化瘀祛湿方对早期膝骨关节炎滑膜成纤维细胞COX―2 mRNA表达影响.docVIP

化瘀祛湿方对早期膝骨关节炎滑膜成纤维细胞COX―2 mRNA表达影响 　　[摘要] 目的 通过观察化瘀祛湿方对早期膝骨关节炎（KOA）患者滑膜成纤维细胞（FLS）中COX-2 mRNA表达的影响，阐释化瘀祛湿方对早期KOA治疗的作用机制。 方法 通过切断膝关节前交叉韧带和内侧副韧带，将成年健康新西兰雄性大耳白兔16只造成KOA模型，并将其随机分为生理盐水组（A组）、补肾活血组（B组）、化瘀祛湿组（C组）、塞来昔布组（D组），造模成功后6周，连续喂药7 d后处死实验兔，静脉穿刺采血，离心后取血清备用。将收集的早期KOA患者滑膜组织，经分离、纯化培养后，使用四代FLS用于实验，依次分别加入含A、B、C、D组药物的DMEM培养基中培养。重复离心后，应用RT-PCR检测四组含药血清干预后早期KOA患者FLS中COX-2 mRNA的表达水平。 结果 早期KOA患者FLS经不同含药血清干预后均能检测到COX-2 mRNA，A组的COX-2 mRNA表达水平最高，与B、C、D组比较差异有高度统计学意义（P 0.05）。 结论 化瘀祛湿方可有效抑制早期KOA患者FLS中COX-2 mRNA的表达，在治疗早期KOA中发挥重要作用。 　　[关键词] 膝骨关节炎；化瘀祛湿方；滑膜成纤维细胞；COX-2 　　[中图分类号] R684.3 [文献标识码] A [文章编号] 1674-4721（2016）08（c）-0040-06 　　[Abstract] Objective Through observing the effect of Removing Blood Stasis and Eliminating Dampness Decoction for the expression of COX-2 mRNA of synovial fibroblasts （FLS） in patients with early knee osteoarthritis （KOA）， to illustrate the mechanism of Removing Blood Stasis and Eliminating Dampness Decoction for the treatment of early KOA. Methods Through cutting off the anterior cruciate ligament and medial collateral ligament of knee joint， 60 healthy adult New Zealand male special rabbits was made into KOA models， and they were randomly divided into normal saline group （group A）， nourishing kidney and activating blood group （group B）， removing blood stasis and eliminating dampness group （group C）， Celecoxib group （group D）. After making models successfully for 6 weeks， the experimental rabbits were put to death after continuous medicine feeding for 7 d， collecting blood through venipuncture， the serum was taken as standby application after centrifugation. The collected synovial tissues of patients with early KOA were taken separation and purification culture， so as to be used in the experiment by the fourth generation of FLS， where were added into DMEM medium including the medicines of group A， B， C， D respectively. After repeated centrifugation， RT-PCR was used to detect the expression of COX-2 mRNA of FLS in the patients with early KOA after interventi