中国血液净化

中国血液净化 ›› 2018, Vol. 17 ›› Issue (02): 93-98.doi: 10.3969/j.issn.1671-4091.2018.02.005

• 基础研究 • 上一篇    下一篇

舒洛地特对高糖作用下人腹膜间皮细胞炎症及纤维化因子表达影响

边爱淑1,2,胡晔1,杨丽娜1,樊怡1,马健飞1   

  1. 1. 中国医科大学附属第一医院肾内科
    2. 沈阳二四二医院
  • 收稿日期:2017-10-30 修回日期:2017-12-18 出版日期:2018-02-12 发布日期:2018-02-12
  • 通讯作者: 马健飞 majiangfei@medmail.com.cn E-mail:majiangfei@medmail.com.cn
  • 基金资助:

    国家自然科学基金(81370865)

Effect of sulodexide on high glucose induced inflammation and fibrosis in human peritoneal mesothelial cells

  • Received:2017-10-30 Revised:2017-12-18 Online:2018-02-12 Published:2018-02-12

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摘要： 目的研究高糖对人腹膜间皮细胞(human peritoneal mesothelial cells, HPMCs)糖萼组成成分透明质酸(hyaluronic acid,HA)、核心蛋白聚糖(decorin,DCN)和炎症及纤维化因子(白细胞介素6, interleukin-6,IL-6)；转化生长因子-β1,transforming growth factor-β1,TGF-β1)表达的影响，以及观察舒洛地特(sulodexide,SLX)对上述因子表达的影响。方法将HPMCs 随机分为以下各组：1.对照组；2.高糖组：①不同浓度组：1.5%，2.5%，4.25%葡萄糖作用48h；②不同时间组：2.5%葡萄糖
作用不同时间(0，6，12，24，48，72h)；3.高糖+SLX 组：2.5%高糖+不同浓度舒洛地特(50μg/ml，400μg/ml，800μg/ml，1200μg/ml)作用48h。ELISA 法检测各组上清液中HA、DCN、IL-6 及TGF-β1 的蛋白表达情况。结果1.与对照组相比，不同浓度的高糖作用下HPMCs HA 表达增多[(172.074±3.223)pg/ml比(147.000 ± 1.667)pg/ml，F=20.410，P=0.036]，DCN 表达增多[(2.435 ± 0.150)ng/ml 比(1.021 ±0.138)ng/ml，F=219.555，P＜0.001]，IL-6 表达增多[(67.981± 1.655ng/l) 比(45.637±1.143ng/l),F=160.674, P＜0.001]，TGF- β1 表达增多[(204.691 ± 2.829)ng/l 比(112.716 ± 7.484)ng/l，F=1692.284,P＜0.001]；不同时间的高糖作用下HPMCs HA 表达增多[(205.986 ± 2.746)pg/ml 比(147.000 ± 1.667)pg/ml，F=1225.533，P＜0.001]，DCN 表达增多[(1.422 ± 0.134)ng/ml 比(1.021 ±0.138)ng/ml，F=112.124，P=0.001]，IL-6 表达增多[(68.078±2.944)ng/l 比(45.637±1.143)ng/l，F=69.690，P＜0.001]，TGF-β1 表达增多[(194.816± 1.854)ng/l 比(112.716±7.484)ng/l，F=1160.819，P＜0.001]。2.与高糖组相比，SLX 作用后能明显上调HA[(196.119±2.260)pg/ml 比(172.074±3.223)pg/ml，F=120.845，P=0.013]和DCN[(3.227±0.067)ng/ml 比(2.859±0.175)ng/ml，F=15.413，P=0.008]的表达，明显下调IL-6[(67.927±2.467)ng/l 比(75.371± 2.386)ng/l，F=50.643，P=0.002]及TGF-β1[(232.778±31.029)ng/l 比(292.347±3.857)ng/l，F=21.458，P=0.002]的表达。结论HPMCs 存在HA 和DCN 的表达；高糖刺激可以反应性上调HPMCs 的HA 和DCN 表达，但其表达不足以保护腹膜，而补充舒洛地特后HA 和DCN 表达显著上调，具有修复受损糖萼的作用；高糖可以显著上调IL-6 及TGF-β1 的表达，舒洛地特可以抑制高糖刺激下HPMCs IL-6 和TGF-β1表达，从而抑制炎症和纤维化，保护腹膜功能。

关键词: 人腹膜间皮细胞, 糖萼, 舒洛地特, 透明质酸, 核心蛋白聚糖, IL-6, TGF-β1

Abstract: Objective To explore the effects of high glucose on the expression of glycocalyx including hyaluronic acid (HA) and decorin (DCN), inflammatory and fibrosis factors including IL-6 and TGF-β1 in human peritoneal mesothelial cells (HPMCs), and to observe the effects of sulodexide (SLX) on these changes in HPMCs. Methods HPMCs were randomly assigned into control group, high glucose group and high glucose+ SLX group. In the high glucose group, HPMCs were divided into different concentrations of glucose subgroups (HPMCs stimulated with 1.5%, 2.5% and 4.25% glucose for 48 hours) and different stimulation time subgroups (HPMCs stimulated with 2.5% glucose for 0, 6, 12, 24, 48 and 72 hours). In the high glucose+ SLX group, HPMCs were treated with 2.5% glucose and 50, 400, 800, 1200 μg/ml SLX for 48 hours. HA, DCN, IL-6 and TGF-β1 in the cultured media were assayed by ELISA. Results ①Compared to the control group, different concentrations of high glucose up-regulated the expressions of HA (172.074±3.223pg/ml vs. 147.000±1.667pg/ml, F=20.410, P=0.036), DCN (2.435±0.150ng/ml vs. 1.021±0.138ng/ml, F=219.555, P=0.000), IL-6 (67.981±1.655ng/l vs. 45.637±1.143ng/l, F=160.674, P=0.000) and TGF-β1 (204.691±2.829ng/l vs. 112.716±7.484ng/l, F=1692.284, P =0.000) in HPMCs. High concentrations of glucose for different hours also up- regulated the expression of HA (205.986 ± 2.746pg/ml vs. 147.000 ± 1.667pg/ml, F=1225.533, P=0.000), DCN (1.422±0.134ng/ml vs. 1.021± 0.138ng/ml, F=112.124, P=0.001), IL-6 (68.078±2.944ng/l vs.45.637±1.143ng/l, F=69.690, P=0.000), and TGF-β1 (194.816±1.854ng/l vs. 112.716±7.484ng/l, F=1160.819, P =0.000) in HPMCs. ② Compared to the high glucose group, treatment of SLX further increased the expression of HA (196.119±2.260pg/ml vs. 172.074±3.223pg/ml, F=120.845, P=0.013) and DCN (3.227±0.067ng/ml vs. 2.859±0.175ng/ml, F=15.413, P=0.008), but down-regulated the expression of IL-6 (67.927±2.467ng/l vs. 75.371 ± 2.386ng/l, F=50.643, P=0.002) and TGF- β1 (232.778 ± 31.029ng/l vs. 292.347 ± 3.857ng/l, F=21.458, P=0.002). Conclusion HPMCs expressed HA and DCN. Treatment of SLX further increased the high glucose induced up- regulation of HA and DCN, which may repair the injured activity of glycocalyx. SLX could reverse the high glucose induced up-regulation of IL-6 and TGF-β1 in HPMCs, which may protect peritoneal function through the inhibition of peritoneal inflammation and fibrosis.

Key words: HPMCs, Glycocalyx, SLX, HA, DCN, IL-6, TGF-β1