补肾活血汤干预衰老大鼠退变椎间盘模型中经典WNT信号通路β-catenin蛋白含量的研究

Effects of Bushen Huoxue Decoction on β-catenin Protein Level of WNT Signal Pathway in Lumbar Intervertebral Disc Degeneration Aging Model Rats

目的观察补肾活血汤对衰老大鼠退变椎间盘组织形态结构及β-catenin蛋白含量的影响。方法选取三月龄健康SD大鼠80只,雌雄各半,随机分为空白组、假模型组、模型组、中药低剂量组以及中药高剂量组,每组16只,雌雄各半。除空白组和假模型组外,其余各组注射D-半乳糖建立大鼠衰老动物模型,空白对照组、假模型组与模型组每日生理盐水灌胃,中药低剂量组及中药高剂量组补肾活血汤每日灌胃,持续12周。分别于造模后、灌胃4周后、灌胃8周后、灌胃12周后每组随机抽取雌雄大鼠各2只处死取材,光镜下观察椎间盘组织形态学变化并运用蛋白质印迹法(Western Blot)检测β-catenin蛋白表达差异。结果与模型组比较,经补肾活血汤干预后,中药低剂量组及中药高剂量组大鼠椎间盘组织形态学评分降低(P<0.05),β-catenin蛋白表达水平降低(P<0.05)。结论补肾活血汤能有效抑制经典Wnt/β-catenin信号通路中关键蛋白分子β-catenin蛋白的表达,从而起到延缓椎间盘退变的作用。

Objective To observe the effect of Bushen Huoxue decoction on the structural and histopathological changes andβ-catenin protein in lumbar intervertebral disc degeneration aging model rats.Methods Eighty three-month old SD rats(male-female half and half)were randomly divided into blank control group,sham-model group,model group,TCM low-dose group and TCM high-dose group,16 rats in each group.Except the blank group and sham-model group,the aging model rats in other groups were established by injecting D-galactose.The rats in blank control group,sham-model group and model group were daily given normal saline by gavage.TCM low-dose and high-dose groups were daily treated with Bushen Huoxue decoction for 12 weeks by gavage.Two male rats and two female rats in each group were randomly executed at after modelling,after 4 weeks of gavage,after 8 weeks of gavage and after 12 weeks of gavage,and samples were selected from dead rats.The histomorphological change of model group was observed by light microscope,and the expression ofβ-catenin protein was detected by Western Blot method.Results Compared with the model group,the histomorphology score in TCM low-dose and high-dose groups treated with Bushen Huoxue decoction decreased(P<0.05),and the expression ofβ-catenin protein decreased(P<0.05).Conclusion Buyang Huanwu decoction shows inhibition effect on the expression of keyβ-catenin protein in the classical Wnt/β-catenin signal pathway,and thus could relieve the degeneration of intervertebral disc.