复方三七消痛软膏超声导入对兔膝骨性关节炎治疗作用及机制研究

Study of Therapeutic Effects and Mechanisms of Ultrasound Phoresis Compound Sanqi Xiaotong Paste for Treatment Knee Osteoarthritis in Rabbits

目的:探讨复方三七消痛软膏用超声波(US)导入对兔KOA疗效及其作用机制。方法:将4月龄新西兰大白兔40只按随机数字表法分为复方三七消痛软膏超声波导入组(A组)、普通US组(B组)、舒筋止痛水外擦组(C组)、假US组(D组)和正常对照组(E组)各8只,除E组外各组动物均行双侧前交叉韧带离断术造模。6周后A、B、C、D组分别给予复方三七消痛软膏超声波导入普通US、假US、舒筋止痛水外擦干预10 d,E组不给予任何干预。干预后软骨切片HE染色光镜观察形态结构及Mankin评分;采用TUNEL法检测软骨细胞凋亡阳性比率;采用TUNEL法检测软骨细胞凋亡阳性率,Western-blot技术检测软骨细胞caspase-9、caspase-3蛋白水平表达。结果:A组关节软骨病变明显轻于B、C、D组但较E组严重;A组Mankin评分低于B、C、D组、高于E组,差异有统计学意义。干预后A组软骨细胞凋亡阳性率以及caspase-9和caspase-3蛋白表达均低于B、C、D组,高于E组,差异有统计学意义。结论:复方三七消痛软膏超声波导入能更好地延缓KOA软骨退变,其机制可能是通过抑制caspase-9和caspase-3表达从而抑制软骨细胞凋亡实现。

Objective：Observing the theraputic effects and studying the mechanisms of US phoresis Compound Sanqi Xiaotong Paste in KOA rabbits. Methods：Four month old, 40 white New Zealand rabbits, were randomly divided into US phoresis Compound Sanqi Xiaotong Paste （A group）, normal US （B group）, ShuJinZhiTong water local external pat （C group ） , sham normal US （D group） and nornal controls （E group ） groups. All were subjected to double knee anterior cruciate ligament abscise （ACLA） except E group, each group included 8 rabbits. Six weeks after surgery, A, B, C, D group （surgical knee joint） were interrupted with US phoresis Compound Sanqi Xiaotong Paste, normal US, ShuJinZhiTong water local external pat, sham normal US for 8 min per day for 10 days （5 days after exposure, stopping 2 days, continue, exposure 5 days） , respectively. After 10 days intervention, all rabbits were sacrificed by air injection. The cartilaginous tissue was stained with Hematoxylin and Eosin （H＆E） and observed chondrocyte morphology by light microscopy and Mankin＇ s score were counted. Choudrocytes apoptosis were detected by TUNEL. Incide tibial plate full-thickness cartilage for apotposis gene of caspase-9 and caspase-3, measurement the protein content by Western-Blot in chondrocyte. Results The Mankin scores in A groups was significantly lower than B, C, D groups, and significantly higher than E group. The apoptosis of chondrocytes and the caspase-9, caspase-3 protien expression in A groups were significantly lower than B, C, D groups, and significantly higher than E group. Conclusions US phoresis Compound Sanqi Xiaotong Paste can regress cartilage apomorphosis for treatment OA, and superior to the general US interruption. It downregulated the expression of caspase-9 and caspase-3 protien in chondrocyte, which it may be the theraputic signal molecules.