丹酚酸B联合间充质干细胞移植修复退变椎间盘的研究

Treatment of degenerative intervertebral disc with salvianolic acid B combined with MSCs transplantation

使用针吸髓核法建立新西兰大白兔的椎间盘退变模型,随机分为8组,分别是A^E组。A组为单纯骨髓间充质干细胞(MSCs)移植组;B组为MSCs-丹酚酸B联合治疗组,包括B1、B2、B3、B4分别为1.25、2.50、5.00、10.00mg·L-14个亚组;C组为磷酸盐缓冲液(PBS)移植组;D组为假移植组;E组为正常对照组。8周时处死动物,分别采用半定量免疫组织化学法和间苯三酚法测定各组Ⅱ型胶原含量和椎间盘蛋白多糖含量,探讨丹酚酸B联合MSCs移植修复退变椎间盘的治疗效果。结果表明:B3、B4组Ⅱ型胶原灰度值低于A组,Ⅱ型胶原含量和蛋白多糖含量均高于A组,差异显著。说明丹酚酸B浓度大于等于5.00mg·L^(-1)时,丹酚酸B联合干细胞移植的效果优于单纯干细胞移植组。

The intervertebral disc degeneration model were established by the aspiration of nucleus pulposus in the New Zealand white rabbits. The successful models were divided into 8 groups randomly. Group A was treated with transplan- tation of the bone marrow mesenchymal stem cells （MSCs）. Group B was treated by MSCs-salvianolic acid B. Group 13 contained four subgroups such asB1, 1.25 mg· L-1, B2, 2,50 mg· L-1, B3, 5.00 mg· L-1, B4, 10.00 mg· L-1. The four subgroups were treated by the MSCs suspension with the above content of salvianolic acid B respectively. Group C was treated with PBS transplantation. Group D was treated with sham graft. Group E was normal control group. At the end of 8th week, the rabbits were sacrificed and the content of type II collagen in each group was determined by semi quantitative immunohistochemistry, and the content of proteoglycan in each group was measured by the Benzene-three- phenol method to investigate the efficacy of salvianolic acid B combined with MSCs transplantation in the repair of degen- erative intervertebral disc. Immunohistochemical gray values of type II collagen among subgroups B3, B4 were lower than that of Group A. The content of type II collagen and proteoglycan among subgroups B3, B4 were higher than that of group A. The difference was statistically significant. Salvianolie acid B （≥5.00 mg· L-1） plus MSCs transplantation contributed to repair degenerative intervertebral disc, and they were more effective than MSCs transplantation alone.