe of right lung was tak⁃en and fully lysed with RIPA lysate.Cytoplasmic protein and nuclear protein were extracted according to the instructions of cytoplasmic nucleoprotein extraction kit.The content of NF-κB in the cytoplasm and nucleus was determined by ELISA.Results Compared with the control group,PO2 of abdominal aorta and cytoplasmic NF-κB in the lung tissues significant⁃ly decreased in the model group and celastrol group,while the W/D,the content of TNF-α,IL-1β,ICAM-1,and VCAM-1 in the lung tissues and NF-κB in nucleus,and pathological score of lung injury significantly increased(all P<0.05).Com⁃pared with the model group,PO2 of the abdominal aorta,NF-κB in cytoplasm of the lung tissues significantly increased in the celastrol group,while the W/D,the content of TNF-α,IL-1β,ICAM-1,and VCAM-1 in the lung tissues and NF-κB in nucleus,and pathological score of lung injury significantly decreased(all P<0.05).Compared with the control group,the alveolar hyperemia and bleeding were obvious in the model group,accompanied by a large number of inflammatory cell infiltration,and the alveolar structure collapsed and the contour was not clear.However,compared with the model group,the alveolar hyperemia and bleeding were significantly reduced in the celastrol group,and a small amount of inflammatory cell infiltration was observed in the alveolar,and the alveolar structure was clearly visible.Conclusion Celastrol can inhibit LPS-induced ALI in sepsis rats,and the mechanism may be related to the inhibition of nuclear translocation of NF-κB,thus inhibiting the content of inflammatory cytokines in rats.
作者
何招辉
杨春丽
杨小刚
黄翠兰
郭经华
HE Zhaohui;YANG Chunli;YANG Xiaogang;HUANG Cuilan;GUO Jinghua(Jiangxi Provincial People's Hospital,Nanchang 330006,China)
出处
《山东医药》
CAS
2021年第10期49-52,共4页
Shandong Medical Journal
基金
江西省卫生计生委中医药科研课题(2017A324)。
关键词
急性肺损伤
雷公藤红素
脂多糖
炎症反应
核因子ΚB
大鼠
acute lung injury
celastrol
lipopolysaccharide
inflammatory response
nuclear factorκB
rats
