Notch1干扰对慢性阻塞性肺疾病大鼠肺组织中细胞凋亡的影响及机制研究  

Effect and its mechanism of Notch1 interference on apoptosis of lung tissue of rats with chronic obstructive pulmonary disease

作　　者：徐佳 丁凌[1] 王海燕[1] XU Jia;DING Ling;WANG Haiyan(Department of Respiratory Medicine,Hangzhou Third People's Hospital,Hangzhou 310009,China)

机构地区：[1]杭州市第三人民医院呼吸内科,310009

出　　处：《浙江医学》2024年第10期1017-1021,I0003,共6页Zhejiang Medical Journal

基　　金：杭州市科技计划引导项目(20211231Y040)。

摘　　要：目的探讨缺口受体-1(Notch1)干扰对慢性阻塞性肺疾病(COPD)大鼠肺组织中细胞凋亡及凋亡相关蛋白表达的影响。方法18只SPF级雄性大鼠按照随机数字表法分为对照组、模型组和干扰组,每组6只。制备COPD大鼠模型。造模前1天干扰组大鼠尾静脉注射1 mL Notch1干扰腺病毒(3×109 pfu/mL),对照组和模型组注射等体积0.9%氯化钠注射液。HE染色观察大鼠肺组织病理形态变化并测定半定量评分;测定肺血管壁厚度;Tunel染色检测肺组织中细胞凋亡情况;使用活性氧(ROS)检测试剂盒测定肺组织ROS水平;采用Western blot法测定肺组织Notch1、B淋巴细胞瘤-2(Bcl-2)和半胱氨酸蛋白酶(Caspase)-3蛋白表达。结果与模型组相比,干扰组大鼠肺组织半定量评分显著下降(P<0.01),管壁厚度占外径的百分比和管壁面积占血管总面积的百分比均显著降低(均P<0.01),Tunel染色阳性细胞凋亡率显著下降(P<0.01),肺组织中ROS水平显著降低(P<0.01),Notch1和Caspase-3蛋白表达灰度值均显著降低(均P<0.01),Bcl-2蛋白表达灰度值显著升高(P<0.01)。结论Notch1干扰可减少COPD大鼠肺组织的细胞凋亡,下调ROS水平,下调Notch1和Caspase-3蛋白表达及上调Bcl-2蛋白表达。Objective To explore the effect of Notch1 interference on the apoptosis of lung histiocyte and the expression of apoptosis-related protein in rats with chronic obstructive pulmonary disease(COPD).Methods All the 18 male rats with SPF grade were divided into a control group,a model group,and a Notch1 interference group according to ramdom table method,with 6 rats in each group.The COPD rat model was prepared.At the day before modeling,Notch1 interfering adenovirus was injected into the tail vein of rats in the Notch1 interfering group with 1 mL 3×109 pfu/mL adenovirus concentrate/piece,while the control and model groups were injected with 1 mL 0.9%sodium chloride injection,respectively.HE staining was used to observe the pathological changes of rat lung tissues and semi quantitative scoring was performed.The thickness of pulmonary vascular wall was measured.Tunel staining was used to detect apoptosis in lung tissues;reactive oxygen species(ROS)detection kit was used to measure ROS levels in lung tissues;Western blot was used to determine the expressions of Notch1,B lymphoblastoma-2(Bcl-2),and Caspase-3 proteins in lung tissues.Results Compared with the model group,the semi quantitative score of lung tissues in the Notch1 interference group of rats was significantly decreased(P<0.01),and the percentage of wall thickness to outer diameter(WT)and wall area to total vascular area(WA)were both significantly decreased(both P<0.01).The apoptosis rate of Tunel staining-positive cells,the ROS level in lung tissues,and the grayscale values of Notch1 and Caspase-3 protein expressions in the Notch1 interference group were all significantly decreased(all P<0.01),while the grayscale values of Bcl-2 protein expression were significantly increased(P<0.01).Conclusion Notch1 interference can reduce apoptosis in lung tissue of COPD rats,down-regulate ROS levels,and Notch1 and Caspase-3 protein expressions,and up-regulate Bcl-2 protein expression.

关 键 词：缺口受体-1干扰 慢性阻塞性肺疾病 细胞凋亡 

分 类 号：R563.9[医药卫生—呼吸系统]