【摘要】 目的 检测大鼠在弥漫性脑损伤（DBI）后海马区A1/A2型星形胶质细胞（A1s、A2s）随时间的表达变化。方法 选取SD雄性大鼠48只，随机分为对照组和DBI后4 h、8 h、12 h、1 d、3 d、5 d、7 d组，每组6只，通过改良的Marmarou脑损伤打击装置建立大鼠脑损伤模型，通过大体观察、苏木素-伊红（HE）染色观察大鼠脑组织损伤情况，通过免疫组织化学染色、实时荧光定量聚合酶链式反应（PCR）检测各组大鼠海马区A1s、A2s标志物C3d、S100A10的蛋白及mRNA的表达变化。结果 大体观察显示，对照组大鼠未见明显异常；实验组大鼠可见局限性蛛网膜下腔出血，侧脑室和脑组织腹侧面少量出血，脑组织可见轻度水肿，未见局灶性脑挫伤及脑挫裂伤。HE染色结果显示，实验组大鼠DBI后的大脑皮层及深部脑组织形态学改变，大鼠颅脑损伤为弥漫性，DBI造模成功。免疫组织化学染色结果显示，对照组大鼠海马区C3d蛋白的平均光密度值为（0.002 6±0.000 3）,DBI后4 h升高至（0.003 4±0.000 1），于DBI后1 d达高峰，为（0.015 8±0.000 4），染色程度加深，随后逐渐下降，7 d时为（0.003 6±0.000 2）;DBI后8 h、12 h、1 d、3 d、5 d组大鼠的海马组织C3d蛋白的平均光密度值高于对照组，差异均有统计学意义（均P<0.05）。对照组大鼠海马区S100A10蛋白的平均光密度值为（0.078 7±0.006 8）,DBI后4 h下降至（0.051 9±0.002 1）,1 d时最少，为（0.005 2±0.000 2），随后上升，7 d时为（0.034 6±0.001 8）;DBI后4 h、8 h、12 h、1 d、3 d、5 d、7 d组大鼠的海马组织S100A10蛋白的平均光密度值低于对照组，差异均有统计学意义（均P<0.05）。实时荧光定量PCR检测结果显示，C3d于DBI后4 h出现升高的趋势，8 h左右达高峰，表达量为（9.365 2±0.543 8）2^-Δ△Ct，随后逐渐下降；S100A10在DBI 4 h后开始下降，8 h左右表达量最低，表达量为（0.446 9±0.007 8）2^-Δ△Ct，随后呈上升趋势。DBI后4 h、8 h、12 h、1 d、3 d、5 d、7 d组大鼠的C3d mRNA表达量均高于对照组，DBI后4 h、8 h、3 d、5 d、7 d组大鼠的S100A10 mRNA表达量均低于对照组，差异均有统计学意义（均P<0.05）。结论 DBI后7 d内A1s、A2s呈单峰表达的时序性变化规律，且趋势相反，为DBI时间推断、临床用药及治疗提供一定的理论依据。更多还原

【Abstract】 Objective To investigate the expression changes of A1/A2 astrocytes over time after diffuse brain injury （DBI） in the hippocampus of mice.Methods A total of 48 male SD mice were randomly divided into normal control group and 4 h,8 h,12 h,1 d,3 d,5 d,7 d group after DBI （n=6）.The DBI model was established by modified Marmarou brain injury strike device.Brain injury in mice was observed by gross observation and hematoxylin-eosin （HE） staining.The expression of C3d,S100A10 and their mRNA,the markers of A1/A2 astrocytes （A1s/A2s） in the hippocampus of mice were detected by immunohistochemical staining and real-time quantitative Polyerase Chain Reaction （PCR）.Results Gross observation showed that there was no obvious abnormality in the normal control group,but there were localized subarachnoid hemorrhage,a small amount of hemorrhage in the lateral ventricle and ventral side of brain tissue,mild edema and no focal brain contusion and laceration in the experimental group.The results of HE staining showed that the morphological changes of cerebral cortex and deep brain tissue in the experimental group after DBI,and the craniocerebral injury of mice was diffuse.DBI model was successfully established.The results of immunohistochemical staining showed that the mean density of C3d protein in the hippocampus of the normal control groupmice was（0.002 6±0.000 3）.It increased to （0.003 4±0.000 1） at 4 h after DBI,and reached a peak of （0.015 8±0.000 4） at about 1 d after DBI.The staining was significantly deepened,and then the mean density gradually decreased.It was still higher than that in the normal control group at 7 d after DBI,the mean density was （0.003 6±0.000 2）.The mean density of C3d protein in the hippocampus of 8 h,12 h,1 d,3 d and 5 d after DBI were all higher than that in the normal control group,and the differences were statistically significant （P<0.05）.The mean density of S100A10 protein in the hippocampus of the normal control group mice was （0.078 7±0.006 8）,which decreased to （0.051 9±0.002 1） at 4 h after DBI,and was the lowest at 1d,reaching （0.005 2±0.000 2）,then showing an upward trend,（0.034 6±0.001 8） at 7 d.The mean density of S100A10 protein in hippocampus of mice in groups 4 h,8 h,12 h,1 d,3 d,5 d and 7 d after DBI was lower than that in the normal control group,and the difference was statistically significant （P<0.05）.The results of real-time fluorescent quantitative PCR showed that C3d increased at 4 h after DBI and peaked at 8 h with the relative expression （9.365 2±0.543 8） 2^-Δ△Ct,then decreased gradually.S100A10 began to decrease at 4 h after DBI,and its expression level was the lowest around 8 h,with the expression level of （0.446 9±0.007 8） 2^-Δ△Ct,followed by an upward trend.The C3d mRNA expression of mice in the groups 4 h,8 h,12 h,1 d,3 d,5 d and 7 d after DBI was higher than that in the normalcontrol group,and the S100A10 mRNA expression of mice in the groups 4 h,8 h,3 d,5 d and 7 d after DBI was lower than that in the normal control group,with statistical significance （P<0.05）.Conclusions Within 7 days after DBI,A1s and A2s showed unimodal expression,and the trend was opposite,which is expected to become a biological index for estimation of injury time and treatment of DBI,and provide a theoretical basis for clinical medication and treatment.更多还原