养阴清肺方调控放射性肺炎大鼠外周血T细胞Notch1,Jagged1信号通路

Effect of Yangyin Qingfei Decoction on Regulating Notch1 and Jagged1 Signaling Pathway in Peripheral Blood T Cells of Rats with Radiation Pneumonia

目的:观察养阴清肺方调控放射性肺炎大鼠T细胞Notch信号受体1(Notch1)及Notch信号配体1(Jagged1)表达规律的影响,探讨其对放射性肺炎防治作用的机制。方法:将40只雌雄各半的SD大鼠以随机数字表法随机分为正常组、模型组、激素组、联合用药组,直线加速器对除正常组外的3组大鼠全胸单次照射,剂量为16 Gy。照射后第1天起各组予相应的药物灌胃。照射后4周,处死大鼠,取全肺称重计算肺系数,取大鼠肺组织进行苏木素-伊红(HE)染色,马松(Masson)染色,Notch1免疫组化染色,取全血标本进行免疫磁珠分选提取T细胞及流式细胞术检测Notch1比率,用提取的T细胞进行实时荧光定量PCR(Real-time PCR),蛋白免疫印迹法(Western blot)测定Notch1,Jagged1 mRNA及蛋白表达,观察各组大鼠在放射后4周肺系数改变及Notch1,Jagged1表达的变化。结果:放射后4周,各组肺系数差异明显,正常组与联合用药组肺系数最为接近;与模型组比较,养阴清肺方联合泼尼松龙可明显降低Notch1 T细胞所占比例(P<0.05),可明显降低Notch1,Jagged1mRNA表达(P<0.05),明显下调Notch1,Jagged1相关蛋白表达(P<0.05)。结论:养阴清肺方能明显下调放射性肺炎大鼠Notch1,Jagged1表达,可能是防治放射性肺炎发生发展的机制之一。

Objective： To observe the effect of Yangyin Qingfei decoction in regulating T cells Notch signal receptor 1（ Notch1） and Notch signal ligand 1（ Jagged1） in rats with radiation pneumonia,in order to explore its mechanism for preventing and treating radiation pneumonia. Method： Forty male and female SD rats were randomly divided into normal group,model group,hormone group and combination group. The dosage was16 Gy. On the first day after irradiation,the rats in each group were given the corresponding medicine. At the 4 th week after irradiation,the rats were put to death,and their lungs were weighed. The lung tissues were detected by hematoxylin-eosin（ HE） staining, Masson staining and Notch1 immunohistochemical staining.Immunohistochemistry was used to extract T cells,and flow cytometry was used to detect Notch1 percentage. The changes in lung function and the expressions of Notch1 and Jagged1 were observed 4 weeks later after irradiation.Result： At the 4 thweek after radiotherapy,there were significant differences in lung coefficient between groups,and the lung coefficient of normal group was the closest to combined treatment group. Compared with model group,Yangyin Qingfei decoction combined with prednisolone can significantly reduce the proportion of Notch1 T cells（ P 0. 05）,the expressions of Notch1,NICD and Jagged1 were down-regulated significantly（ P 0. 05）,and Notch1 and Jagged1 mRNA expressions were significantly decreased（ P 0. 05）. Conclusion： Yangyin Qingfei decoction can significantly reduce Notch1 and Jagged1 expressions of radioactive pneumonia rats,and may be one of prevention and treatment mechanisms for the occurrence and development of radiation pneumonia.