丹红注射液对蛛网膜下腔出血后脑血管痉挛防治作用的实验研究

发布时间：2018-12-20 15:51

【摘要】：目的：观察大鼠蛛网膜下腔出血（Subarachnoid hemorrhage,SAH）模型的脑血管形态学变化及血管壁蛋白激酶C（Protein kinase C,PKC）的表达情况，探讨丹红注射液对蛛网膜下腔出血后脑血管痉挛(Cerebralvasospasm,CVS)的防治作用及机制。方法：将56只SD大鼠随机分为4组：正常对照组（n=8），生理盐水组（n=16），SAH组（n=16）及SAH后丹红注射液治疗组（治疗组，n=16）。正常对照组不予任何处理，7天后处死；生理盐水组以枕大池注入生理盐水后分别于3、7天处死（各8只）；SAH组及SAH后丹红注射液治疗组均采用自体股动脉血枕大池二次注射法建模，建模后SAH组亦于3、7天处死（各8只）；治疗组于每日腹腔注射丹红注射液后分别于3、7天处死（各8只）。建模后分别按各时间点取出动物基底动脉脑桥段，切片后观察血管壁的厚度及血管壁的形态变化，采用免疫组化法测定血管壁PKC的表达情况，结果进行统计学分析。结果： 1.HE染色：正常对照组和生理盐水组基底动脉管壁平滑,内皮细胞结构完整,内弹力板未见皱褶或断裂。SAH组术后第3、7天均有基底动脉管腔缩小，血管壁增厚且结构紊乱；内膜出现皱褶或断裂,厚薄不均，内皮细胞变形、肿胀；中、外膜变厚，平滑肌细胞肥大，出现炎性细胞浸润，，以第7天时更严重。治疗组各时间点基底动脉管腔缩小、血管壁增厚、内膜皱褶及炎性细胞浸润等病理变化均较SAH组有所减轻。基底动脉内径与管壁厚度：正常组与生理盐水组比较基底动脉内径与管壁厚度无明显变化（P0.05）；SAH组和治疗组的基底动脉内径较正常组均有缩小,管壁厚度较正常组均有增加。SAH组与治疗组均出现基底动脉内径缩小、管壁厚度增加，SAH组第7天比第3天更明显，而治疗组第3天比第7天稍明显。在各时间点，SAH组与生理盐水组比较，基底动脉内径及管壁厚度有显著差异（P0.05）；治疗组与SAH组比较，基底动脉内径较大，管壁较薄，差异有统计学意义（P0.05）。 2.免疫组化：在正常组和生理盐水组探测到很弱的PKC免疫阳性反应；在SAH组和治疗组均可见PKC的免疫阳性反应可以在血管内膜，平滑肌层内观察到，呈棕黄或棕褐色，定位于细胞胞浆内和胞膜上。PKC的免疫阳性反应在SAH组第7天时最为显著，第3天时次之；正常组和生理盐水组PKC表达比较无统计学意义(P0.05)，治疗组PKC表达各时间点较SAH组明显减弱，且均有统计学意义（P0.05）。结论： 1.蛛网膜下腔出血后脑血管痉挛时基底动脉PKC的活性表达有明显的升高，其程度与脑血管痉挛形态学改变的程度一致，表明PKC的活性增加在脑血管持续痉挛中有着重要作用。 2.丹红注射液可能是通过抑制PKC的活性来缓解蛛网膜下腔出血后的脑血管痉挛。
[Abstract]:Objective: to observe the changes of cerebral vascular morphology and the expression of vascular wall protein kinase (C (Protein kinase) in rats with subarachnoid hemorrhage (Subarachnoid hemorrhage,SAH). To investigate the preventive effect and mechanism of Danhong injection on cerebral vasospasm (Cerebralvasospasm,CVS) after subarachnoid hemorrhage. Methods: Fifty-six SD rats were randomly divided into 4 groups: normal control group (n = 8), normal saline group (n = 16) and danghong injection group (n = 16). The normal control group was killed after 7 days without any treatment, and in the saline group, 8 rats in each group were killed on the 7th day after injection of normal saline into the cistern of the occipital cistern. The SAH group and the SAH hindanghong injection treatment group were treated with the second injection of autologous femoral arterial blood occipital cistern, and the SAH group was killed on the 7th day after modeling (8 rats each). In the treatment group, danghong injection was injected intraperitoneally every day. After modeling, the animal basilar artery brain bridge segment was taken out according to each time point, the thickness of vascular wall and the morphological changes of vascular wall were observed after slicing, and the expression of PKC in vascular wall was determined by immunohistochemical method. The results were statistically analyzed. Results: 1.HE staining: the basilar artery wall was smooth, the endothelial cell structure was intact, and the inner elastic plate was not creased or broken in the normal control group and normal saline group. The basilar artery lumen was reduced in the SAH group on the 7th day after operation. The wall of blood vessel was thickened and the structure was disordered. The intima was creased or broken, uneven in thickness, deformed and swollen in endothelial cells, thickening of the outer membrane, hypertrophy of smooth muscle cells, inflammatory cell infiltration, and more serious on the 7th day. The pathological changes of basilar artery lumen, wall thickening, intimal crease and inflammatory cell infiltration at each time point in the treatment group were reduced compared with those in the SAH group. The diameter and thickness of basilar artery: there was no significant change in diameter and wall thickness of basilar artery between normal group and normal saline group (P0.05). The diameter of basilar artery in SAH group and treatment group was smaller than that in normal group, and the thickness of basilar artery was increased in SAH group and treatment group, and the thickness of basilar artery was increased in SAH group and treatment group. The thickness of basilar artery in SAH group was more obvious on the 7th day than on the third day. In the treatment group, the 3rd day was a little more obvious than the 7th day. At each time point, SAH group and normal saline group, basilar artery diameter and wall thickness were significantly different (P0.05); treatment group and SAH group, basilar artery diameter was larger, wall thin, the difference was statistically significant (P0.05). 2. Immunohistochemistry: weak PKC immunoreactivity was detected in normal group and normal saline group. The immunoreactivity of PKC was observed in the intima and smooth muscle layer of the blood vessels in both SAH and treatment groups. It was located in the cytoplasm and on the membrane of the cells. The immunoreactivity of PKC was the most significant in the SAH group on the 7th day. The third day was the second; There was no significant difference in PKC expression between normal group and normal saline group (P0.05). The expression of PKC in treatment group was significantly lower than that in SAH group (P0.05). Conclusion: 1. The expression of PKC activity in basilar artery was significantly increased during cerebral vasospasm after subarachnoid hemorrhage, which was consistent with the morphological changes of cerebral vasospasm. It indicated that the increase of PKC activity played an important role in cerebral vasospasm. 2. Danhong injection may relieve cerebral vasospasm after subarachnoid hemorrhage by inhibiting the activity of PKC.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.35

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