中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2014年
2期
87-90,后插1
,共5页
邹婵娟%莫颖倩%朱浪静%韦秀宁%郑东辉%戴冽
鄒嬋娟%莫穎倩%硃浪靜%韋秀寧%鄭東輝%戴冽
추선연%막영천%주랑정%위수저%정동휘%대렬
淋巴瘤,B细胞%关节炎,类风湿%破骨细胞
淋巴瘤,B細胞%關節炎,類風濕%破骨細胞
림파류,B세포%관절염,류풍습%파골세포
Lymphoma,B-cell%Arthritis,rheumatoid%Osteoclasts
目的 探讨B细胞淋巴瘤因子(Bcl)6在RA破骨细胞形成及活化过程中的动态变化及意义.方法 体外分离培养RA成纤维样滑膜细胞(FLS),加入巨噬细胞集落刺激因子,与健康人PBMCs共培养0、7、14、21 d,细胞免疫荧光、Real-time PCR分别检测破骨细胞及前体Bcl6蛋白和mRNA表达;抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞,甲苯胺蓝染色观察骨吸收陷窝情况.多组间均数比较采用Kruskal-Wallis H检验和Bonferroni法进行统计学分析.结果 ①健康人PBMCs中Bcl6蛋白表达弱,主要表达于细胞核;培养7d时,部分PBMCs分化成巨噬细胞及少量TRAP(+)多核破骨细胞[(664±78)个/孔],破骨细胞及其前体总体Bcl6蛋白表达升高;14 d时总体Bcl6蛋白表达继续升高;21 d时PBMCs基本被诱导分化为多核破骨细胞,后者细胞核中表达的Bcl6蛋白减少,总体Bcl6蛋白表达亦降低.②RA-FLS与PBMCs共培养至7d时,破骨细胞及前体总体Bcl6 mRNA量较0d时升高,但差异无统计学意义(x2=3.429,P＞0.05);至14 d时Bcl6 mRNA量较0d时显著升高(x2=5.333,P=0.045);至21 d时则较14 d时明显降低(x2=6.023,P=0.038).结论 Bcl6可能参与调控RA破骨细胞的形成及活化以及PBMCs分化为巨噬细胞的炎症过程,相关机制值得进一步研究.
目的 探討B細胞淋巴瘤因子(Bcl)6在RA破骨細胞形成及活化過程中的動態變化及意義.方法 體外分離培養RA成纖維樣滑膜細胞(FLS),加入巨噬細胞集落刺激因子,與健康人PBMCs共培養0、7、14、21 d,細胞免疫熒光、Real-time PCR分彆檢測破骨細胞及前體Bcl6蛋白和mRNA錶達;抗酒石痠痠性燐痠酶(TRAP)染色鑒定破骨細胞,甲苯胺藍染色觀察骨吸收陷窩情況.多組間均數比較採用Kruskal-Wallis H檢驗和Bonferroni法進行統計學分析.結果 ①健康人PBMCs中Bcl6蛋白錶達弱,主要錶達于細胞覈;培養7d時,部分PBMCs分化成巨噬細胞及少量TRAP(+)多覈破骨細胞[(664±78)箇/孔],破骨細胞及其前體總體Bcl6蛋白錶達升高;14 d時總體Bcl6蛋白錶達繼續升高;21 d時PBMCs基本被誘導分化為多覈破骨細胞,後者細胞覈中錶達的Bcl6蛋白減少,總體Bcl6蛋白錶達亦降低.②RA-FLS與PBMCs共培養至7d時,破骨細胞及前體總體Bcl6 mRNA量較0d時升高,但差異無統計學意義(x2=3.429,P＞0.05);至14 d時Bcl6 mRNA量較0d時顯著升高(x2=5.333,P=0.045);至21 d時則較14 d時明顯降低(x2=6.023,P=0.038).結論 Bcl6可能參與調控RA破骨細胞的形成及活化以及PBMCs分化為巨噬細胞的炎癥過程,相關機製值得進一步研究.
목적 탐토B세포림파류인자(Bcl)6재RA파골세포형성급활화과정중적동태변화급의의.방법 체외분리배양RA성섬유양활막세포(FLS),가입거서세포집락자격인자,여건강인PBMCs공배양0、7、14、21 d,세포면역형광、Real-time PCR분별검측파골세포급전체Bcl6단백화mRNA표체;항주석산산성린산매(TRAP)염색감정파골세포,갑분알람염색관찰골흡수함와정황.다조간균수비교채용Kruskal-Wallis H검험화Bonferroni법진행통계학분석.결과 ①건강인PBMCs중Bcl6단백표체약,주요표체우세포핵;배양7d시,부분PBMCs분화성거서세포급소량TRAP(+)다핵파골세포[(664±78)개/공],파골세포급기전체총체Bcl6단백표체승고;14 d시총체Bcl6단백표체계속승고;21 d시PBMCs기본피유도분화위다핵파골세포,후자세포핵중표체적Bcl6단백감소,총체Bcl6단백표체역강저.②RA-FLS여PBMCs공배양지7d시,파골세포급전체총체Bcl6 mRNA량교0d시승고,단차이무통계학의의(x2=3.429,P＞0.05);지14 d시Bcl6 mRNA량교0d시현저승고(x2=5.333,P=0.045);지21 d시칙교14 d시명현강저(x2=6.023,P=0.038).결론 Bcl6가능삼여조공RA파골세포적형성급활화이급PBMCs분화위거서세포적염증과정,상관궤제치득진일보연구.
Objective To investigate the dynamic expression and significance of B cell lymphoma (Bcl) 6 in fibroblast-like synoviocytes (FLS) induced osteoclast differentiation and activation in rheumatoid arthritis (RA) patients.Methods RA-FLS were co-cultured with peripheral blood monocytes (PBMCs) from healthy volunteers in the medium containing M-CSF.Bcl6 protein and mRNA in osteoclasts and their precursors were determined by immunofluorescence and Real-time PCR at day 0,7,14 and 21,respectively.Osteoclasts were identified by tartrate-resistant acid phosphatase (TRAP) staining.Bone resorption activity of osteoclasts was determined by bone slices stained with toluidine blue.Kruskal-Wallis H and Bonferroni were used for statistical analysis.Results ① Immunofluorescence staining and TRAP staining showed that Bcl6 protein was mainly expressed in the nuclei of PBMCs.After co-cultured with RA-FLS for 7 days,some PBMCs differentiated into macrophages and a few differentiated to TRAP-positive multinucleated osteoclasts,and the total Bcl6 protein expression in osteoclasts and their precursors were increased.At day 14,the total Bcl6 protein expression was increased further.At day 21,the Bcl6 protein expression in nuclei of osteoclasts was decreased while PBMCs were differentiated into osteoclasts,and total Bcl6 protein expression was decreased.②Real-time PCR showed that Bcl6 mRNA expression in osteoclasts and their precursors at day 7 tended to increase than that at day 0 (x2=3.429,P＞0.05).At day 14 after co-cultured with RA-FLS,Bcl6 mRNA expression in osteoclasts and their precursors was significantly higher than that at day 0 (x2=5.333,P=0.045).At day 21,the expression of Bcl6 mRNA was significantly lower than that at day 14 (x2=6.023,P=0.038).Conclusion Bcl6 may be involved in osteoclast differentiation and activation,and may play a role in the inflammatory status in the process of differentiation from PBMCs to macrophages.Further studies are needed to establish the mechanisms.
