基于同步辐射衍射增强成像的CCl4诱导肝纤维化发展及逆转动态研究

Development and Reversal of Liver Fibrosis Induced by CCl4 Based on Synchrotron Radiation Diffraction-Enhanced Imaging

目的肝纤维化是慢性肝病向肝硬化进展的关键步骤,在组织学上是可逆的。本研究拟基于同步辐射衍射增强成像(DEI)评估肝纤维化发展及其逆转过程中显微影像特征的变化。材料与方法雄性BALB/c小鼠54只,腹腔注射CCl4诱导肝纤维化模型。于0 d(正常组),注射后8、14、21、42 d及停止注射后自发逆转8、18、35、45 d各取6只大鼠肝脏组织固定于中性福尔马林溶液中。在北京同步辐射装置4W1A形貌成像实验站对肝脏样品行DEI。利用灰度共生矩阵分析肝脏表面纹理变化。成像后行病理实验,基于天狼猩红染色评估肝纤维化胶原纤维沉积情况。结果DEI清晰地展示了肝纤维化样品表面粗糙程度的变化。基于灰度共生矩阵的纹理分析表明,从肝纤维化发展到逆转,逆差矩值先变小后变大,而惯量、差的均值及差的熵等先变大后减小。病理结果表明,与正常肝脏的(0.12±0.04)%相比,造模8~42 d的胶原面积比逐渐增大至(3.35±0.87)%,而逆转8~45 d过程中胶原面积比逐渐减少至(1.16±0.17)%。结论DEI清晰地展现了肝纤维化发展及逆转过程中肝脏表面纹理变化等病变情况,可以作为评估肝纤维化程度的一种新方法,有望在肝脏疾病的早期诊断和治疗中发挥重要作用。

Purpose Liver fibrosis is a key step in the progression of chronic liver disease to cirrhosis,which is reversible histologically.In this study,the changes in microscopic features during the development and reversal of liver fibrosis are assessed based on synchrotron radiation diffraction-enhanced imaging(DEI).Materials and Methods Fifty-four BALB/c male mice are enrolled to establish liver fibrosis model induced by intraperitoneal injection of CCl4.At day 0(normal group),8,14,21,42 days after injection and 8,18,35,45 days after spontaneous reversal,six liver tissues of rats were fixed in the medium formalin solution at each time point.DEI of liver samples was performed at 4W1A of Beijing synchrotron radiation facility.The textures of liver surface were analyzed by gray level co-occurrence matrix(GLCM).After imaging,pathological experiments were conducted to evaluate the collagen deposition of liver fibrosis based on the Sirius Red staining.Results The changes of the surface roughness of liver fibrosis samples were clearly showed by DEI.During liver fibrosis and its subsequent regression,inverse difference moment first decreased and then increased,while the inertia,difference average and difference entropy first increased and then decreased based on the GLCM.Pathological results showed that compared with normal liver(0.12±0.04)%,collagen area ratio gradually increased to(3.35±0.87)%in 8-42 days of modeling,and gradually decreased to(1.16±0.17)%in 8-45 days of reversal.Conclusion DEI clearly shows the changes of the surface texture during the development and reversal of liver fibrosis,indicating that DEI imaging can be used as a new method to evaluate the degree of liver fibrosis,and is expected to play an important role in the early diagnosis and treatment of liver diseases.