低频超声促透中药对兔膝关节炎细胞因子的影响

Influence of low-frequency ultrasound for enhancing permeation of Chinese medicinal on cytokines in rabbits with knee osteoarthritis

目的观察低频超声促透通络止痛汤对膝关节炎(KOA)动物模型兔血清、关节液中细胞因子的影响。方法将32只新西兰白兔随机分为正常组5只、模型组、超声促透中药组、超声促透盐水组各9只。将膝关节腔内注射1.6%木瓜蛋白酶溶液制备KOA的动物模型。经病理学切片证实造模成功后,后2组予干预治疗3周。在治疗结束后1周抽取血清、关节液,采用放射免疫的方法分别测定血清、关节液中细胞因子的含量;并处死所有动物,取滑膜、关节软骨,行HE染色显微镜下观察,并记录Mankin's组织学评分。结果 Mankin's评分显示试验组的滑膜组织评分与模型组比较,有显著性差异(P<0.05),而软骨细胞评分与模型组比较,无显著性差异(P>0.05)。对照组的滑膜组织、软骨细胞组织学评分与模型组比较,均无显著性差异(P>0.05)。与正常组相比,骨关节炎模型组血清、关节液内细胞因子的含量均显著升高(P<0.05);试验组关节液内4种炎性因子的含量与模型组相比,均呈显著减低(P<0.05),而血清中含量无统计学意义。结论低频超声促透通络止痛汤治疗实验性兔KOA的作用机制可能是通过减缓滑膜细胞的炎症反应,抑制关节液中细胞因子的生成,从而减轻膝关节局部炎性反应。

Objective To observe the influences of low-frequency ultrasound for enhancing permeation of Tongluo Zhitong Tang on cytokines in serum and synovial fluid in rabbits with knee osteoarthritis （KOA）. Methods New Zealand rabbits （n = 32 ） were randomly divided into normal group （n = 5 ）, model group, treatment group （ treated with ultrasound for enhancing permeation of Tongluo Zhitong Tang） and control group （treated with ultrasound for enhancing permeation of saline, each n = 9 ）. The model of KOA was established by injection of 1.6% papain solution in knee joint cavity. After successful modeling confirmed by pathological sections, treatment group and control group were given intervention treatment for 3 weeks. The samples of serum and synovial fluid were collected after the treatment for one week. The content of cytokines in serum and synovial fluid was detected by using radioimmunoassay. At the same time all rabbits were executed, synovial tissue and articular cartilage were taken and stained with HE staining and observed with microscope. Mankin＇ s histological scores were recorded. Results The re-sults of Mankin＇ s histological scoring showed that there were significant difference in scores of synovial tissue （ P 〈 0.05 ）, and no significant difference in scores of cartilage cells （ P 〉 0.05 ） between treatment groups and model group. There wag no significant difference in scores of synovial tissue and cartilage cells between control group and model group （ P 〉 0. 05 ）. Compared with normal group, the content of eytokines in serum and synovial fluid increased significantly in model group （ P 〈 O. 05 ）. Compared with model group, the content of eytokines in synovial fluid decreased significantly in treatment group （ P 〈 0. 05 ）, but had no statistical significance in serum. Conclusion The therapeutic mechanism of low- frequency ultrasound for enhancing permeation of Tongluo Zhitong Tang in treatment of KOA may be related to the inhibition of the generation of eytokines in synovial fluid and relief of knee inflammatory response through slowing the inflammatory response of synovial cells.