[1]朱俊卿,王然,陈世贤,等.白细胞介素-22促进类风湿关节炎成纤维化膜细胞的增殖[J].南方医科大学学报,2017,(12):1570.
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白细胞介素-22促进类风湿关节炎成纤维化膜细胞的增殖()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2017年12期
页码:
1570
栏目:
出版日期:
2017-12-20

文章信息/Info

Title:
Interleukin-22 promotes proliferation of fibroblast-like synoviocytes from patients with rheumatoid arthritis by inducing STAT3 phosphorylation
作者:
朱俊卿王然陈世贤赵迪李娟
关键词:
类风湿关节炎成纤维滑膜细胞白细胞介素22细胞信号通路
Keywords:
rheumatoid arthritis fibroblast-like synoviocytes interleukin-22 signaling pathway
摘要:
目的通过白细胞介素-22(IL-22)干预类风湿关节炎(RA)成纤维滑膜细胞(FLS)以期明确IL-22 促进其增殖的机制。 方法无菌条件下获取的RA滑膜组织,应用组织块分离法培养RA-FLS,含10% FBS的DMEM培养基进行细胞传代培养,并对 第4~5代FLS应用vimentin/CD68进行免疫组化法鉴定;不同浓度的IL-22干预FLS培养24、48、72 h后,MTT法检测细胞增殖 程度;联合IL-22和/或AG490干预FLS,于冰面上用含蛋白酶抑制剂的细胞裂解液获取细胞总蛋白,再经超声、离心提取、蛋白 定量后,Western blot法检测STAT3、ERK1/2、P38蛋白及其磷酸化蛋白的表达,统计分析采用SPSS20.0软件进行,P<0.05为差 异有统计学意义。结果不同浓度IL-22干预FLS的增殖均较对照组呈梯度增高变化,差异具有统计学意义(P<0.05)。随着IL- 22干预时间的延长,STAT3总蛋白表达相对灰度值的整体水平差异无统计学意义(P=0.68),但是其磷酸化蛋白表达相对灰度值 整体水平差异具有统计学意义(P<0.001),且在干预的不同时间点STAT3磷酸化蛋白相对表达量较基线0 h时均显著升高,且 差异具有统计学意义(P<0.001)。IL-22 干预后的ERK1/2 和P38 总蛋白及其磷酸化蛋白表达相对灰度值差异均无统计学意 义(P>0.05)。联合50 ng/mL IL-22和100 μmol/LAG490干预FLS不同时间后,细胞的增殖程度较单用IL-22 干预均显著降低 (P<0.01)。结论IL-22以浓度依赖方式促进RA成纤维化膜细胞STAT3蛋白磷酸化所介导的细胞增殖,而非ERK1/2和P38蛋 白所介导的信号通路。
Abstract:
Objective To clarify the mechanism by which interleukin-22 (IL-22) promotes the proliferation of fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA). Methods FLS were isolated from the synovial tissues of patients with RA and identified by immunohistochemistry for vimentin/CD68. The cells were subcultured and incubated with different concentrations of IL-22 for 24, 48, or 72 h, and their proliferation was examined using MTT assay. After treatment of the cells with IL-22 and AG490, alone or in combination, the expressions of the total and phosphorylated proteins of STAT3, ERK1/2 and P38 were detected with Western blotting. Results IL-22 significantly increased the proliferation of FLS in a dose-dependent manner (P<0.05). The total protein of STAT3 in the cells showed no significant changes with extended time of IL-22 treatment (P=0.68), but the expression of phosphorylated STAT3 protein increased significantly (P<0.001). The total and phosphorylated proteins of ERK1/2 and P38 underwent no significant changes after IL-22 treatment (P>0.05). A combined treatment with 50 ng/mL IL-22 and 100 μmol/L AG490 resulted in a significant decrease in the proliferation of FLS as compared with IL-22 treatment alone (P<0.01). Conclusion IL-22 can dose-dependently promote the proliferation of FLS from patients with RA by inducing phosphorylation of STAT3 protein but not through ERK1/2 or P38 signal pathway.

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更新日期/Last Update: 1900-01-01