目的:观察老龄大鼠局灶性脑缺血后DNA损伤修复相关基因GADD45表达的规律,探讨海风藤酮对神经细胞凋亡及GADD45表达的影响。方法:采用26月龄Wistar大鼠,经光化学诱导局灶性脑缺血后应用海风藤酮进行干预,观察不同时间和空间状态下GADD45蛋白表达的改变.同时运用TUNEL染色观察神经细胞的凋亡现象。结果:缺血4h组,在缺血中心区可见少许GADD45反应阳性细胞,而缺血周边区GADD45反应阳性细胞大量表达,较对照组差异有统计学意义(P〈0.05)。缺血24h组,周边区GADD45表达较缺血4h有所降低,但仍高于假手术组(P〈0.05)。至缺血5d时,周边区GADD45表达较假手术组无明显差异。海风藤酮治疗组。缺血周边区TUNEL阳性细胞数较假手术组增多(P〈0.05)。但较对照组减少(P〈0.05),而GADD45表达阳性细胞数较对照组增多(P〈0.05),与假手术组比较增多伊〈0.011。结论:海风藤酮可有效保护缺血神经细胞,同时可以上调DNA修复基因GADD45的表达,减小DNA损伤的程度。
Objective: To observe the expression of DNA damage and repair gene: GADD45 following cerebral ischemia in aged rats, and to investigate the effects of Kadsurenone on the expression of GADD45 and neuronal cell apoptosis. Methods: Wistar rats of 26-month-old with focal cerebral ischemia were administered herb Kadsurenone. The change of GADD45 protein expression was determined with immunohistochemistry, and the neuronal cell apoptosis with TUNEL staining techniques. Results: Few GADD45 response positive cells existed in the core of ischemic area 4 hours after ischemia, but a great deal expressed in the peripheral area, which was significantly different from the control group (P〈0.05). GADD45 response positive cells at 24 hours were fewer than those at 4 hours, but higher than those in pseudo-surgery group(P〈 0.05). TUNEL positive cells in herb Kadsurenone group increased compared with pseudo-surgery group (P〈 0.05), but decreased compared with 24-hour ischemia control group (P〈0.05). GADD45 positive cell of herb kadsurenone group increased compared with control group (P〈0.05) and pseudo-surgery group (P〈0.01). Conclusions: Kadsurenone, an inhibitor of PAF has protective effect on cerebral ischemia, and it can decrease the extent of DNA damage and up-regulate the expression of GADD45, a DNA repair gene.