红景天苷调节细胞自噬对脑出血后继发性损伤的作用及其机制研究

Treatment of secondary injury after cerebral hemorrhage is crucial for functional recovery of central nerve system. Apoptosis is involved in the secondary injury after cerebral hemorrhage. Whether salidroside, which is effective in treating cerebral hemorrhage, can affect autophagy has not been reported. We will study the role of autophagy in the secondary injury after cerebral hemorrhage in the animal model induced by autologous blood injection in the brain. The rats will be divided into 6 groups: normal group, sham operated control group，model group，rapamycin plus salidroside group, 3-MA plus salidroside group and salidroside group.We will evaluate the number of autophagic vascule and autolysosome at the different time point after intracerebral injection of autologous blood by electric microscopy, immunohistochemistry and confocal laser microscope. Expression of LC3I, LC3II, Beclin1,Cathepsin D,P62,LAMP-1 and LAMP-2 will be detected by immunohistochemistry. Based on these data we will elucidate the development of authphage after cerebral hemarrhage and the effect of salidroside on autophage in treating cerebral hemorrhage, which will provide new evidence for treating cerebral hemorrhagic patients with salidroside.

防治继发性损伤是治疗脑出血的关键，细胞凋亡参与了脑出血的继发性损伤，而与细胞凋亡密切相关的自噬在脑出血发生后所起的作用及能否被红景天苷所调节，未见报道。红景天苷具有良好的神经保护作用，但作用机制尚不明了。本课题通过脑内注射自体血建立大鼠脑出血模型，设立正常对照组、模型对照组、假手术组、红景天苷干预组、红景天苷+雷帕霉素干预组及红景天苷+3-甲基腺嘌呤霉素干预组，采用电镜、蛋白免疫印迹、免疫双标激光共聚焦显微镜检测不同时间点神经细胞增殖和凋亡、自噬体和自噬溶酶体、自噬相关蛋白LC3、LC3II/LC3I的比值及Beclin 1、Cathepsin D、P62、LAMP-1和LAMP-2的表达，了解脑出血后自噬的发生和发展规律及其在脑出血过程中所起的作用，阐明红景天苷调节细胞自噬对脑出血继发性损伤的作用及其机制，为红景天苷治疗脑出血提供科学的理论基础和实验依据。

防治继发性损伤是治疗脑出血的关键，细胞凋亡参与了脑出血的继发性损伤，而与细胞凋亡密切相关的自噬在脑出血发生后所起的作用及能否被红景天苷所调节，未见报道。红景天苷具有良好的神经保护作用，但作用机制尚不明了。本课题采用电镜、光镜、蛋白免疫印迹、免疫组化检测神经细胞增殖和凋亡、自噬体和自噬溶酶体、自噬相关蛋白LC3、LC3II/LC3I的比值及Beclin 1、Cathepsin D、P62的表达，了解脑出血后自噬的发生和发展规律及其在脑出血过程中所起的作用。研究结果显示：脑出血后血肿周围自噬被激活，自噬泡及自噬相关蛋白表达增多，红景天苷通过调节自噬从而对脑出血后细胞周围发挥神经保护作用，阐明了红景天苷调节细胞自噬对脑出血继发性损伤的作用及其机制，为红景天苷治疗脑出血提供科学的理论基础和实验依据。