青娥方对骨质疏松骨关节炎模型鼠早期软骨下骨基质金属蛋白酶3和骨桥蛋白表达的影响

Effect of Qing’e Decoction on the Expression of MMP3 and OPN in Subchondral Bone of Osteoarthritis Model Rats

目的:探讨中药复方青娥方对骨质疏松骨关节炎模型鼠早期软骨下骨组织形态以及基质金属蛋白酶3和骨桥蛋白表达的影响。方法:30只大鼠随机分为正常组(A组)、青娥方组(B组)、骨质疏松模型(C组)、骨质疏松骨关节炎模型(D组)和骨关节炎模型(E组),每组6只。其中B,C和D组进行骨质疏松模型制备,1个月后B,D和E组进行骨关节炎造模,接着术后第2天,B组灌服青娥方药液,其余各组灌服等量的生理盐水,1次/d,连续4周。结果:病理切片可见D组的各个时期均可见软骨下骨板逐渐增厚,软骨细胞四层结构不易分辨,可见双重潮线。软骨下骨板逐渐断裂至钙化软骨层内,软骨下骨骨小梁逐步紊乱、稀疏、断裂。B组干预后第1周软骨下骨板部分碎裂,潮标线不清晰,干预后第2周潮标线部分显示,可见少量增生血管。干预后第4周可观察到潮线结构基本完整,软骨下骨板下方的骨小梁结构欠规则,内可见增生的血管。软骨下骨骨组织在中药复方干预后第4周时各组软骨下骨骨小梁体积分数、骨小梁数量、骨小梁分离度和骨小梁厚度(Tb.Th)与A组差异无统计学意义(P>0.05),与D组差异有统计学意义(P<0.05)。胫骨髁骨组织中正常组MMP3和OPN基因维持在相对稳定的水平;B组MMP3和OPN基因上调表达维持在相对稳定的水平;与D组相比,B组MMP3和OPN在治疗后第2周和第4周差异有统计学意义(P<0.05)。结论:中药青娥方可以改变关节软骨和软骨下骨镜下表现,调节软骨下骨组织骨小梁各项指标以及MMP3和OPN表达水平,改善骨代谢,延缓早期软骨下骨骨丢失,防治骨质疏松骨性关节炎。

Objective:To investigate the effect of Qing’e decoction on the expression of MMP3 and OPN in the early subchondral bone of osteoarthritis model rats.Methods:30 rats were randomly divided into A(normal group),B(Qing’e decoction group),C(osteoporosis group),D(osteoporosis osteoarthritis group) and E(osteoarthritis group).The osteoporotic model was made in group B,C and D.One month later the osteoarthritis model was made in group B,D and E.Gavage administration was carried out with Qing’e decoction in group B.The other groups were given the same amount of saline with once a day for 4 weeks.Results:In group of different period,the subchondral bone plate became thickened gradually,and the four layers of chondrocytes were indistinguishable.The subchondral bone plate gradually ruptured into the calcified cartilage layer.The trabecula of subchondral bone was gradually disordered,sparse and broken.In the Qing’e decoction group,the subchondral bone plate was partly broken,and the tide mark was not clear after one week of intervention,and a small amount of hyperplastic vessels were revealed in the tide mark after two weeks’ of intervention.At the 4 th week after the intervention,the structure of tidal line was basically complete,the trabecular structure under the subchondral bone plate was irregular,and new proliferative vessels were fund in it.There was no significant difference in the volume fraction,the number,the separation degree and the thickness of trabecula(Tb.Th)at 4 weeks after the intervention of traditional Chinese medicine as compared with group A(P<0.05),and significant statistic difference was found between the B and D group(P>0.05).In the tibial condyle,the MMP3 and OPN genes in the normal group were maintained at a relatively stable level,the up-regulated expression of MMP3 and OPN genes in the Qing’e decoction group was maintained at a relatively stable level.As compared with the osteoporosis osteoarthritis group,the MMP3 and OPN in the Qing’e decoction group were significantly different at 2 and 4 weeks after treatment(P<0.05).Conclusion:Qing’e decoction can change the microscopic manifestations of articular cartilage and subchondral bone.It can regulate the indexes of bone trabecula,the expression level of MMP3 and OPN,improve bone metabolism,delay the early loss of subchondral bone and prevent osteoarthritis.