CAJ | 학술논문

目的：探讨青藤碱（Sinomenine，SIN）对TLR信号转导通路及髓样分化因子88（Myeloid differentiation factor 88，MyD88）、肿瘤坏死因子受体相关因子6（ Tumor necrosis factor receptor associated factor－6，TRAF－6）表达的影响，阐明SIN抑制类风湿关节炎（ Rheumatoid arthritis，RA）成纤维样滑膜细胞（ Fibroblast－like synoviocytes，FLS）增生导致RA软骨和软骨下骨破坏造成关节畸形的作用机制。方法：体外培养RA－FLS细胞，分为对照组与（0．125、0．25、0．5、1 mmol／L） SIN组，通过检测各组细胞内碱性磷酸酶（ ALP）活性，确定体外用药最佳药物浓度；CCK－8法检测0．5 mmol／L SIN组的细胞增殖率；荧光定量PCR法测定对照组与0．5 mmol／L SIN组MyD88和TRAF－6基因表达；Western blot法检测对照组与0．5 mmol／L SIN组MyD88和TRAF－6蛋白表达。结果：SIN各组ALP活性均低于对照组，其中以0．5 mmol／L SIN组的ALP活性为最低（P＜0．01）。 CCK－8法检测，0．5 mmol／L SIN组RA－FLS细胞增殖率明显低于对照组（ P＜0．01），SIN诱导第4天细胞增殖率最高，进入平台期，之后细胞的增殖率开始下降。荧光定量PCR和Western blot法检测，0．5 mmol／L SIN组的MyD88和TRAF－6基因及蛋白表达明显低于对照组，差异有统计学意义（P＜0．01）。结论：SIN通过对TLR信号转导通路的影响有效地抑制RA－FLS细胞内MyD88和TRAF－6的表达，这可能是其治疗RA防止软骨和软骨下骨破坏造成关节畸形发生的重要分子机制之一。
목적：탐토청등감（Sinomenine，SIN）대TLR신호전도통로급수양분화인자88（Myeloid differentiation factor 88，MyD88）、종류배사인자수체상관인자6（ Tumor necrosis factor receptor associated factor－6，TRAF－6）표체적영향，천명SIN억제류풍습관절염（ Rheumatoid arthritis，RA）성섬유양활막세포（ Fibroblast－like synoviocytes，FLS）증생도치RA연골화연골하골파배조성관절기형적작용궤제。방법：체외배양RA－FLS세포，분위대조조여（0．125、0．25、0．5、1 mmol／L） SIN조，통과검측각조세포내감성린산매（ ALP）활성，학정체외용약최가약물농도；CCK－8법검측0．5 mmol／L SIN조적세포증식솔；형광정량PCR법측정대조조여0．5 mmol／L SIN조MyD88화TRAF－6기인표체；Western blot법검측대조조여0．5 mmol／L SIN조MyD88화TRAF－6단백표체。결과：SIN각조ALP활성균저우대조조，기중이0．5 mmol／L SIN조적ALP활성위최저（P＜0．01）。 CCK－8법검측，0．5 mmol／L SIN조RA－FLS세포증식솔명현저우대조조（ P＜0．01），SIN유도제4천세포증식솔최고，진입평태기，지후세포적증식솔개시하강。형광정량PCR화Western blot법검측，0．5 mmol／L SIN조적MyD88화TRAF－6기인급단백표체명현저우대조조，차이유통계학의의（P＜0．01）。결론：SIN통과대TLR신호전도통로적영향유효지억제RA－FLS세포내MyD88화TRAF－6적표체，저가능시기치료RA방지연골화연골하골파배조성관절기형발생적중요분자궤제지일。
Objective: To investigate SIN ( Sinomenine) for TLR signal transduction pathway and MyD88 ( MyeloidDifferentiation Factor 88), TRAF-6 ( Tumor necrosis factor receptor associated factor-6) expression, clarifying SIN inhibit RA(Rheumatoid arthritis)-FLS(Fibroblast-like synoviocytes)proliferation leads to joint deformity of RA cartilage and subchondral bonedestruction caused by the role of mechanisms.Methods: RA-FLS cells for vitro were divided into a control group and(0.125,0.25,0.5,1 mmol/L)SIN group,within each group were detected by alkaline phosphatase(ALP)activity,to determine the best concentrationfor vitro drug;detect 0.5 mmol/L SIN group in CCK-8 method to detect cell proliferation rate;fluorescence quantitative PCR methodMyD88 SIN group and control group with 0.5 mmol/L TRAF-6 gene expression;Western blot method to detect MyD88 SIN group andcontrol group with 0.5 mmol/L TRAF-6 protein expression.Results: SIN the ALP activity of the lower than the control group,with theminimum ALP activity of 0.5 mmol/L SIN group(P<0.01).CCK 8 method,0.5 mmol/L RA-FLS cell proliferation rate SIN group wasobviously lower than the control group(P<0.01),SIN induce cell proliferation rate was highest,4 days into the plateau,after cell proliferationrate began to fall.Fluorescence quantitative PCR and Western blot method to detect,0.5 mmol/L SIN group of MyD88 andTRAF-6 gene and protein expression significantly lower than the control group,the difference was statistically significant(P<0.01). Conclusion: The SIN of TLR signalling pathways through effectively suppress the influence of RA-FLS MyD88 in the cell and theexpression of TRAF-6,this could be a treatment of RA prevent cartilage and subchondral bone damage cause joint deformity happened one of the important molecular mechanism.