调节性T细胞对肝星状细胞增殖及透明质酸分泌的影响

陆小蒟; 陈永平; 阳韬; 程瑗; 金晓芝; 申春燕; 刘俊; 王春莹; 王晓东; 林镯

调节性T细胞对肝星状细胞增殖及透明质酸分泌的影响

温州医学院附属第一医院

基金项目:

浙江省自然科学基金资助项目(Y207464)；浙江省教育厅项目(Y201009942)；中国肝炎防治基金会王宝恩肝纤维化研究基金(20080017)；

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中图分类号: R575.1
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出版历程
- 出版日期: 2012-01-20

The influence of regulatory T cells in hepatic stellate cell proliferation and HA secretion

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摘要

摘要: 目的研究CD4+CD25+调节性T细胞（Treg）对肝星状细胞（HSC）增殖及透明质酸（HA）分泌的影响。方法 MASC法分离慢性乙型肝炎患者血中CD4+CD25+Treg细胞;传代培养人HSC LX-2（空白组）,不同比例CD4+CD25+Treg细胞与HSC直接共培养（共培养组）,不同剂量的重组人转化生长因子（TGF）β1和HSC共培养（对照组）;使用CCK-8法检测HSC增殖、放射免疫法检测HA分泌情况。组间比较采用单因素方差分析。结果共培养组与空白对照组（0.6623±0.02702）相比,在比例为2.5∶1时HSC增殖最为明显（0.6836±0.00560）,比例为1∶1、5∶1及7.5∶1时HSC增殖均明显减少,且随着比例升高其增殖水平逐渐降低,各组与空白对照组相比有统计学差异（F=11.421,5.272,84.514,490.639;均P<0.05）。不同浓度TGFβ1对照组的HSC均比空白对照组（0.3883±0.00189）细胞增殖明显（0.6720±0.00753,0.6773±0.00585,0.5373±0.00387）,并于8 ng/μl时增殖达到高峰...
- T淋巴细胞 /
- 肝星状细胞 /
- 透明质酸,细胞增殖
Abstract: Objective To investigate the effect of CD4+CD25+ regulatory T cells (Treg) in the proliferation of hepatic stellate cells (HSC) and the secretion of Hyaluronic Acid (HA) .Methods By using magnetic activated cell sorter (MRCS) , Treg cells were isolated from peripheral blood of chronic hepatitis B patients The hepatic stellate cells (HSC) cultured alone were used as blank group.The LX-2 were co-cultured with Treg cells in vitro as co-culture group;adding TGFβ1 to HSC co-cultured as control group.CCK-8 method for the detection of the proliferation of HSC, ELISA analysis for the detection of HA secretion was performed.One-way analysis of variance was used for the comparison.Results Compared with the blank group (0.6623±0.02702) the proliferation of HSC of the co-culture group was significantly higherand the most prominent effect was 2.5∶ 1 (0.6836±0.00560) , while the proliferation of HSC decreased at 1∶ 1、5∶ 1 and 7.5∶ 1, and the discrepancy between co-culture subgroups and blank group had statistical significance (F=11.421, 5.272, 84.514, 490.639;P<0.05) .Compared with blank group, the proliferation of HSCs of different concentrations of TGFβ1 control subgroup increased significantly (0.6720±0.00753, 0.6773±0.00585, 0.5373±0.00387) and reached a peak at 8 ng/μl, and the discrepancy between control subgroups and blank group had statistical significance (F=196.175, 799.624, 236.461;P<0.01) .Linear correlation analysis showed concentrations of HA and the proliferation of HSC in co-culture group was positively correlated with control group (r=0.661, 0.871;P<0.01) .Conclusion CD4+CD25+ Treg cells could accelerate the cultivation of HSCs proliferation and HA secretion.And, in vitro, we confirmed CD4+CD25+ Treg cells play an important role in the process of chronic hepatitis B.
- T-lymphocytes /
- hepatic stellate cells /
- hyaluronic acid

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