[1]任 洁,周 毅,吴会霞,等.NK-22细胞及IL-22相关功能分子在类风湿关节炎滑膜细胞增殖中的作用[J].南方医科大学学报,2014,(01):20.
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NK-22细胞及IL-22相关功能分子在类风湿关节炎滑膜细胞增殖中的作用()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2014年01期
页码:
20
栏目:
出版日期:
2014-01-15

文章信息/Info

Title:
Role of NK-22 cells and interleukin-22-related molecules in proliferation of fibroblastlike
synoviocytes in patients with rheumatoid arthritis
作者:
任 洁周 毅吴会霞戴桃李朱丽花
关键词:
类风湿关节炎NK-22细胞IL-22STAT3
Keywords:
rheumatoid arthritis natural killer-22 cells interlukin-22 signal transduction and activator of transcription 3
摘要:
目的探讨类风湿关节炎(RA)患者滑液(SF)自然杀伤(NK)细胞NK-22促成纤维样滑膜细胞(FLS)增殖作用及相关信号
通路。方法采用流式细胞术分选6 例RA患者SF 中NK-22 细胞(2×104),体外悬浮培养2 周,佛波酯(20 ng/ml)和离子霉素
(0.5 μmol/L)刺激后ELISA检测NK-22细胞培养上清IL-22浓度。组织块培养法原代培养RAFLS,NK-22细胞培养上清分别作
用于RAFLS24、48、72 h,MTT法检测FLS增殖;同时加入IL-22抗体检测FLS增殖情况。RT-PCR检测rhIL-22 及AG490作用
后RAFLS Stat3 mRNA的表达,Western blot 检测RAFLS p-Stat3 蛋白含量。结果①流式分选2×104NK-22 细胞,细胞纯度>
90%;②PMA和ionomycin 刺激后NK-22 细胞培养上清中IL-22 浓度为1273.42±254.48 pg/ml;③PMA和ionomycin 刺激后
NK-22细胞培养上清作用于RAFLS 24 h、48 h、72 h,可明显刺激RAFLS增殖(P<0.05)。IL-22抗体能明显抑制NKp44+NK细
胞上清诱导的RA FLS增殖(P<0.01);④rhIL-22(1 ng/ml)能诱导RAFLS p-Stat3表达(P<0.05);⑤AG490能抑制rhIL-22诱导的
RAFLS p-Stat3 表达(P<0.05)。结论RA患者SFNK-22 细胞体外可分泌高浓度的IL-22,通过激活STAT3 信号通路进而刺激
RAFLS增殖。
Abstract:
Objective To investigate the role of natural killer-22 (NK-22) cells in the synovial fluid in the proliferation of
fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA) and explore the possible signal pathway involved.
Methods NK-22 cells in the SF of RA patients were sorted by flow cytometry. NK-22 cells were cultured for two weeks and the
purity was detected by flow cytometry before stimulation with 20 ng/ml phorbol 12-myristate 13-acetate and 0.5 μmol/L
ionomycin for 4 h. The level of interleukin-22 (IL-22) in the culture medium supernatant was then measured with ELISA. The
proliferation of FLS in the presence of the culture supernatant of NK-22 cells was assessed with MTT assay at 24, 48 and 72 h,
and the effect of IL-22 antibody on FLS proliferation was also observed. Real-time PCR and Western blotting were used to
detect Stat3 mRNA and p-Stat3 protein levels, respectively, in the FLS exposed to rhIL-22 and AG490. Results NK-22 cells were
successfully sorted by flow cytometry with a purity exceeding 90%. The levels of IL-22 in the supernatant of NKp44+NK cell
culture averaged 1273.42±254.48 pg/ml. The FLS proliferated rapidly 24, 48, and 72 h after the addition of culture supernatant
of NK-22 cells (P<0.05). IL-22 antibody obviously inhibited the proliferation of FLS induced by NK-22 cell culture supernatant
(P<0.05). Exposure of the FLS to rhIL-22 obviously increased cellular Stat3 expression levels, which were significantly lowered
by the addition of AG490 (P<0.05). Conclusion NK-22 cells in the SF of RA patients can produce high concentrations of IL-22 to
promote the proliferation of FLS through the STAT3 signal pathway.

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更新日期/Last Update: 1900-01-01