精制蛇毒酶对大鼠缺血后肢比目鱼肌的影响及作用机制

目的研究精制蛇毒酶对大鼠不同时段缺血后肢比目鱼肌的影响及机制。方法成年Wistar大鼠150只,显微外科技术制作大鼠后肢缺血模型,随机分为空白对照组、缺血组及精制蛇毒酶治疗组。空白组仅切开皮肤后缝合;缺血组从术后1d开始按1.2ml/kg体重于大腿肌肉分四点等量注射生理盐水,每周3次,共计2w;治疗组于大腿肌肉分四点等量注射精制蛇毒酶(1mg/ml,1.2ml/kg体重)每周3次,治疗2w;分别于术后第3d、7d、14d、21d和28d采取缺血后肢及对侧的比目鱼肌。测定各组肌肉组织含水量、线粒体游离钙浓度和线粒体通透性转运通道开放程度(用吸光度变化ΔS表示)的动态变化。结果比目鱼肌含水程度在缺血1w内升高,随后下降,精制蛇毒酶治疗组比目鱼肌含水程度在造模术21、28d后显著高于缺血组(P<0.05)。在术后第14、21d,精制蛇毒酶治疗组线粒体游离钙浓度显著低于缺血组(P<0.05);精制蛇毒酶治疗组MPTP开放程度ΔS值在第14、21、28d均显著高于缺血组(P<0.05)。结论精制蛇毒酶能维持大鼠缺血后肢比目鱼肌含水量,降低缺血腓肠肌内线粒体游离钙浓度,抑制MPTP的开放程度。

The effect of pure snake venom on soleus in the ischemic hindlimb of rats

Objective To investigate the effect of pure snake venom on skeletal muscles in the ischemic hindlimb of rats.Methods 150 adult Wistar rats were divided into control group,ischemic group and pure snake venom treating group randomly.The rats were sacrificed at 3,7,14,21and 28 days after microsurgery technique.The soleus in the ischemia hindlimbs and the opposite normal hindlimbs were harvested,the water content,free calcium concentration of the mitochondrial,the opening level of mitochondria permeability transition pore(MPTP) were detected respectively.Results The water content of the soleus in the pure snake venom treating group was significantly higher than that in ischemic group at 21,28 days after the ischemic model made(<0.05).The free calcium concentration of the mitochondrial was significantly lower in treating group at 14,21 days after surgery.The opening level of MPTP(expressed by ΔS) of the gastroenemius in the pure snake venom treating group was significantly higher than that in the ischemic group at 14,21,28 days after surgery(<0.05).Conclusion Pure snake venom could maintain water content,decrease the concentration of free calcium in the mitochondrial and inhibit the opening level of MPTP in the ischemic soleus of rats.