昆山合剂对类风湿关节炎合并贫血患者滑膜成纤维细胞增殖及MyD88、IL-6表达的研究

Study on Proliferation of Fibroblast-like Synoviocytes and MyD88 and IL-6 Expression among Rheumatoid Arthritis Patients with Anemia Treated with Kunshan Mixture

探讨昆山合剂(昆明山海棠、山血丹组成)对体外培养的类风湿关节炎(RA)致慢性病贫血患者滑膜成纤维细胞(FLS)增殖以及人类髓样分化蛋白88(MyD88)mRNA及其下游因子IL-6表达的影响和机制。方法:制备甲氨蝶呤和昆山合剂高、中、低剂量组的家兔含药血清;离体培养RA-FLS,取第3至5代细胞加入各组含药血清干预,MTT法检测各含药血清组和对照组RA-FLS的增殖情况,RT-PCR检测含药血清对该细胞MyD88 mRNA表达的影响。结果:干预24、48、72 h之后,与空白组比较,各浓度组的昆山合剂组和甲氨蝶呤含药血清作用均能明显抑制RA-FLS增殖(P<0.05),对脂多糖诱导的MyD88 mRNA高表达也有明显抑制作用(P<0.05),与脂多糖诱导组比较,48 h后昆山合剂高剂量组细胞培养上清的IL-6含量明显降低(P<0.05)。结论:昆山合剂含药血清能显著抑制RA-FLS增殖,并可下调脂多糖诱导的TLR4mRNA及其下游炎症因子IL-6的表达,这可能为该方治疗RA的机制之一。

This study was aimed to investigate the effects and mechanisms of serum containing Kunshan Mixture （KSM） on in vitro proliferation of fibroblast-like synoviocytes（FLS） and myeloid differentiation protein 88 （MyD88） mRNA and the expression of its downstream factor IL-6 in rheumatoid arthritis patients with chronic anemia. Rabbit medicated serum containing methotrexate （MTX） and KSM of high-, medium-, and low-dose were pre-pared. In vitro culture of RA-FLS was given. Cells from the 3rd to 5th generation were intervened with medicat-ed serum of different groups. MTT method was used in the detection of RA-FLS proliferation condition among different medicated serum groups and the control group . RT-PCR was used in the detection of MyD88 mRNA ex-pression of these cells. The results showed that compared with the blank control group, after the intervention of 24 , 48 , or 72 h , KSM of different concentrations and MTX medicated serum had obvious inhibitive effects on the proliferation of RA-FLS （P 〈 0.05）. It also had obvious inhibitive effects on the high level expression of LPS-induced MyD88 mRNA （P 〈 0.05）. Compared with the LPS-induced group, the supernatant IL-6 content was ob-viously reduced in the KSM of high-dose group after 48 h （ P 〈 0 . 05 ） . It was concluded that KSM medicated serum can obviously inhibit the proliferation of RA-FLS and downregulate the expression of LPS-induced TLR4mRNA and its downstream inflammatory factor IL-6. It may be one of the mechanisms for RA treatment with this prescription .