益气化瘀补肾方对D-半乳糖诱导小鼠腰椎间盘衰老的影响

Effect of "Yiqi Huayu Bushen Decoction" on Senescence of Lumbar Intervertebral Disc Induced by D-galactose in Rats

目的:观察益气化瘀补肾方对D-半乳糖诱导小鼠腰椎间盘衰老的影响。方法:8周龄雄性健康C57BL/6J小鼠30只,随机分为空白对照组、诱导盐水灌胃组及益气化瘀补肾方组。空白对照组皮下注射生理盐水,其余2组均皮下注射D-半乳糖复制腰椎间盘衰老模型;盐水灌胃组灌胃等容量生理盐水,益气化瘀补肾方组灌胃中药水溶液。连续给药10周后处死小鼠,取下腰部椎间盘,进行免疫组化分析以及实时定量RT-PCR检测。结果:免疫组化染色显示,益气化瘀补肾方可以明显降低软骨终板及纤维环中p16INK4a和p27KIP蛋白表达;实时定量RT-PCR分析发现,益气化瘀补肾方可显著增加Bmi1、CDK4和C-myc mRNA的表达,降低p16INK4a、cyclin D1、RB、p19ARF、Mdm2和p53 mRNA表达。结论:益气化瘀补肾方具有延缓D-半乳糖诱导小鼠腰椎间盘衰老的作用,并且这种延缓作用与调控衰老相关基因及细胞周期相关蛋白表达相关。

Objective： To observe the effect of ＂Yiqi Huayu Bushen Decoction＂ on senescence of lumbar intervertebral disc induced by D-galactose in rats.Methods： Thirty male rats were randomized into control group,saline group and Chinese medicine group.Rats in control group were subcutaneously injected by saline,and the rats in other two groups were subcutaneously injected by D-galactose everyday.Meanwhile,rats in Chinese medicine group were intragastrically administrated with ＂Yiqi Huayu Bushen Decoction＂,and saline group administrated with saline.The rats were sacrificed after administration for 10 weeks continuously,and the lumbar intervertebral disc samples were used to do the immunohistochemistry analysis and RT-PCR detection.Results： The immunohistochemical staining showed that ＂Yiqi Huayu Bushen Decoction＂ could decrease the cartilage endplate and expression of p16INK4a and p27KIP in fibrous ring;RT-PCR analysis showed that ＂Yiqi Huayu Bushen Decoction＂ could increase the expression of Bmi1,CDK4 and C-myc mRNA,and decrease the expression of p16INK4a,cyclin D1,RB、p19ARF,Mdm2 and p53mRNA.Conclusion： ＂Yiqi Huayu Bushen Decoction＂ can delay the senescence of lumbar intervertebral disc,which relates to relative genes and protein expression of relative cells.