胶原酶溶解椎间盘髓核的体外实验  

In vitro experiment of collagenase dissolving nucleus gelatinosus

作　　者：陈延娥[1] 郭广洲[2] 王毅平[3] 康春阳[3] 

机构地区：[1]北京大学深圳医院检验科,广东省深圳市518034 [2]深圳市第五人民医院中心实验室 [3]深圳市第五人民医院,广东省深圳市518001

出　　处：《中国临床康复》2006年第25期138-139,157,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:通过胶原酶对椎间盘髓核的体外溶解实验,进一步验证胶原酶溶盘术的作用并确认胶原酶的用量。方法:实验于2005-03/10在深圳市第五人民医院中心实验室完成,实验采用无菌操作的方法,对临床收集的北京大学深圳医院和深圳市第五人民医院的10例腰椎间盘突出患者手术取出的髓核组织(均为自愿捐献),使用临床治疗用胶原酶,以胶原酶浓度为2.5,5,10,15mkat/L分为4组,在37℃的条件下,对髓核组织进行体外溶解试验,分别在培养3,6,9,15,30d收集上清液,采用碱水解法检测溶解液中羟脯氨酸含量。结果:①在相同时间条件下,胶原酶浓度为2.5mkat/L组分解髓核组织产生羟脯氨酸的量与胶原酶浓度为5mkat/L组、胶原酶浓度为10mkat/L组、胶原酶浓度为15mkat/L组间存在差异(培养3d:5.53,11.47,13.17,10.09μg/g;培养6d:6.93,13.48,13.68,10.09μg/g;培养9d:8.0,15.29,15.44,11.48μg/g;培养15d:9.56,19.33,17.12,12.77μg/g;培养30d:12.08,24.29,28.28,22.59μg/g,P<0.05),而5mkat/L组、胶原酶浓度为10mkat/L组分解髓核组织产生羟脯氨酸的量与胶原酶浓度为15mkat/L组之间没有差异(P>0.05)。②当胶原酶浓度>5mkat/L时,其分解胶原纤维产生羟脯氨酸的量并没有随着其浓度的增加而增加。结论:羟脯氨酸的变化较好的反映胶原酶对髓核的溶解作用,胶原酶对椎间盘突出髓核的溶解过程是一个渐进缓慢的过程,在一定的范围外(>5mkat/L)的胶原酶作用下,其产生羟脯氨酸的量没有随胶原酶浓度的增加而增加。AIM： To further validate the effect of collagenasse chemonucleolysis and the amount of collagenase through in vitro experiment of collagenase dissolving nucleus gelatinosus. METHODS： This experiment was carried out at the Central Laboratory of the Fifth People＇s Hospital of Shenzhen City from March. to October 2005. Nucleus pulpesus tissue voluntarily donated by 10 patients with lumbar intervertehral disc herniation from Shenzhen Hospital of Beijing University and the Fifth People＇s Hospital of Shenzhen City were collected with aseptic operation method. Collagenaso was used in the clinical therapy. Collagenase was divided into 4 groups of 2.5,5,10,15 mkat/L collagenase, separately. At 37℃, in vitro solubility test of nucleus pulposus tissue was conducted, and the supematant was collected on days 3,6,9,15,30 after culture, then, the content of hydroxyproline in the solution was detected with alkaline hydrolysis method. RESULTS： （1） During the same time period, there was difference of amount of hydroxyproline produced in decomposition of nucleus pulposus tissue between 2.5 mkat/L collagenase group and 5 kat/L collagenase group or 10 mkat/L collagenase group or 15 mkat/L collagenasc group （On day 3 after culture：5.53,11.47,13.17,10.09 μg/g; On day 6 after culture： 6.93, 13.48, 13.68, 10.09 μg/g; On day 9 after culture： 8.0, 15.29, 15.44, 11.48μg/g; On day 15 after culture： 9.56, 19.33, 17.12, 12.77 μg/g; On day 30 after culture：12.08,24.29,28.28, 22.59 μg/g, P 〈 0.05）,while there was no difference of the amount of hydroxyproline produced in decomposition of nucleus pulposus tissue between 5 mkat/L collagenase group or 12 mkat/L collagenase group and 15 mkat/L collagenasc group（P 〉 0.05）. （2） When the content of collagenasc was 〉 5 mkat/L, the amount of hydroxyproline produced in decomposition of collagenous fibers was not increased with the increase of concentration. CONCLUSION： The change of hydroxyproline can well reflect the dissolution of collagenase for nucleus

关 键 词：胶原酶类 羟脯氨酸 基质金属蛋白酶 

分 类 号：R681[医药卫生—骨科学]