炎调方对脓毒症急性肺损伤大鼠NF-κB信号通路影响的时效关系研究  

Study on time-effect relationship of impact of Yantiao-Prescription on NF-κB signaling pathway in rats with acute lung injury induced by sepsis

作　　者：何淼[1] 沈晓红[1] 熊旭东[1] 汪海慧[2] HE Miao;SHEN Xiaohong;XIONG Xudong;WANG Haihui(Shuguang Hospital Affiliated with Shanghai University of TCM,Shanghai 200021,China;Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海中医药大学附属曙光医院,上海200021 [2]上海市中医医院,上海200071

出　　处：《药物评价研究》2020年第12期2410-2415,共6页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81303105)。

摘　　要：目的验证炎调方对脓毒症急性肺损伤(ALI)大鼠的保护效应,并观察炎调方对核因子κB(NF-κB)信号通路主要指标活性调控的时效关系。方法清洁级健康雄性SD大鼠随机分为假手术组、模型组、炎调方(生药量为9.9 g/kg)组、地塞米松(0.45 mg/kg)组,均每天ig给药1次,连续给药3 d。假手术组、模型组予等体积生理盐水,末次给药2 h后进行手术。采用盲肠结扎穿孔术(CLP)制备脓毒症ALI模型,各组分别于造模后4、6、8、10、12、18、24 h进行HE染色后肺组织损伤程度评分、检测肺组织NF-κB/p65 mRNA表达量和血清NF-κB、肿瘤坏死因子(TNF-α)、白细胞介素(IL)-1β、IL-6、IL-8水平。结果与模型组比较,炎调方组造模后12、18、24 h肺组织损伤程度评分均显著降低(P<0.01)。炎调方组和地塞米松组肺组织NF-κB/p65 mRNA相对表达量、血清NF-κB水平均显著低于模型组(P<0.01);高于假手术组(P<0.01);炎调方组大鼠肺组织NF-κB/p65 mRNA相对表达量、血清NF-κB均随CLP后时间的延长呈现上升趋势,12 h后变化趋缓。不同时间点炎调方组和地塞米松组血清TNF-α、IL-1β、IL-6、IL-8均显著低于模型组(P<0.01),显著高于假手术组(P<0.01);炎调方组血清TNF-α、IL-1β、IL-6、IL-8随CLP后时间的延长呈现上升趋势。脓毒症ALI大鼠肺组织NF-κB/p65mRNA相对表达量与血清NF-κB和促炎因子TNF-α、IL-1β、IL-6、IL-8水平呈强正相关。结论炎调方对脓毒症ALI大鼠肺组织保护效应的机制与下调NF-κB基因表达、进而下调炎性信号通路下游的促炎细胞因子水平相关。Objective To verify the effect of Yantiao-Prescription protectingts on rats from sepsis-induced acute lung injury,and to investigate the time-effect relationship on the main observation indexes activity in NF-κB signaling pathway.Methods Healthy male SD rats of clean grade were randomly divided into sham operation group,model group,Yantiao-Prescription(crude drug dosage:9.9 g/kg)group and dexamethasone(0.45 mg/kg)group.All rats were ig once a day for three consecutive days.The sham operation group and model group were given equal volume of normal saline,and the operation was performed 2 h after the last administration.Acute lung injury model induced by sepsis was established in rats by CLP.At 4,6,8,10,12,18 and 24 h after CLP,10 rats of each group were sacrificed and lung tissue and serum samples were collected.The pathological changes of lung tissue were observed by HE staining and by grading injury degree.The expression of NF-κB/p65 mRNA in lung tissue was detected by real-time PCR.The levels of NF-κB,TNF-α,IL-1β,IL-6 and IL-8 in serum were detected by ELISA.Results Compared with the model group,the scores of lung tissue injury in the Yantiao-prescription group were significantly lower than those in the model group(P<0.01).The relative expression of NF-κB/p65 mRNA in lung tissue and serum NF-κB level in both groups were significantly lower than those in model group(P<0.01),and higher than that in sham operation group(P<0.01).The relative expression of NF-κB/p65 mRNA in lung tissue and serum NF-κB in Yantiao-Prescription group increased with the prolongation of CLP,and the change was slowed down after12 h.At different time points,serum TNF-α,IL-1β,IL-6,IL-8 in Yantiao-Prescription group and dexamethasone group were significantly lower than those in model group(P<0.01),and significantly higher than those in sham operation group(P<0.01);serum TNF-α,IL-1β,IL-6,IL-8 in Yantiao-Prescription group increased with the time after CLP.There was a strong positive correlation between the relative expression of

关 键 词：炎调方 脓毒症 急性肺损伤 时效关系 核因子-ΚB 促炎细胞因子 

分 类 号：R285.5[医药卫生—中药学]