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目的:通过检测实验性自身免疫性脑脊髓炎(EAE)模型脊髓中MMP-9、ICAM-1、HO-1的mRNA表达来研究苦参素对EAE的治疗作用。方法:选取40只大鼠随机分为EAE组、空白对照组、苦参素低剂量组和苦参素高剂量组。EAE组和苦参素两个剂量组注射豚鼠全脊髓匀浆(GPSCH)抗原乳剂免疫建立EAE模型。苦参素两个剂量组于免疫当天起鼠尾静脉注射苦参素注射液200 mg·kg-1和100 mg·kg-1。观察3组大鼠的发病情况,采用5分评分法对症状进行评分。设计和合成三条针对MMP-9、ICAM-1、HO-1mRNA的特异性引物,采用PCR技术检测苦参素治疗16 d后大鼠脊髓中MMP-9、ICAM-1、HO-1mRNA的表达。结果:EAE组平均症状评分(2.45±0.36)高于苦参素治疗组(1.35±0.21),且两者之间存在显著性差异(P<0.05)。与空白对照组比较,EAE组大鼠脊髓中MMP-9、ICAM-1的表达显著升高(P<0.01),而苦参素治疗组中MMP-9、ICAM-1的表达与EAE组比较有所降低(P<0.05)。与空白对照组比较,EAE组大鼠脊髓中HO-1的表达显著降低(P<0.05),而苦参素治疗组中HO-1的表达显著升高(P<0.01)。结论:苦参素能够通过降低MMP-9和ICAM-1mRNA的表达,并促进HO-1mRNA的表达而发挥对实验性自身免疫性脑脊髓炎的治疗作用。
OBJECTIVE: To investigate the therapeutic effect of oxymatrine on EAE by detecting the mRNA expression of MMP-9, ICAM-1 and HO-1 in the spinal cord of experimental autoimmune encephalomyelitis (EAE) model. Methods: Forty rats were randomly divided into EAE group, blank control group, low dose oxymatrine group and high dose oxymatrine group. EAE group and oxymatrine two dose groups injected guinea pig whole spinal cord homogenate (GPSCH) antigen emulsion immune EAE model. Oxymatrine two dose groups from the tail of the mouse tail vein injection of oxymatrine injection of 200 mg · kg-1 and 100 mg · kg-1. The incidence of the rats in the three groups was observed, and the score was scored by a 5-point scale. Three specific primers targeting MMP-9, ICAM-1 and HO-1 mRNA were designed and synthesized. The expression of MMP-9, ICAM-1 and HO-1 mRNA in the rat spinal cord were detected by PCR after 16 days of oxymatrine treatment. Results: The mean symptom scores of EAE group (2.45 ± 0.36) were significantly higher than that of oxymatrine treatment group (1.35 ± 0.21), with significant difference (P <0.05). Compared with the blank control group, the expression of MMP-9 and ICAM-1 in the EAE group was significantly increased (P <0.01), while the expression of MMP-9 and ICAM-1 in the Oxymatrine treatment group was significantly higher than that in the EAE group Decreased (P <0.05). Compared with the blank control group, the expression of HO-1 in the spinal cord of EAE group was significantly decreased (P <0.05), while the expression of HO-1 in the oxymatrine treatment group was significantly increased (P <0.01). Conclusion: Oxymatrine can exert the therapeutic effect on experimental autoimmune encephalomyelitis by decreasing the expression of MMP-9 and ICAM-1 mRNA and promoting the expression of HO-1 mRNA.