补肾活血方对兔退变椎间盘模型MMP-3表达影响

Effect of Bushen Huoxue Decoction on MMP-3 Expression in Rabbit Defunct Intervertebral Disc Model

目的观察补肾活血方对兔退变椎间盘组织形态结构及MMP-3表达的影响。方法选取3月龄健康兔共40只,随机分为5组,每组8只,依次为对照组、模型组、中药低剂量组、中剂量组和高剂量组,除正常对照组外,其余兔均行椎间盘退变造模术。造模后3 d开始给药,对照组及模型组每日以生理盐水灌胃代替中药,中药低、中、高剂量组补肾活血方每日灌胃,连续给药8周。采用HE染色、免疫组化染色等检测方法观察椎间盘组织形态学变化,并运用PCR法检测MMP-3表达水平。结果HE染色显示:药物干预后,低剂量组髓核细胞数量变化不明显,纤维环及髓核存在裂隙,中、高剂量组细胞数量较低剂量组显著增加;免疫组化染色显示:与模型组相比,中药低剂量组MMP-3表达未见明显变化(P>0.05);中剂量组和高剂量组MMP-3的表达均有显著下调(P<0.05);PCR法检测法显示:与模型组相比,中药干预后,椎间盘组织内增高的MMP-3表现出降低趋势,其降低程度与药物剂量呈正比,中剂量组及高剂量组的MMP-3表达量有统计学差异(P<0.05)。结论造模成功后MMP-3表达量显著上调,经补肾活血方干预后可抑制MMP-3的表达,说明补肾活血方可在一定程度上起到延缓椎间盘退变的作用。

Objective To observe the effect of Bushen Huoxue Decoction on the morphological structure and MMP-3 expression of defunct intervertebral disc in rabbits.Methods A total of 40 healthy rabbits aged 3 months were selected and randomly divided into 5 groups,8 rabbits in each group,respectively,the control group,the model group,the low-dose group of traditional Chinese medicine,the medium-dose group and the high-dose group.In addition to the normal control group,all the other rabbits underwent disc degeneration surgery.Three days after the model was established,the control group and the model group were given normal saline daily instead of traditional Chinese medicine,and the low-dose,medium-dose and high-dose groups were given Bushen Huoxue Decoction daily for 8 weeks.The morphological changes of the intervertebral disc were observed by HE staining and immunohistochemical staining,and the expression level of MMP-3 was detected by PCR.Results HE staining showed that,after drug intervention,the number of nucleus pulposus cells in the low-dose group did not change significantly,and there were cracks in the annulus fibrosus and nucleus pulposus.Immunohistochemical staining showed no significant change in MMP-3 expression in the low-dose group compared with the model group(P>0.05).The expression of MMP-3 in the medium dose group and the high dose group was significantly decreased(P<0.05).PCR method showed that compared with the model group,MMP-3 increased in the intervertebral disc tissues showed a decreasing trend after TCM intervention,and the degree of decrease was positively proportional to the drug dose.There was a statistical difference in MMP-3 expression between the medium dose group and the high dose group(P<0.05).Conclusion The expression of MMP-3 was significantly up-regulated after the successful modeling,and the expression of MMP-3 could be inhibited after the prognosis of Bushen Huoxue Decoction,suggesting that invigorating kidney and activating blood circulation could delay the degeneration of intervertebral disc to a certain extent.