心肌纤维化病理染色鉴定方法的综合评价

Comprehensive Assessment of Authenticating Method of Pathological Staining in Cardiac Fibrosis

目的观察比较苏木精-伊红染色法(hematoxylin and eosin staining,HE staining)、Masson染色法和苦味酸-天狼猩红染色法(picric sirius red staining,PSR staining)在评价心肌纤维化的各自特征及优缺点。方法 20只SD大鼠予以盐酸异丙肾上腺素(isoprenaline,ISO)制备心肌纤维化模型,以5mg/(kg·d)于同一时间段注射,持续3周。取大鼠心脏组织行HE染色、Masson染色以及PSR染色,置于显微镜下观察纤维化面积及胶原沉积特点,并比较Masson染色和PSR染色纤维化面积所占心肌总面积比重。结果 3种染色方法均可显示心肌纤维化面积,Masson染色和PSR染色所示纤维化面积差异无统计学意义(P>0.05);PSR染色后进一步行免疫荧光染色能够明确区分Ⅰ型和Ⅲ型胶原纤维的表达,然而Masson染色和HE染色则不能区分。结论 PSR染色法结合免疫荧光技术较Masson染色法和HE染色法,能够更好的评价心肌纤维化程度及胶原纤维类型,对于心肌纤维化的治疗及预防具有重要意义。

Objective To compare hematoxylin and eosin（HE） staining, Masson＇s staining and picric sirius red（PSR） staining in evaluation of cardiac fibrosis. Methods Twenty adult sprague dawley（SD） rats were given subcutaneous injection of isoprenaline for 5mg/（kg · d） for 3 weeks, which induced cardiac fibrosis model successfully. In order to observing the fibrotic areas and collagen deposi- tion, we prepared paraffin sections by using the heart specimen of rat, and carried out HE staining, Masson staining and PSR staining re- spectivelly for the special staining. Results All staining methods could clearly show the collagen fibers and fibrosis stage. We could clearly distinguish the type I and type 111 collagen fibers through PSR staining with immunoflurescent techniques, but not Masson and HE staining. Conclusion The results demonstrated that PSR staining is better than HE and Masson staining to be used for the evaluation of degree and type of proliferation of collagen fibers. It has important signification for treatment and prevention of cardiac fibrosis.