曲克芦丁脑蛋白水解物对大鼠脊髓损伤后神经功能保护与修复研究

doi:10.3877/cma.j.issn.2095-9141.2018.06.010

目的

观察曲克芦丁脑蛋白水解物（TCH）注射液对脊髓损伤大鼠神经功能的保护作用，探讨其神经保护作用的可能机制。

方法

180只Sprague-Dawley（SD）大鼠采用随机数字表法分为5组，每组36只。A组为假手术组，B、C、D、E为模型组，采用"标准脊髓损伤打击装置"建立T10脊髓损伤（spinal cord injury，SCI）大鼠模型。A组不做干预；B组为空白对照组，术后1 mL生理盐水静脉注射，1次/d；C、D、E组术后分别使用曲克芦丁注射液、脑蛋白水解物注射液、TCH注射液静脉注射，0.0009 mL/g，加生理盐水配置成1 mL注射液，1次/d，至观察终点。比较各组术后1、3、7、14 d SCI大鼠后肢运动功能；观察术后5、24 h及3 d的SCI大鼠脊髓组织病理变化；比较各组术前与术后1、3、7、14 d的乙酰胆碱酯酶（AChE）和酸性磷酸酶（ACP）水平。

结果

术后7 d与14 d，E组与D组后肢功能评分显著增高，高于B、C两组（均P<0.05)。术后14 d，脊髓组织苏木精-伊红（HE)染色显示，E组较其他各组炎性细胞浸润、水肿、坏死程度均轻微，神经细胞存活较多。术后C、E组伊文氏蓝（EB)含量和脊髓组织含水量显著低于B、D两组（均P<0.05）。术后3、7、14 d，D、E组AChE含量恢复显著，高于B组（均P<0.05）；术后7与14 d，E组显著高于D组（均P<0.05）。术后3、7与14 d，D、E组ACP含量均显著低于B组（均P<0.05）。

结论

TCH可改善SCI大鼠后肢运动功能，保护损伤的脊髓神经并促进脊髓神经功能恢复。

关键词: 脊髓损伤, 曲克芦丁脑蛋白水解物, 神经保护, 乙酰胆碱酯酶, 酸性磷酸酶

Objective

To investigate the protective effect of troxerutin and cerebroprotein hydrolysate (TCH) on neural function after spinal cord injury, and to explore its possible mechanism of protective effects.

Methods

"Standard Spinal Cord Injury Strike Device" was employed to establish SCI rat model. A total of 180 SD rats were randomly divided into group A (sham-operated group, n=36), group B (SCI+Normal saline, n=36), group C (SCI+Troxerutin group, n=36), group D (SCI+Cerebroprotein hydrolysate, n=36), group E (SCI+TCH, n=36). Each group was treated with the corresponding intervention, 0.0009 mL/g, 1 time per day after the operation by caudal vein injection, respectively. The behavior function was evaluated. The histological injure of spinal cord was assessed. The expression of AChE and ACP were detected too.

Results

On the 7^th and 14^th day after operation, the BBB score in D and E groups was higher than that in the other groups significantly (P<0.05). On the 14^th day after operation, the infiltration, edema of inflammatory cells, and the degree of necrosis in group E were much lighter, and the survival number of nerve cells was more than that in other groups. The spinal cord edema and blood spinal cord barrier in C and E groups were improved better significantly than those in the other groups. On the 3^rd, 7^th and 14^th day, AChE content in D and E groups raised significantly more than that in group B (all P<0.05); The ACP content in group D and group E was significantly lower than that in group B (P<0.05).

Conclusion

TCH could play a role of protecting the neural function of SCI rat and contribute to the recovery of injurious spinal cord nerve.

Key words: Spinal cord injury, Troxerutin and cerebroprotein hydrolysate, Neuroprotection, Acetylcholinesterase, Acid phosphatase

表1 脊髓损伤后不同时点各组大鼠BBB运动功能评分比较（n=5，±s，分）

组别	术前	术后1 d	术后3 d	术后7 d	术后14 d
A组	21.00	21.00±0.00	21.00±0.00	21.00±0.00	21.00±0.00
B组	21.00	1.20±0.45^a	1.80±0.45^a	3.60±1.14^a	5.80±1.48
C组	21.00	1.20±1.10^a	2.40±1.67^a	4.60±1.14^a	8.00±0.71^ab
D组	21.00	1.00±0.00^a	3.80±0.84^ab	7.40±0.89^abc	11.00±1.58^abc
E组	21.00	1.20±0.45^a	3.00±0.71^ab	7.80±0.84^abc	10.00±1.58^abc
F值	-	1231.438	399.762	299.463	111.130
P值	-	0.001	0.001	0.001	0.001

表1 脊髓损伤后不同时点各组大鼠BBB运动功能评分比较（n=5，±s，分）

组别	术前	术后1 d	术后3 d	术后7 d	术后14 d
A组	21.00	21.00±0.00	21.00±0.00	21.00±0.00	21.00±0.00
B组	21.00	1.20±0.45^a	1.80±0.45^a	3.60±1.14^a	5.80±1.48
C组	21.00	1.20±1.10^a	2.40±1.67^a	4.60±1.14^a	8.00±0.71^ab
D组	21.00	1.00±0.00^a	3.80±0.84^ab	7.40±0.89^abc	11.00±1.58^abc
E组	21.00	1.20±0.45^a	3.00±0.71^ab	7.80±0.84^abc	10.00±1.58^abc
F值	-	1231.438	399.762	299.463	111.130
P值	-	0.001	0.001	0.001	0.001

图1 脊髓HE染色（HE×200）

图2 血液脊髓屏障与脊髓水肿状况观察

图3 脊髓组织AchE与ACP表达水平

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Protective effect of troxerutin and cerebroprotein hydrolysate on neural function in spinal cord injury rat

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Protective effect of troxerutin and cerebroprotein hydrolysate on neural function in spinal cord injury rat