3个道地产区丹参混合提取物对大鼠脑缺血再灌注后神经细胞凋亡的影响  

Effecs of Danshen mixed extract from 3 genuine producing areas on nerve cells apoptosis in cerebral ischemia and reperfusion rats

作　　者：李红刚[1] 王红鸽[1] 聂晓枫[1] 孙 陈卫银[2] 

机构地区：[1]陕西省渭南职业技术学院医学院临床教研室,陕西渭南714000 [2]成都中医药大学附属医院神经内科,四川成都610072

出　　处：《河北中医》2016年第10期1515-1519,共5页Hebei Journal of Traditional Chinese Medicine

基　　金：渭南职业技术学院院级科研项目(编号:WZYY201322)

摘　　要：目的通过观察山东、河南、四川3个道地产区丹参混合提取物对大鼠局灶性脑缺血再灌注后脑组织B细胞淋巴瘤-2基因(Bc1-2)、肿瘤抑制基因p53的影响,探讨3个道地产区丹参对神经细胞凋亡的影响。方法将60只SD大鼠随机分为正常组、假手术组、模型组、山东丹参组、河南丹参组、四川丹参组,每组10只。用线栓法阻断大脑中动脉,建立大鼠局灶性脑缺血再灌注损伤模型,用苏木精-伊红(HE)染色法观察缺血脑组织的病理学变化,采用免疫组化法检测再灌注24 h后各组Bc1-2和p53蛋白的表达。结果正常组和假手术组细胞与间质之间界限清楚,细胞核完整,核仁清晰;模型组细胞与间质之间界限不清楚,大部分组织细胞坏死,多数细胞溶解呈空泡状,可见核固缩和核碎裂;3个道地产区丹参组脑组织细胞坏死程度均较模型组轻,可见少量细胞核固缩及溶解,部分胞质液化。正常组、假手术组Bcl-2的积分光密度值(IOD)较少,二者比较差异无统计学意义(P>0.05);模型组Bcl-2的IOD值较正常组、假手术组增加(P<0.05);与模型组比较,3个道地产区丹参组均能上调Bcl-2的IOD值(P<0.05);与河南、四川丹参组比较,山东丹参组上调Bcl-2的IOD值的幅度最大(P<0.05)。正常组、假手术组p53的IOD值较少,二者比较差异无统计学意义(P>0.05);模型组p53的IOD值较正常组、假手术组增加(P<0.05);与模型组比较,3个道地产区丹参组均能降低p53的IOD值(P<0.01);与河南丹参组比较,山东、四川丹参组降低p53的IOD值的幅度最大(P<0.05)。结论 3个道地产区丹参混合提取物均能通过上调Bcl-2表达、降低p53表达,从而抑制神经细胞凋亡,发挥脑保护作用。不同产区丹参混合提取物对各指标表达影响不同,其原因可能与不同产区丹参混合提取物所含活性成分的含量不同,作用靶点不一致有关。Objective To observe the effecs of Danshen mixed extract from 3 genuine producing areas（Shandong,Henan,Sichuan） on B-cell lymphoma-2 gene（Bcl-2） and tumor suppressor gene p53 in cerebral ischemia and reperfusion rats,to investigate the effects Danshen of three difference area on nerve cells apoptosis.Methods 60 SD rats were randomly divided into normal group,shame operation group,model group,Shandong Danshen group,Henan Danshen group and Sichuan danshen group,10 rats in each group. The rats model of focal cerebral ischemia-reperfusion were established with the middle cerebral artery occlusion. The pathologic change of cerebral ischemia tissue was observed by HE staining. The expressions of Bc1-2 and p53 after 24 reperfusion were observed by mean of immunohistochemistry. Results Cells in normal and shame operation group had clear margin with mesenchyme, integral nucleus and clear nucleolus.Cells in model group had unclear margin with mesenchyme,many cytolysis showed vacuole-shape,and the karyopyknosis and nuclear fragmentation were observed. The necrotic degree of brain tissue cells in three genuine producing areas were lighter than model group,there were few karyopyknosis and karyolysis,partial plasmatosis. The integrated optical density（IOD） of Bcl-2 in normal and shame operation group was fewer,with no statistical difference between two groups（P〉0.05）. The IOD of Bcl-2 in model group was increased as compared with the normal and shame operation group（P〈0.05）. As compared with model group,the IOD of Bcl-2 in three genuine producing areas were up-regulation（P〈0.05）. As compared with Henan and Sichuan danshen group,the IOD of Bcl-2 in Shandong Danshen group had maximum enhanced amplitude（P〈0.05）. The IOD of p53 in normal and shame operation group was fewer,with no statistical difference between two groups（P〉0.05）. The IOD of p53 in model group was increased as compared with the normal and shame operation group（P〈0.05）. As compared with model group,the IOD of p53 in t

关 键 词：疾病模型 动物 脑缺血 神经元 细胞学 细胞凋亡 再灌注损伤 丹参 

分 类 号：R-332[医药卫生] R322