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目的:观察补阳还五汤(buyanghuanwu decoction,BYHWD)对脊髓损伤(spinal cord injury,SCI)模型大鼠脊髓神经细胞凋亡相关蛋白Bcl-2、Bax及其mRNA表达的影响,初步探讨BYHWD抑制脊髓神经细胞凋亡的作用机制。方法:Wistar大鼠40只,随机分为四组:正常对照组、SCI模型组、BYHWD组和甲基强的松龙(MP)注射组,每组10只大鼠;后三组动物先建立SCI损伤模型,然后分别给予BYHWD组和MP组BYHWD和MP治疗。四组动物分别于术后1、7、14、21 d和28 d对大鼠后肢神经功能的恢复情况进行BBB(Basso-Beattie-Bresnahan)功能评分。第28 d处死各组大鼠,采用Real-time PCR法检测脊髓组织Bcl-2和Bax mRNA的表达情况,Western Blot法检测脊髓组织Bcl-2和Bax蛋白的表达情况。结果:大鼠麻醉清醒后,SCI组大鼠损伤平面以下完全瘫痪。BBB评分结果显示:术后第7 d~28 d,与正常对照组相比,SCI组BBB评分明显降低(P<0.05);与SCI组相比,BYHWD处理组和MP处理组大鼠BBB评分明显增高(P<0.05),但BYHWD处理组和MP处理组大鼠相比,BBB评分无明显差异(P>0.05)。Real-time PCR和Western Blot结果显示:与正常对照组相比,SCI组Bax蛋白及mRNA的表达明显升高(P<0.05),而Bcl-2蛋白及mRNA的表达则明显有所降低(P<0.05)。与SCI组相比,BYHWD处理组和MP处理组大鼠脊髓组织Bax蛋白及mRNA表达明显降低(P<0.05),而Bcl-2蛋白及mRNA表达则明显有所升高(P<0.05)。但BYHWD处理组和MP处理组大鼠脊髓组织Bcl-2、Bax蛋白及mRNA表达之间无明显差异(P>0.05)。结论:BYHWD可以改善SCI大鼠后肢的运动功能,通过下调Bax的表达,上调Bcl-2的表达,从而抑制SCI大鼠脊髓神经细胞的凋亡,发挥其保护作用。
OBJECTIVE: To observe the effect of buyanghuanwu decoction (BYHWD) on the expression of Bcl-2, Bax and its mRNA in spinal cord nerve cells of spinal cord injury (SCI) model rats, and to investigate the effect of BYHWD inhibition Mechanism of apoptosis of spinal cord nerve cells. Methods: Forty Wistar rats were randomly divided into four groups: normal control group, SCI model group, BYHWD group and methylprednisolone (MP) injection group, 10 rats in each group. The last three groups of animals were established SCI injury model, and then were given BYHWD group and MP group BYHWD and MP treatment. The BBB (Basso-Beattie-Bresnahan) score was used to evaluate the recovery of neurological function in the hindlimbs at 1, 7, 14, 21 d and 28 d after surgery. The rats in each group were sacrificed on the 28th day. The expression of Bcl-2 and Bax mRNA in the spinal cord was detected by Real-time PCR. The expression of Bcl-2 and Bax protein in the spinal cord was detected by Western Blot. Results: After anesthetized rats awake, rats in SCI group were completely paralyzed below the level of injury. Compared with SCI group, BBB score of BYHWD group and MP group was significantly higher than that of SCI group (P <0.05). BBB score showed that BBB score of SCI group was significantly lower than that of control group from the 7th to 28th day after operation (P <0.05). However, there was no significant difference in BBB score between BYHWD group and MP group (P> 0.05). The results of Real-time PCR and Western Blot showed that compared with normal control group, the expression of Bax protein and mRNA in SCI group was significantly increased (P <0.05), while the expression of Bcl-2 protein and mRNA was significantly decreased (P <0.05). Compared with SCI group, Bax protein and mRNA expression in spinal cord of BYHWD group and MP treated group were significantly decreased (P <0.05), while Bcl-2 protein and mRNA expression was significantly increased (P <0.05). However, there was no significant difference in the expression of Bcl-2, Bax protein and mRNA between the BYHWD treatment group and the MP treatment group (P> 0.05). CONCLUSION: BYHWD can improve the motor function of hindlimb in SCI rats, and down-regulate the expression of Bax and up-regulate the expression of Bcl-2, thereby inhibiting the apoptosis of spinal cord nerve cells of SCI rats and exerting their protective effect.