目的观察补阳还五汤(BYHWD)及其四类有效部位(生物碱、苷、多糖、苷元)对脊髓损伤(SCI)大鼠大脑运动皮质神经元的影响,探究其神经保护作用的可能机制。方法 SD雄性大鼠48只,随机分为8组:正常组、假手术组、SCI组、BYHWD组、生物碱组、苷组、多糖组、苷元组。通过在T3-T4之间横断右侧半脊髓制备SCI模型,分别于术前1 d及术后1 d、1 w、4 w、8 w运用BBB评分评定大鼠后肢运动功能;术后8 w用TUNEL法检测神经细胞凋亡。结果SCI组与药物干预各组BBB评分均明显低于同期正常组、假手术组(P〈0.01);术后4 w,BYHWD组和生物碱组大鼠的BBB评分高于SCI组(P〈0.05);术后8w,BYHWD组、生物碱组和苷组大鼠的BBB评分高于SCI组(P〈0.05)。术后8 w,SCI组及药物干预各组大鼠细胞凋亡率明显高于正常组和假手术组(P〈0.01)。BYHWD组、生物碱组和苷组大鼠的细胞凋亡率低于SCI组(P〈0.01或P〈0.05);生物碱组、苷组、多糖组、苷元组的细胞凋亡率高于BYHWD组(P〈0.01或P〈0.05)。结论 BYHWD具有改善SCI大鼠后肢运动作用,其有效部位中的生物碱、苷可能为起效的主要物质基础,该作用机制可能与拮抗SCI大鼠运动皮质神经元凋亡有关。
Objective To observe the effect of Buyang Huanwu decoction(BYHWD) and its four kinds of active fractions(alkaloid, glycoside, polysaccharide and aglycone) on cerebral motor cortex in spinal cord injury(SCI) rats, and then explore the proper mechanism of the neuroprotection. Methods A total of 48 adult Sprague-Dawley rats were randomly divided into eight groups: control group, sham-operation group, SCI group, BYHWD group, alkaloid group, glycoside group,polysaccharide group and aglycone group(n=6). SCI model was performed by cutting the right half spinal cord at the level of T3-T4. Motor function of lower limb was assessed with Basso-Beattie-Bresnahan(BBB) scale at the following moments respectively: 1 d before operation, and 1 d, 1 w, 4 w and 8 w after operation. Ratio of cell apoptosis in cerebral motor cortex was detected by TUNEL staining after 8w treatment. Results Compared with the control group and sham-operation group,the BBB scores in SCI and all medicine intervention groups decreased significantly(P〈0.01). After four weeks of operation,the BBB scores in BYHWD and alkaloid groups were significantly higher than the SCI group(P〈0.05). After eight weeks of operation, the BBB scores in BYHWD, alkaloids and glycoside groups were higher than that in SCI group(P 0.05). After eight weeks of operation, the ratio of cell apoptosis in SCI and all medicine intervention groups was significantly higher than the normal and sham-operation groups(P〈0.01). The ratio of cell apoptosis in BYHWD, alkaloid and glycoside groups was lower than that in SCI group(P 0.01 or P 0.05); while ratio of cell apoptosis in alkaloid, glycoside, polysaccharide and aglycone groups were significantly higher than that in BYHWD group(P〈0.01 or 0.05). Conclusion BYHWD can promote the recovery of the motor function in SCI rats. The alkaloids and glycoside may be the most important important material base of BYHWD. Its mechanism may be associated with inhibiting neuronic apoptosis of SCI rats