中国医学科学院学报

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中国医学科学院学报

中国医学科学院学报 ›› 2007, Vol. 29 ›› Issue (3): 364-369.

• 论著 • 上一篇    下一篇

Janus激酶抑制剂对高糖诱导的肾小管上皮细胞转分化的作用

张勉之1,张敏英2,赵 松3,段建召3,张艳秋1,左春霞1,程项阳1,段惠军3   

  1. 1天津市公安医院肾内科,天津 300040
    2天津医科大学流行病学教研室,天津 300070
    3河北医科大学病理教研室,石家庄 050017
  • 收稿日期:2007-01-05 修回日期:1900-01-01 出版日期:2007-06-30 发布日期:2007-06-30
  • 通讯作者: 张勉之

Effect of JAK/ STAT Pathway Activation on High Glucose-induced
Transdifferentiation in Renal Proximal Tubular Epithelial Cells

ZHANG Mian-zhi1,ZHANG Min-ying2,ZHAO Song3,DUAN Jian-zhao3,ZHANG Yan-qiu1, ZUO Chun-xia1,CHENG Xiang-yang1,DUAN Hui-jun3   

  1. 1Department of Nephrology,Tianjin Gongan Hospital,Tianjin 300040,China
    2Department of Epidemiology,Tianjin Medical University,Tianjin 300070,China
    3 Department of Pathology, Hebei Medical University,Shijiazhang 050017,China
  • Received:2007-01-05 Revised:1900-01-01 Online:2007-06-30 Published:2007-06-30
  • Contact: ZHANG Mian-zhi

摘要: 摘要:目的 评估Janus激酶抑制剂AG490对高糖诱导肾小管上皮细胞转分化及转化生长因子β1(TGF-β1)表达的影响。方法 体外培养人肾近曲小管上皮细胞株(HKC),分别给予高糖和高糖+AG490干预, Western blot检测平滑肌肌动蛋白(α-SMA)、E-钙粘素(E-Cadherin)及信号蛋白STAT1、STAT3、磷酸化STAT1 (p-STAT1) 和p-STAT3的表达,酶联免疫吸附法测定细胞上清液中TGF-β1和Ⅰ型胶原的分泌,逆转录聚合酶链反应检测TGF-β1mRNA表达。结果 与低糖对照组比较,高糖培养的HKC中α-SMA 、p-STAT1 和p-STAT3 表达明显上调,E-Cadherin表达明显下调,TGF-β1 mRNA 表达增加,细胞培养上清液中TGF-β1和Ⅰ型胶原分泌增加。AG490明显下调p-STAT1和p-STAT3 表达的同时,明显抑制高糖刺激HKC中α-SMA表达的升高,减轻E-Cadherin表达下降程度,降低TGF-β1 mRNA 表达及TGF-β1和Ⅰ型胶原的分泌。结论 JAK参与了高糖诱导的HKC转分化,并刺激TGF-β1和细胞外基质分泌,JAK抑制剂AG490能有效拮抗其作用。

关键词: 人肾近曲小管上皮细胞株, 细胞外基质, 转化生长因子β1

Abstract: ABSTRACT:Objective To evaluate the effect of JAK/STAT signaling pathway activation on the transdifferentiation and secretion of transforming growth factor-β1 ( TGF-β1 ) induced by high glucose in renal proximal tubular epithelial cells. Methods Human kidney cells(HKC) were cultured and then divided into four groups: low glucose (LG) group, high glucose (HG) group, high mannitol ( LG+M) group, and HG+AG490 group. Immunoprecipitation and Western blot analysis were used to determine the expression of tryosine phosphorylated Janus kinase 2 (p-JAK2) . The protein expressions of STAT1, STAT3, p-STAT1, and p-STAT3 and the expressions of α-SMA and E-Cadherin were observed by Western blot. The contents of TGF-β1, fibronectin and type I collagen in the supernatants of the cultured HKC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-β1 mRNA was measured by reverse transcription and polymerase chain reaction (RT-PCR). Results Compared with LG group, the expressions of JAK2, p-STAT1, p-STAT3, and TGF-β1 mRNA were significantly increased in HG group from 6 to 72 hours. Meanwhile, the contents of TGF-β1 and collagen I in the supernatants and the expression of α-SMA increased and the expression of E-Cadherin decreased. The expressions of JAK2, p-STAT1, p-STAT3, and TGF-β1mRNA as well as the levels of TGF-β1 and collagen I in the supernatant s in HG+AG490 group were significantly lower than in the HG group. The expressions of α-SMA and E-Cadherin were also decreased in HG+AG490 group. Conclusion Activation of JAK/STAT signaling pathway may be involved in the high glucose- induced transdifferentiation and overproduction of TGF-β1 and ECM proteins in HKCs.

Key words: duman kidney cell, transdifferentiation, transforminc crowtd factor-β1