富血小板血浆联合仙灵骨葆胶囊治疗大鼠膝骨关节炎的应用研究

Application of platelet-rich plasma combined with Xianling Gubao Capsule in treatment of rats with knee osteoarthritis

摘要

目的 探讨富血小板血浆（platelet-rich plasma，PRP）联合仙灵骨葆胶囊对大鼠骨关节炎（osteoarthritis，OA）的治疗效果及相关因子水平变化。 方法 使用前交叉韧带横断手术构建大鼠骨关节炎模型，模型建立之后将大鼠随机分为对照组（n=10）、模型组（n=10）、仙灵骨葆胶囊组（n=10）、PRP组（n=10）和仙灵骨葆胶囊联合PRP组（n=10）。干预结束后，采用酶联免疫吸附测定（enzyme linked immunosorbent assay，ELISA）检测大鼠血清中白细胞介素-1β（interleukin-1 beta，IL-1β）和肿瘤坏死因子-α（tumor necrosis factor alpha，TNF-α）的水平；取大鼠膝关节软骨，通过qPCR和Western blot检测MMP3、MMP13、Col2、Aggrecan和p-p38基因和蛋白的表达水平；Micro-CT评估大鼠膝关节周围骨变化情况；HE染色和SF染色观察大鼠膝关节软骨变化。 结果 ELISA结果显示，PRP联合仙灵骨葆胶囊治疗后显著降低了大鼠血清中IL-1β和TNF-α等炎性因子水平，差异有统计学意义（P<0.05）。qPCR和Ｗestern blot结果显示PRP联合仙灵骨葆胶囊治疗明显降低大鼠膝关节软骨中MMP3、MMP13、p-p38的表达降低，增加Col2和Aggrecan的表达，差异有统计学意义（P<0.05）。Micro-CT、HE染色和SF染色结果显示，PRP联合仙灵骨葆胶囊组大鼠膝关节软骨退变在一定程度得到改善，差异有统计学意义（P<0.05）。 结论 PRP联合仙灵骨葆胶囊能有效治疗大鼠OA，其机制可能是通过调控p38MAPK信号通路来实现的。

Abstract

Objective To investigate the therapeutic effect of platelet-rich plasma（PRP） combined with Xianling Gubao Capsule on rats with osteoarthritis（OA） and the changes in the levels of related factors. Methods Anterior cruciate ligament transection was performed to establish a rat model of OA，and after modeling，the rats were randomly divided into control group，model group，Xianling Gubao Capsule group，PRP group，and Xianling Gubao Capsule+PRP group，with 10 rats in each group. After intervention，ELISA was used to measure the serum levels of IL-1β and TNF-α； knee articular cartilage was collected，and qPCR and Western blot were used to measure the mRNA and protein expression levels of MMP3，MMP13，Col2，Aggrecan，and p-p38； Micro-CT was used to assess the changes in bones around the knee joint； HE staining and SF staining were used to observe the changes in knee articular cartilage. Results ELISA showed that PRP combined with Xianling Gubao Capsule significantly reduced the serum levels of the inflammatory factors such as IL-1β and TNF-α（P<0.05）. The results of qPCR and Western blot showed that PRP combined with Xianling Gubao Capsule significantly reduced the expression levels of MMP3，MMP13，and p-p38 and increased the expression levels of Col2 and Aggrecan in knee articular cartilage（P<0.05）. Micro-CT，HE staining，and SF staining showed a certain degree of improvement in knee articular cartilage degeneration in the Xianling Gubao Capsule+PRP group （P<0.05）. Conclusion PRP combined with Xianling Gubao Capsule exerts a therapeutic effect on rats with OA，possibly by regulating the p38MAPK signaling pathway.

关键词

骨关节炎 / 富血小板血浆 / 仙灵骨葆胶囊 / p38丝裂原活化蛋白激酶

Key words

osteoarthritis / platelet-rich plasma / Xianling Gubao Capsule / p38 mitogen-activated protein kinase

基金资助

广东省中医药局科研资助项目(20241110)