Čebašek Vita, Ribarič Samo
Cells Tissues Organs. 2016;201(3):211-9. doi: 10.1159/000444140. Epub 2016 Mar 30.
We have previously shown by 3D study that 2 weeks after nerve injury there was no change in the length of capillaries per muscle fibre length in rat extensor digitorum longus muscle (EDL). The primary goal of the present 2D study was to determine the capillarity of rat EDL 4 weeks after various modes of nerve injury. Additionally, we wished to calculate the same capillary/fibre parameters that were used in our 3D stereological study. EDL muscles derived from denervated (4 weeks after nerve injury), re-innervated (4 weeks after two successive nerve crushes) and age-matched controls from the beginning (CON-1) and the end (CON-2) of the experiment were analysed in two ways. Global indices of capillarity, such as capillary density (CD) and capillary/fibre (C/F) ratio, were determined by automatic analysis, local indices as the number (CAF) and the length of capillaries around individual muscle fibres (Lcap) in relation to muscle fibre size were estimated manually by tracing the muscle fibre outlines and the transversally and longitudinally cut segments of capillaries seen in 5-µm-thin muscle cross sections. Four weeks after both types of nerve injury, CD increased in comparison to the CON-2 group (p < 0.001) due to atrophied muscle fibres in denervated muscles and probably proliferation of capillaries in re-innervated ones. Higher C/F, CAF (both p < 0.001) and Lcap (p < 0.01) in re-innervated than denervated EDL confirmed this assumption. Calculated capillary/fibre parameters were comparable to our previous 3D study, which strengthens the practical value to the adapted 2D method used in this study.
我们之前通过三维研究表明,在大鼠趾长伸肌(EDL)中,神经损伤2周后,每根肌纤维长度的毛细血管长度没有变化。本二维研究的主要目的是确定在各种神经损伤模式下4周后大鼠EDL的毛细血管分布情况。此外,我们希望计算与我们的三维体视学研究中使用的相同的毛细血管/纤维参数。对实验开始时(CON-1)和结束时(CON-2)来自去神经支配(神经损伤4周后)、重新神经支配(连续两次神经挤压4周后)以及年龄匹配对照的EDL肌肉进行了两种方式的分析。通过自动分析确定毛细血管分布的整体指标,如毛细血管密度(CD)和毛细血管/纤维(C/F)比值,通过追踪5微米厚的肌肉横切面上可见的肌纤维轮廓以及毛细血管的横向和纵向切割段,手动估计局部指标,即与肌纤维大小相关的单个肌纤维周围的毛细血管数量(CAF)和毛细血管长度(Lcap)。两种神经损伤4周后,由于去神经支配肌肉中的肌纤维萎缩以及重新神经支配肌肉中毛细血管可能的增殖,CD与CON-2组相比增加(p < 0.001)。重新神经支配的EDL中较高的C/F、CAF(均p < 0.001)和Lcap(p < 0.01)证实了这一假设。计算得到的毛细血管/纤维参数与我们之前的三维研究结果相当这加强了本研究中所采用的二维方法的实用价值。