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

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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射线照射成功复制易/抗放射性肺纤维化动物模型,并制备放射性肺损伤相关组织芯片,C57BL/6J小鼠易发肺纤维化与其肺组织部分成纤维细胞活化及FN和LN的高表达相关。 OBJECTIVE: To establish the tissue microarray of C57BL / 6J and C3H / HeN-induced lung injury in rats and to study the changes and significance of the expression of FN, LN and a-SMA. Methods The animal models of C57BL / 6J and C3H / HeN were irradiated by 20Gy60Coγ ray. The content of hydroxyproline in the lung tissue was determined and the tissue microarray was prepared. Immunohistochemistry (IHC) staining and image analysis The changes of expression of fibronectin (FN), laminin (LN) and alpha-smooth muscle actin (a-SMA) in the process of radiation-induced lung injury were detected quantitatively. Results Tissue microarray was successfully prepared and its HE and immunohistochemical staining results were in good agreement with those of normal sections. 1 ~ 6mC57BL / 6J mice lung tissue lesions experienced inflammatory, proliferative and fibrosis stages, collagen deposition increased 1 ~ 3mFN after irradiation was significantly higher than the normal control group, 6m gradually decreased to normal after irradiation, LN expression The expression of a-SMA was stronger in C3H / HeN mice than that in C3H / HeN mice. The lung tissue of 1 ~ 6mC3H / HeN mice after irradiation showed the changes of interstitial inflammation, Compared with the control group, there was no significant change in LN expression at 1 ~ 3m after irradiation significantly increased, 6m decreased. Conclusions The animal model of easy / anti-radioactive lung fibrosis was successfully replicated by γ-ray irradiation, and the relevant tissue microarrays of radiation-induced lung injury were prepared. C57BL / 6J mice were prone to pulmonary fibrosis and some of them were activated by fibroblasts and high expression of FN and LN Related.
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