人参皂苷Rg1对大鼠局灶性脑缺血再灌注损伤后脑皮层内泛素修饰蛋白聚集的影响

doi:10.13481/j.1671-587x.20190201

摘要/Abstract

摘要： 目的：探讨人参皂苷Rg1在大鼠脑缺血再灌注（I/R）损伤后皮层内泛素修饰蛋白聚集过程中的作用，进一步阐明人参皂苷Rg1对脑I/R损伤的治疗机制。方法：将大鼠采用线栓法栓塞1.5 h制作大脑中动脉缺血（MCAO）模型。72只健康雄性SD大鼠分为假手术组，I/R模型组，阳性药物对照（尼莫地平）组，低、中和高剂量（10、20和40 mg·kg^-1）人参皂苷Rg1组，每组12只大鼠，均采用腹腔注射给药。在建模24h后采用TTC染色法和Longa法测量各组大鼠脑梗死面积和神经功能缺损评分，HE染色观察各组大鼠脑缺血后皮层和海马内的神经元死亡情况，免疫组织化学和Western blotting法检测各组大鼠皮层内泛素修饰蛋白聚集物的表达情况。结果：与I/R模型组比较，尼莫地平和剂量人参皂苷Rg1组大鼠脑梗死面积百分比明显减小（P<0.05），神经功能缺损评分明显降低（P<0.05）。HE染色，与假手术组比较，I/R模型组大鼠神经元稀疏，出现碎裂、溶解现象；与I/R模型组比较，尼莫地平组和各剂量人参皂苷Rg1大鼠神经元核碎裂、溶解、粉染等现象均有不同程度改善。免疫组织化学检测，与假手术组比较，I/R模型组大鼠泛素修饰蛋白表达水平明显增多（P<0.05）；与I/R模型组比较，尼莫地平和各剂量人参皂苷Rg1组大鼠泛素修饰蛋白阳性表达水平均明显降低（P<0.05），以高剂量人参皂苷Rg1组效果最明显。Western blotting法检测，与假手术组比较，I/R模型组大鼠泛素修饰蛋白聚集物水平明显升高（P<0.05）；与I/R模型组比较，尼莫地平和各剂量人参皂苷Rg1组大鼠泛素修饰蛋白聚集物水平明显降低（P<0.05），且以高剂量人参皂苷Rg1组效果最为明显。结论：人参皂苷Rg1可抑制大鼠脑皮层内I/R损伤所诱导的泛素修饰蛋白聚集物形成，进而减轻大鼠脑I/R损伤。

关键词: 脑缺血再灌注, 人参皂苷, 蛋白聚集, 尼莫地平

Abstract: Objective: To explore the effect of ginsenoside Rg1 on the ubiquitin-modified protein aggregation in the cortex after cerebral ischemia reperfusion(I/R) injury in the rats, and to further clarify the therapeutic mechanism of ginsenoside Rg1 in the cerebral I/R injury.Methods: The middle cerebral artery occlusion (MCAO) model was set up with suture method for 1.5 h of embolization. A total of 72 rats were divided into sham operation group, I/R model group,positive drug control (nimodipine) group, low, middle,and high doses(10, 20, and 40 mg·kg^-1) of ginsenoside Rg1 groups. All 12 rats in each group were given intraperitoneal injection. TTC staining and Longa's score method were used to detect the infarction areas and the neurological deficit scores of the rats in various groups 24 h after modeling. The death of neurons in the cortex and hippocampus after cerebral ischemia of the rats in various groups were observed with HE staining.Immunohistochemistry and Western blotting method were used to detect the expression of ubiquitin-modified protein aggregation in the cortex of the rats in various groups.Results: Compared with I/R group, the percentages of infarction areas of the rats in nimodipine group and ginsenoside Rg1 groups were significantly decreased(P<0.05). and the neurological deficit scores were decreased(P<0.05).The HE staining results showed that compared with sham operation group, the neurons in I/R model group were sparse, showing fragmentation and dissolution;compared with I/R model group, the phenomena of cell nucleus fragmentation, dissolution and powder staining in nimodipine group and different doses of ginsenoside Rg1 groups were all improved to different degrees. The immunohistochemical results showed that compared with sham operation group, the positive expression level of ubiquitin-modified protein in I/R model group was increased significantly(P<0.05); compared with I/R model group,the positive expression levels of ubiquitin-modified protein in nimodipine group and different doses of ginsenoside Rg1 groups were decreased (P<0.05),especially in high dose of ginsenoside Rg1 group (P<0.05). The Western blotting results showed that compared with sham operation group, the level of ubiquitin-modified protein aggregates in I/R model group was significantly increased(P<0.05);compared with I/R model group, the levels of ubiquitin-modified protein aggregates in nimodipine group and different doses of ginsenoside Rg1 were decreased(P<0.05), especially in high dose of ginsenoside Rg1 group.Conclusion: Ginsenoside Rg1 can inhibit the formation of ubiquitin-modified protein aggregates induced by I/R injury in the cortex, thereby alleviating the I/R injury in the rats.

Key words: brain ischemia-reperfusion, ginsenosides, protein aggregation, nimodipine

中图分类号:

R473.74

郑天洋, 许海洋, 金日华, 姜宏. 人参皂苷Rg1对大鼠局灶性脑缺血再灌注损伤后脑皮层内泛素修饰蛋白聚集的影响[J]. 吉林大学学报(医学版), 2019, 45(02): 217-222.

ZHENG Tianyang, XU Haiyang, JIN Rihua, JIANG Hong. Effect of ginsenoside Rg1 on cortical ubiquitin-modified protein aggregation after cerebral ischemia-reperfusion in rats[J]. Journal of Jilin University(Medicine Edition), 2019, 45(02): 217-222.

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