红景天对急性呼吸窘迫综合征小鼠肺内皮细胞BMP9表达的影响  

Effects of Rhodiola rosea on the expression of BMP9 in pulmonary endothelial cells of acute respiratory distress syndrome mice

作　　者：蒋镥[1] 张卓一[1] 杨洞洞 郑兰芝[1] 蒋庆雨 JIANG Lu;ZHANG Zhuoyi;YANG Dongdong;ZHENG Lanzhi;JIANG Qingyu(Department of Emergency,The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310006,China)

机构地区：[1]浙江中医药大学附属第一医院急诊医学科,杭州310006

出　　处：《浙江中西医结合杂志》2024年第6期495-500,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2021ZB103);浙江中医药大学自然科学青年探索项目(No.2022JKZKTS28);浙江省医药卫生科技计划项目(No.2024KY1215)。

摘　　要：目的探讨红景天对肺血管通透性和骨形态发生蛋白9(BMP9)表达的影响。方法将90只C57BL/6小鼠按照随机数字表法分为假手术组、模型组和红景天低、中、高剂量组,每组18只。采用盲肠结扎穿孔法造模,假手术组开腹后关腹,红景天低、中、高剂量组分别用40、80、160 mg/kg红景天灌胃5 d,假手术组和模型组灌胃等量生理盐水。处死后,计算各组小鼠肺湿/干重比值,利用伊文思蓝染色法检测肺内皮细胞的渗透性,对小鼠肺组织进行免疫组化、Western blot法检测BMP9表达水平。使用脂多糖(LPS)诱导人脐静脉内皮细胞(HUVEC)形成内皮细胞炎性损伤模型,用CCK-8法检测细胞活性,筛选出合适的红景天给药浓度。将细胞分为对照组(无LPS诱导)、LPS组和25、50、100μg/mL红景天组,应用实时荧光定量PCR和Western blot检测BMP9表达情况。结果与模型组比较,红景天低、中、高剂量组小鼠肺湿/干重比值[(7.42±0.25)、(6.75±0.25)、(5.93±0.27)比(8.39±0.39),P<0.01]和肺内皮细胞的渗透性[(620.73±0.36)%、(469.26±0.39)%、(179.20±0.36)%比(706.86±0.37)%,P<0.05或P<0.01]均显著降低,且与红景天剂量呈正相关。Western blot和免疫组化检测发现,与假手术组比较,模型组BMP9表达明显减少;而红景天低、中、高剂量组BMP9表达较模型组增加,且与剂量呈正相关。实时荧光定量PCR结果显示,与对照组比较,LPS组HUVEC中BMP9 mRNA相对表达量显著下降[(0.18±0.08)比(1.00±0.11),P<0.01];与LPS组比较,红景天给药组BMP9 mRNA相对表达量随给药浓度增加而增加[(0.39±0.09)、(0.52±0.10)、(0.78±0.09)比(0.18±0.08),P<0.05或P<0.01],但均低于对照组。Western blot检测细胞BMP9蛋白表达发现,LPS诱导细胞炎性反应可以导致BMP9表达明显下降,红景天给药组的BMP9表达量均高于LPS组,且与剂量相关。结论红景天减轻了急性呼吸窘迫综合征小鼠肺损伤并在体外缓解了LPS诱导的内皮损伤,其�Objective To investigate the effects of Rhodiola rosea on pulmonary vascular permeability and bone morphogenetic protein 9(BMP9)expression.Methods Ninety C57BL/6 mice were divided into sham operation,model,and Rhodiola rosea low-,medium-and high-dose groups using the random number table method,with 18 mice in each group.The cecal ligation and perforation were used for modeling.The sham operation group underwent abdominal closure after laparotomy.Rhodiola rosea low-,medium-and high-dose groups were administrated with 40,80,160 mg/kg Rhodiola rosea by gavage for 5 days,respectively.Sham operation and model groups wereadministrated with an equal volume of normal saline by gavage.After euthanasia,the lung wet/dry weight ratio of mice in each group was calculated,and the permeability of lung endothelial cells was detected using Evans blue staining.The expression levels of BMP9 in the lung tissues of mice were detected using immunohistochemistry and Western blotting.Human umbilical vein endothelial cells(HUVEC)were induced by lipopolysaccharides(LPS)to establish an endothelial cell inflammatory injury model.The cell viability was detected using CCK-8 assay to determine the appropriate concentration of Rhodiola rosea.The cells were divided into control(without LPS induction),LPS,and 25,50,100μg/mL Rhodiola rosea groups.The expression levels of BMP9 were detected using quantitative real-time PCR and Western blotting.Results Compared with the model group,the lung wet/dry weight ratio in the low-,medium-and high-dose groups of Rhodiola rosea(7.42±0.25,6.75±0.25,5.93±0.27 vs.8.39±0.39,P<0.01)and pulmonary endothelial cell permeability[(620.73±0.36)%,(469.26±0.39)%,(179.20±0.36)%vs.(706.86±0.37)%,P<0.05,P<0.01]in mice were significantly decreased.Moreover,these reductions were positively correlated with the concentration of Rhodiola rosea.Western blotting and immunohistochemical analyses revealed a significant decrease in BMP9 expression in the model group compared to the sham operation group.However,BMP9 expression

关 键 词：小鼠 红景天 骨形态发生蛋白9 急性呼吸窘迫综合征 内皮细胞稳态 

分 类 号：R285.5[医药卫生—中药学]