利用组织芯片技术研究C57BL/6J和C3H/HeN小鼠放射性肺损伤进程差异及其机制

目的建立C57BL/6J和C3H/HeN两种小鼠放射性肺损伤进程相关组织芯片,并应用其研究FN、LN和a-SMA的表达变化及意义。方法采用20Gy60Coγ射线照射C57BL/6J和C3H/HeN两种小鼠复制动物模型,测定肺组织中羟脯氨酸含量,制备组织芯片,应用免疫组织化学(im-munohistochemistry,IHC)染色方法与图像分析技术定量检测纤连蛋白(fibronectin,FN)、层粘连蛋白(laminin,LN)和a-平滑肌肌动蛋白(alpha-smooth muscle actin,a-SMA)在放射性肺损伤进程中的表达变化。结果组织芯片制备成功,其HE和免疫组织化学染色结果与普通切片具有良好一致性。照射后1~6mC57BL/6J小鼠肺组织病变经历炎症期、增生期和纤维化期,胶原沉积增多,照后1~3mFN表达明显高于正常对照组,照后6m逐渐减少至正常,LN表达在照射后呈渐进性增加,a-SMA表达强于C3H/HeN小鼠;照射后1~6mC3H/HeN小鼠肺组织主要表现为间质性炎症改变,FN表达于照射后1~6m与正常对照组相比无明显变化,LN表达于照射后1~3m明显增强,6m逐渐减少。结论γ射线照...

Studies on progression of radiation pulmonary injury in C57BL/6J and C3H/HeN mice and the underlying mechanism by yissue microarray

Objective To establish the tissue microarray for radiation pulmonary injury in C57BL/6J and C3H/HeN mice and apply this tissue microarray to study the role of FN, LN and a-SMA in the pathogenesis of radiation pulmonary fibrosis. Methods C57BL/6J and C3H/HeN mice were irradiated with 20 Gy 60Co gamma rays on the whole chest. The amount of hydroxyproline in lung was measured. Then the tissue microarray was prepared. The expressions of FN, LN and a-SMA in the lung were detected by immunohistochemistry (IHC) and image analysis. Results The tissue microarray was satisfactorily established, the results of HE and immunohistochemical staining using this tissue microarray were concordant with that of ordinary sections. The changes of the lungs in C57BL/6J mice were as follows: the pathological changes undergone interstitial pneumonia, proliferation and fibrosis. Significant increase of collagen deposition was detected. FN increased significantly on 1 and 3 months after irradiation (p<0.01), then decreased gradually to a normal level on 6 months after irradiation. LN elevated gradually after irradiation. The expressions of a-SMA were more intense than that in C3H/HeN mice. The changes of the lungs in C3H/HeN mice were as follows: the pathological changes mainly developed institial pneumonitis, no significant changes of FN expression were observed after irradiation compared to that in the control. LN increased significantly on 1 and 3 months after irradiation, and then decreased gradually. Conclusions The models of pulmonary fibrosis-prone and fibrosis-resistant were established by irradiated with gamma rays. The tissue microarray for radiation pulmonary injury was satisfactorily prepared. The fibrosis-sensitivity of C57BL/6J mice was related with the activation of fibroblasts and the high expression of FN and LN in lung.