维格列汀对大鼠脊髓损伤后神经细胞凋亡及运动功能恢复的影响

The effect of vildagliptin on neural cell apotosis and locomotor recovery in a rat model of spinal cord injury

目的:探究维格列汀(Vilda)对大鼠脊髓损伤(SCI)后神经细胞凋亡及运动功能恢复的影响。方法:雌性SD大鼠随机分成3组(n=16):Sham组、SCI组、SCI+Vilda组;采用血管夹压迫法建立大鼠SCI模型,成模后SCI+Vilda组按10mg·kg^-1·d^-1的剂量灌胃给药,SCI组大鼠给予等量0.9%氯化钠溶液;在术后第1、第3、第7、第14、第21、第28天进行后肢运动功能恢复情况评估(BBB评分);术后第3天提取脊髓组织进行组织免疫荧光染色检测Cleaved Caspase 3,提取组织蛋白进行Western blot检测Bcl-2、Bax、Cleaved Caspase3蛋白的表达情况;术后第7天提取脊髓组织进行尼氏染色检测存活神经元;术后第28天,对各组大鼠进行足印迹步态分析。结果:与Sham组比较,SCI组BBB评分显著降低,步态不一致,脊髓前角运动神经元数量减少,免疫荧光显示Cleaved Caspase3表达增强,Western blot显示Bax、Cleaved Caspase 3表达上升,Bcl-2表达下降(P<0.05);与SCI组比较,SCI+Vilda组BBB评分明显提升,步态一致性更好,脊髓前角运动神经元数量增加,Cleaved Caspase 3荧光表达减弱,Bax、Cleaved Caspase 3表达下降,Bcl-2表达上升(P<0.05)。结论:Vilda可抑制SCI大鼠脊髓神经细胞的凋亡,从而促进大鼠SCI后运动功能的恢复。

Objective: To investigate the effect of Vildagliptin (Vilda) on locomotor function recovery and neural cell apotosis in a rat model of spinal cord injury. Methods: Forty-eight SD rats were randomly divided into 3 groups (n=16): the Sham group, the SCI group and the SCI+Vilad group. Clip compression SCI model was established as previously described. The SCI+Vilad group was treated with Vilad at a dose of 10 mg·kg^-1·d^-1 by gavage for 7 days in a row, then once every 3 days;the SCI group was treated with the same dosage of normal saline. The Basso-Beattie-Bresnahan (BBB) score was obtained at 1, 3, 7, 14, 21, 28 d after operation. Immunofluorescence staining and Western blot were used to detect the expression of apoptotic related proteins expression (Bcl-2, Bax, Cleaved Caspase 3) at 3 d after operation. Nissl staining was performed at 7 d after operation to detect the surviving neurons. At 28 d, the, the footprint analysis was used to assess the locomotor recovery. Results: Compared with the Sham group, SCI group showed significantly lower BBB scores and inconsistent footprints;the number of VMNs was decreased significantly. Immunofluorescence staining and western blot results showed enhanced expression of apoptotic related proteins (Bax, Cleaved Caspase 3), but the expression of anti-apoptotic related protein (Bcl-2) was decreased (P<0.05). Compared with the SCI group, the rats in SCI+Vilda group got higher BBB scores, more consistent footprints;the number of VMNs was also increased. At the same time, the expression of apoptotic related proteins (Bax, Cleaved Caspase 3) was decreased significantly and the expression of anti-apoptotic related protein (Bcl-2) was increased obviously (P<0.05). Conclusion: All of these findings indicated that Vildagliptin shows neuroprotective effect after SCI in rats via inhibiting the VMNs apoptosis, and finally promotes the locomotor recovery.