针刀松解颈周腧穴对大鼠退变颈椎间盘细胞外基质金属蛋白酶-1、3及其抑制因子-1基因表达的影响与髓核超微结构观察

Effect of Acupotomy Lysis at Cervical Acupoints on Expression of Matrix Metalloproteinases and Tissue Inhibitor of Metalloproteinase 1 and Changes of Pulpiform Nucleus Ultrastructure in Rats with Degenerated Cervical Intervertebral Discs

目的:通过观察针刀松解颈周腧穴对大鼠退变颈椎间盘细胞外基质金属蛋白酶-1、3(MMP-1、MMP-3)及其抑制因子-1(TIMP-1)基因表达的影响及髓核超微结构,探讨其缓解颈椎间盘退变的作用机制。方法:参照动静力失衡性大鼠颈椎间盘退变造模方法,将造模成功大鼠随机分为模型组、颈周腧穴组、夹脊穴组、药物组,并设空白对照组,每组15只。夹脊穴组取C2-7椎旁夹脊穴,颈周腧穴组取颈周的双"脑空"穴、"脑户"穴、"大椎"穴、双"曲垣"穴、双"天宗"穴,进行针刀干预,每5d治疗1次,各治疗3次;药物组采用布洛芬胶囊+颈复康颗粒灌胃,每天1次,治疗10d。各组继续喂养20d后处死采集标本,用定量多聚酶链反应检测大鼠颈椎间盘MMP-1、MMP-3及TIMP-1的基因表达,并在电镜下行髓核超微结构观察。结果:MMP-1、MMP-3的基因表达检测,模型组高于空白组(P<0.05),颈周腧穴组、夹脊穴组、药物组低于模型组(P<0.05),颈周腧穴组、夹脊穴组低于药物组(P<0.05);TIMP-1的基因表达检测,模型组低于空白组(P<0.05),颈周腧穴组、夹脊穴组高于模型组及药物组(P<0.05)。髓核超微结构观察:椎间盘髓核退变程度为空白组<颈周腧穴组/夹脊穴组<药物组<模型组。结论:针刀松解颈周腧穴可通过调控MMP-1、MMP-3及TIMP-1的基因表达,影响椎间盘细胞外基质的合成和降解,从而延缓椎间盘退变。

Objective To observe the effect of acupotomy lysis at acupoints around the neck on expression of matrix metalloproteinase 1（MMP-1）,MMP-2and tissue inhibitor of metalloproteinase 1（TIMP-1）genes and ultrastructure of pulpiform nucleus in cervical intervertebral disc degeneration（IVDD）rats,so as to explore its mechanism underlying easing IVDD.Methods SD rats were randomly allocated to control（n=15）,model（n=14）,Jiaji（EX-B 2,n=13）,cervico-acupoint（n=14）and medication groups（n=14）.The cervical IVDD model was established by using static-dynamic imbalance method.For rats of the Jiaji（EX-B 2）and cervico-acupoint groups,EX-B 2-points of the cervical 2-7segments,and peri-cervical acupoints：bilateral＂Naokong＂（GB 19）,＂Naohu＂（GV 17）,＂Dazhui＂（GV 14）,bilateral＂Quyuan＂（SI 13）and bilateral＂Tianzong＂（SI 11）were separately punctured with a needle-knife,once every 5days for 3times,and for rats of the medication group,Brufen Capsules（15mg·kg-1·d-1）and Jingfukang Granule（0.5mg·kg-1·d-1）were given by intragastric administration,once daily for 10 days.The expression levels of MMP-1,MMP-3and TIMP-1genes in the pulpiform nucleus of cervical intervertebral discs were detected by RT-PCR and changes of the ultrastructure of the pulpiform nucleus observed under transmission electron microscope.Results Compared to the control group,the expression levels of MMP-1mRNA and MMP-3mRNA of the cervical intervertebral disc tissues were significantly up-regulated in the model group（P〈0.05）,and that of TIMP-1mRNA was obviously down-regulated in the model group（P〈0.05）.After the treatment,the increased expression of MMP-1mRNA and MMP-3mRNA and the decreased expression of TIMP-1mRNA were reversed by acupotomy lysis and medication（P〈0.05）except TIMP-1mRNA in the medication group（P〉0.05）.No significant differences were found between the Jiaji（EX-B 2）and cervicoacupoint groups in down-regulating MMP-1 mRNA and MMP-3 mRNA expression and up-regulating TIMP-1 mRNA expression（P〉0.05）.Results of electron microscope examinations showed that the ultrastructural injury changes of cells of the pulpiform nucleus were relatively milder in the Jiaji（EX-B 2）and cervico-acupoint groups,followed by the medication group in comparison with those of the model group.Conclusion Acupotomy lysis at acupoints around the neck can improve the ultrastructural changes of cells of the pulpiform nucleus of cervical intervertebral discs in IVDD rats,which is possibly by regulating the expression of MMP-1,MMP-3and TIMP-1genes.