摘要
目的探讨补肾活血方对兔髓核间充质干细胞(nucleus pulposus-derived mesenchymal stem cells,NPMSCs)移植治疗退变椎间盘P 38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,P 38 MAPK)信号通路的影响。方法选取128只SPF级健康新西兰大白兔,其中2例用于NPMSCs的获取、纯化及培养,选取24只为假手术组(A组);兔退变椎间盘模型成功100只,模型组(B组,24只)、补肾活血方组(C组,24只)、NPMSCs移植组(D组,24只)、补肾活血方+NPMSCs移植组(E组,24只),死亡6只。每组在治疗前、治疗后1周、治疗后2周、治疗后3周4个时间点均处死6只兔,取相应椎间盘,采用酶联免疫测定每组大兔不同时间点椎间盘P 38 MAPK、基质金属蛋白酶3(matrix metalloproteinase 3,MMP-3)、基质金属蛋白酶7(matrix metalloproteinase 7,MMP-7)的表达。结果(1)各组变化趋势:A组、B组组内不同时间点P 38 MAPK、MMP-3、MMP-7蛋白相对表达差异无统计学意义(P>0.05);C组、D组、E组P 38 MAPK、MMP-3、MMP-7蛋白相对表达与治疗前、治疗后1周、治疗后2周、治疗后3周呈下降趋势,各组组内差异有统计学意义(P<0.01)。(2)与A组比较:B组、C组、D组、E组P38MAPK、MMP-3、MMP-7蛋白相对表达在治疗前、治疗后1周、治疗后2周、治疗后3周均升高,组间差异有统计学意义(P<0.01)。(3)与B组比较:C组、D组、E组P 38 MAPK、MMP-3、MMP-7蛋白相对表达在治疗前差异无统计学意义(P>0.05);在治疗后1周、治疗后2周、治疗后3周均降低,组间差异有统计学意义(P<0.01)。(4)与C组比较:D组P 38 MAPK、MMP-3、MMP-7蛋白相对表达在治疗前、治疗后1周、治疗后2周、治疗后3周时同期差异无统计学意义(P>0.05);E组P 38 MAPK、MMP-3、MMP-7蛋白相对表达在治疗后1周、治疗后2周、治疗后3周均降低,组间差异有统计学意义(P<0.01)。(5)与D组比较:E组P 38 MAPK、MMP-3、MMP-7蛋白相对表达在治疗后1周、治疗后2周、治疗后3周均降低,组间差异有统计学意义(P<0.01)。结论补肾活血方对兔NPMSCs移植治疗退变椎间盘具有协同作用;其机制可能与抑制P 38 MAPK信号通路,下调MMP-3、MMP-7蛋白相关。
Objective To explore the effects of Bushen Huoxue formula on P 38 mitogen-activated protein(P 38 MAPK)kinase signal pathway in the treatment of degeneration intervertebral disc by the transplantation of nucleus pulposus-derived mesenchymal stem cells(NPMSC s).Methods 128 SPF grade healthy New Zealand white rabbits were selected,2 of which were used for the acquisition,purification and culture of NPMSCs,and 24 rabbits were selected as the sham operation group(group A).100 cases of rabbit model with degeneration intervertebral disc was successful,24 in model group(group B),24 in Bushen Huoxue formula group(group C),24 in NPMSCs transplantation group(group D),24 in Bushen Huoxue formula combined with NPMSCs transplantation group(group E),and 6 died.6 rabbits were sacrificed in each group at 4 time points,before,1,2 and 3 weeks after treatment.The corresponding intervertebral discs were taken.The expressions of P 38 MAPK,matrix metalloproteinase 3(MMP-3)and matrix metalloproteinase 7(MMP-7)in intervertebral disc at different time points in each group were determined by enzyme-linked immunoassay.Results(1)Change trend of each group:there was no statistically significant difference in the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 at different time points in group A and group B(P>0.05);the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group C,group D and group E showed a decreasing trend before,1,2 and 3 weeks after treatment,and the differences within each group were statistically significant(P<0.01).(2)Compared with group A,the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group B,group C,group D and group E were all higher than that in group A before,1,2 and 3 weeks after treatment,with statistically significant differences(P<0.01).(3)Compared with group B,there was no statistically significant difference in the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group C,group D and group E before treatment(P>0.05).1,2 and 3 weeks after treatment were all lower than the corresponding period of group B,and the difference between groups was statistically significant(P<0.01).(4)Compared with group C,there was no statistically significant difference in the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group D before,1,2 and 3 weeks after treatment(P>0.05).The relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group E were all lower than those in group C at 1,2 and 3 weeks after treatment,with statistically significant differences(P<0.01).(5)Compared with group D,the relative protein expression levels of P 38 MAPK,MMP-3 and MMP-7 in group E were lower than those in group E at 1,2 and 3 weeks after treatment,with statistically significant differences(P<0.01).Conclusion Bushen Huoxue formula has synergistic effect on NPMSCs transplantation in the treatment of degeneration intervertebral disc,and its mechanism may be related to the inhibition of P 38 MAPK signal pathway and the down-regulation the protein expression levels of MMP-3 and MMP-7.
作者
刘奕
陈晓峰
刘逸
蔡东岭
LIU Yi;CHEN Xiaofeng;LIU Yi;CAI Dongling(Second Department of Orthopedics,Panyu Hospital of Chinese Medicine,Guangzhou 511400,China)
出处
《环球中医药》
CAS
2020年第6期958-963,共6页
Global Traditional Chinese Medicine
基金
广东省中医药局中医药科研项目(20191262)
广州市番禺区科技计划医疗卫生项目(2019-Z04-64)。
关键词
补肾活血方
髓核间充质干细胞移植
退变椎间盘
P
38丝裂原活化蛋白激酶信号通路
基质金属蛋白酶3
基质金属蛋白酶7
Bushen Huoxue formula
Nucleus pulposus-derived mesenchymal stem cells
Degeneration intervertebral disc
P 38 mitogen-activated protein kinase signaling pathway
Matrix metalloproteinase-3
Matrix metalloproteinase-7
作者简介
刘奕(1980-),本科,主治中医师。研究方向:脊柱外科临床研究。E-mail:66144370@qq.com。
