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目的探讨胶质细胞生长因子2(GGF2)对大鼠液压打击颅脑损伤的神经保护作用。方法采用侧方液压打击装置制备大鼠颅脑损伤模型。将 pEGFP-N1-GGF2表达质粒或 pEGFP-N1载体质粒与阳离子脂质体混合后,脑内直接注射转染大鼠脑组织。将34大鼠随机分为4组,分别为治疗组(pEGFP-N1-GGF2+脂质体,n=10)、载体对照组(pEGFP-N1载体+脂质体,n=10)、脂质体对照组(脂质体,n=10)和假手术组(n=4)。伤后连续观察爬坡、平衡和行走试验,于伤后第10天处死大鼠取脑组织,进行 HE、尼氏及 MBP、NSE、GFAP 免疫组化染色。结果伤后第5d,治疗组大鼠行为学指标的恢复优于载体对照组和脂质体对照组[爬坡试验(角度):66.25±3.54 vs 58.31±3.72、57.21±3.93,P<0.05;平衡试验(评分):2.59±0.21 vs 3.41±0.25、3.24±0.22,P<0.05;行走试验(s):20.15±2.59 vs 27.00±3.47、27.80±3.00,P<0.05],以行走实验改善最为明显。伤后第10天,治疗组大鼠的皮层、海马神经元计数多于载体对照组和脂质体对照组(皮层外颗粒和外锥体层:98±10 vs 75±7、67±8,P<0.05;皮层内锥体层:37±4 vs 19±3、23±4,P<0.05;海马 CA1区:102±11 vs67±8、58±9,P<0.01),皮层、皮层下白质 MBP 染色信号强于对照组。结论阳离子脂质体介导GGF2基因治疗能有效地促进大鼠颅脑损伤后的恢复。
Objective To investigate the neuroprotective effect of glial cell growth factor 2 (GGF2) on brain injury induced by fluid percussion in rats. Methods A rat model of craniocerebral injury was prepared by lateral hydraulic strikes. The pEGFP-N1-GGF2 expression plasmid or pEGFP-N1 vector plasmid was mixed with cationic liposomes and then injected into the brain of rats by direct injection into the brain. The 34 rats were randomly divided into 4 groups: treatment group (pEGFP-N1-GGF2 + liposome, n = 10), vehicle control group (pEGFP-N1 vector + liposome, n = 10), liposome Control (liposome, n = 10) and sham-operated (n = 4) groups. The rats were sacrificed on the 10th day after injury, and the brain tissues were harvested for HE, Nissl, MBP, NSE and GFAP immunohistochemical staining. Results On the 5th day after injury, the recovery of behavioral indexes in the treatment group was superior to that of vehicle control group and liposome control group [hill climbing test (angle): 66.25 ± 3.54 vs 58.31 ± 3.72, 57.21 ± 3.93, P <0.05; Balancing test (score): 2.59 ± 0.21 vs 3.41 ± 0.25, 3.24 ± 0.22, P <0.05; walking test: 20.15 ± 2.59 vs 27.00 ± 3.47, 27.80 ± 3.00, P <0.05] obvious. On the 10th day after injury, the counts of cortical and hippocampal neurons in the treated group were more than those in the vehicle control group and the liposome control group (extracranial granules and outer pyramidal layer: 98 ± 10 vs 75 ± 7, 67 ± 8, P <0.05; cortical pyram: 37 ± 4 vs 19 ± 3, 23 ± 4, P <0.05; hippocampal CA1 region: 102 ± 11 vs 67 ± 8, 58 ± 9, P <0.01) White matter MBP staining signal stronger than the control group. Conclusion Cationic liposome-mediated GGF2 gene therapy can effectively promote the recovery after traumatic brain injury in rats.