CAJ | 학술논문

目的：探讨类风湿关节炎（ RA）血瘀病机的分子基础。方法用肿瘤坏死因子－α（ TNF－α）诱导RA成纤维样滑膜细胞MH7A，构建RA体外模型；采用MTS法检测甲氨蝶呤（MTX）及青藤碱（SIN）对MH7A细胞增殖的影响；采用Western blot法检测不同浓度MTX及SIN对α辅肌动蛋白－1（ACTN1）、血栓素合酶（TxAS）、环氧酶－2（COX－2）蛋白表达的影响，并检测ACTN1、TxAS蛋白在RA不同证型滑膜组织中的表达。结果 MTS结果显示，50、200、400μg／mL的MTX及SIN作用24、48、72 h均能抑制MH7A细胞的增殖；Western blot结果显示， MTX及SIN均能抑制ACTN1、TxAS、COX－2的表达，且在TNF－α诱导模型中更显著；MH7A细胞中ACTN1、Tx-AS、COX－2的表达趋势具有一致性；寒湿痹阻证、肝肾不足证滑膜组织中ACTN1、TxAS的表达均显著高于湿热痹阻证。结论 TxAS可以被经典抗风湿药物MTX和SIN所调节，且与RA滑膜细胞增殖与侵袭的标志物ACTN1表达一致，说明TxAS可能是RA血瘀病机的分子基础之一。
목적：탐토류풍습관절염（ RA）혈어병궤적분자기출。방법용종류배사인자－α（ TNF－α）유도RA성섬유양활막세포MH7A，구건RA체외모형；채용MTS법검측갑안접령（MTX）급청등감（SIN）대MH7A세포증식적영향；채용Western blot법검측불동농도MTX급SIN대α보기동단백－1（ACTN1）、혈전소합매（TxAS）、배양매－2（COX－2）단백표체적영향，병검측ACTN1、TxAS단백재RA불동증형활막조직중적표체。결과 MTS결과현시，50、200、400μg／mL적MTX급SIN작용24、48、72 h균능억제MH7A세포적증식；Western blot결과현시， MTX급SIN균능억제ACTN1、TxAS、COX－2적표체，차재TNF－α유도모형중경현저；MH7A세포중ACTN1、Tx-AS、COX－2적표체추세구유일치성；한습비조증、간신불족증활막조직중ACTN1、TxAS적표체균현저고우습열비조증。결론 TxAS가이피경전항풍습약물MTX화SIN소조절，차여RA활막세포증식여침습적표지물ACTN1표체일치，설명TxAS가능시RA혈어병궤적분자기출지일。
Objective To investigate the molecular basis of pathogenesis of blood stasis in rheumatoid arthritis (RA).Methods Tumor necrosis factor alpha (TNF-α) was used to induce fibroblast -like synoviocytes MH7A of RA, so as to establish an in vitro model of RA.By means of MTS method, the effect of methotrexate (MTX) and sinome-nine ( SIN) on proliferation of MH7A cells was detected .The effect of different concentrations of MTX and SIN on expres-sion of alpha actinin-1 (ACTN1), thromboxane synthase (TxAS), cyclooxygenase -2 (COX-2) was detected using Western blot .The expression of ACTN 1 and TxAS in synovial tissues of different syndrome types of RA was also detected . Results MTS showed that both MTX and SIN, at 50, 200, 400 μg/mL, could inhibit the proliferation of MH7A, under treatment of 24, 48 and 72 h.The results of Western blot showed that expression of ACTN 1, TxAS and COX-2 was in-hibited by MTX and SIN , which was more remarkable in TNF -α-induced model .And the trends of expression of ACTN1, TxAS and COX-2 were consistent in the MH7A cell line.Expressions of ACTN1 and TxAS in synovial tissues of types Hanshizubi and Ganshenbuzu were significantly higher than those of type Shirezubi .Conclusion TxAS can be regu-lated by classical anti-rheumatic agents MTX and SIN , and its expression is consistent with that of ACTN 1, the marker of synovial cell proliferation and invasion in RA .These indicate that TxAS may be part of the molecular basis to the patho-genesis of blood stasis in RA .