医学临床研究
醫學臨床研究
의학림상연구
JOURNAL OF CLINICAL RESEARCH
2014年
4期
740-742,745
,共4页
关节炎 ,类风湿%NF-κB%赤芍
關節炎 ,類風濕%NF-κB%赤芍
관절염 ,류풍습%NF-κB%적작
Arthritis,Rheumatoid%NF-kappa B%RADIX PAEONIAE RUBRA
【目的】探讨赤芍801作用后的类风湿关节炎（rheumatoid arthritis ，RA ）患者成纤维样滑膜细胞（Fibroblast-like synoviocytes ，FLS）p65和p50蛋白在细胞内定位及表达变化。【方法】RA患者关节滑液经原代培养分离出FLS ，分别用浓度为64μg/mL、4μg/mL和0μg/mL的赤芍801作用24 h后，先采用细胞免疫化学方法对p65和p50在细胞内表达定位进行分析，再提取核蛋白，采用Western blotting检测p65和p50的表达。【结果】细胞免疫化学结果表明0μg/mL赤芍801刺激时p65和p50蛋白主要表达于RA患者FLS细胞浆中；与对照组相比，4μg/m L赤芍801浓度刺激24 h后，p65和p50表达无变化（ P ＞0．05），而64μg/m L赤芍刺激24 h后，p65和p50在细胞核中的表达明显上升，与对照组相比差异有统计学意义（ P ＜0．05）。Western blotting结果表明，p65和p50均在RA患者 FLS核蛋白抽提物中被检出，与对照组相比，在4μg/m L赤芍801刺激24 h对p65和p50表达无影响（ P ＞0．05）；当赤芍801浓度增加到64μg/m L时，p65和p50表达显著增加（ P ＜0．05）。【结论】赤芍801可调节RA患者FLS p65和p50从细胞质向细胞核转移，并诱导其表达上调。
【目的】探討赤芍801作用後的類風濕關節炎（rheumatoid arthritis ，RA ）患者成纖維樣滑膜細胞（Fibroblast-like synoviocytes ，FLS）p65和p50蛋白在細胞內定位及錶達變化。【方法】RA患者關節滑液經原代培養分離齣FLS ，分彆用濃度為64μg/mL、4μg/mL和0μg/mL的赤芍801作用24 h後，先採用細胞免疫化學方法對p65和p50在細胞內錶達定位進行分析，再提取覈蛋白，採用Western blotting檢測p65和p50的錶達。【結果】細胞免疫化學結果錶明0μg/mL赤芍801刺激時p65和p50蛋白主要錶達于RA患者FLS細胞漿中；與對照組相比，4μg/m L赤芍801濃度刺激24 h後，p65和p50錶達無變化（ P ＞0．05），而64μg/m L赤芍刺激24 h後，p65和p50在細胞覈中的錶達明顯上升，與對照組相比差異有統計學意義（ P ＜0．05）。Western blotting結果錶明，p65和p50均在RA患者 FLS覈蛋白抽提物中被檢齣，與對照組相比，在4μg/m L赤芍801刺激24 h對p65和p50錶達無影響（ P ＞0．05）；噹赤芍801濃度增加到64μg/m L時，p65和p50錶達顯著增加（ P ＜0．05）。【結論】赤芍801可調節RA患者FLS p65和p50從細胞質嚮細胞覈轉移，併誘導其錶達上調。
【목적】탐토적작801작용후적류풍습관절염（rheumatoid arthritis ，RA ）환자성섬유양활막세포（Fibroblast-like synoviocytes ，FLS）p65화p50단백재세포내정위급표체변화。【방법】RA환자관절활액경원대배양분리출FLS ，분별용농도위64μg/mL、4μg/mL화0μg/mL적적작801작용24 h후，선채용세포면역화학방법대p65화p50재세포내표체정위진행분석，재제취핵단백，채용Western blotting검측p65화p50적표체。【결과】세포면역화학결과표명0μg/mL적작801자격시p65화p50단백주요표체우RA환자FLS세포장중；여대조조상비，4μg/m L적작801농도자격24 h후，p65화p50표체무변화（ P ＞0．05），이64μg/m L적작자격24 h후，p65화p50재세포핵중적표체명현상승，여대조조상비차이유통계학의의（ P ＜0．05）。Western blotting결과표명，p65화p50균재RA환자 FLS핵단백추제물중피검출，여대조조상비，재4μg/m L적작801자격24 h대p65화p50표체무영향（ P ＞0．05）；당적작801농도증가도64μg/m L시，p65화p50표체현저증가（ P ＜0．05）。【결론】적작801가조절RA환자FLS p65화p50종세포질향세포핵전이，병유도기표체상조。
[Objective] To explore the location and expression changes of p65 and p50 protein in fibroblast-like synoviocyte(FLS) in patients with rheumatoid arthritis (RA) after Radix paeoniae rubra 801(P801) treat-ment .[Methods] FLS was isolated from synovial fluid through primary culture ,and then treated with 64 ,4 and 0μg/mL P801 for 24h .Immunocytochemistry method was used to analyze the location of p 65 and p50 in cells ,and then nuclear protein was drawn .Western blotting was used to detect the expression of p65 and p50 .[Results] Immunocytochemistry showed that the p65 and p50 were mainly expressed in cytoplasm of RA FLS treated by 0μg/mL P8010 .Compared with control group ,the expression of p65 or p50 had no change 24h af-ter 4μg/mL P801 treatment( P >0 .05) .The expression of p65 and p50 in nucleus 24h after 64μg/mL P801 treatment ,and there was significant difference between treatment group and control group ( P<0 .05) .West-ern blotting results showed that both p65 and p50 were detected in nucleus extract of RA FLS .Compared with control group ,the treatment with 4μg/mL P801 for 24h had no influence on the expression of p65 and p50( P>0 .05) .When the concentration of P801 reached 64μg/mL ,the expression of p65 and p50 increased signifi-cantly( P<0 .05) .[Conclusion]P801 can regulate the migration of p65 and p50 of RA FLS from cytoplasm to cell nuclear ,and induce their up-regulation .
