[1]王梓华,黄云剑.马兜铃酸对人肾小管上皮细胞损伤的体外实验研究[J].第三军医大学学报,2009,31(23):2302-2305.
 WANG Zi-hua,HUANG Yun-jian.Mechanism of aristolochic acid induced injury in human renal tubular epithelial cells in vitro[J].J Third Mil Med Univ,2009,31(23):2302-2305.
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马兜铃酸对人肾小管上皮细胞损伤的体外实验研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第23期
页码:
2302-2305
栏目:
论著
出版日期:
2009-12-15

文章信息/Info

Title:
Mechanism of aristolochic acid induced injury in human renal tubular epithelial cells in vitro
作者:
王梓华黄云剑
第三军医大学新桥医院肾内科,全军肾脏病中心,重庆市肾病研究所
Author(s):
WANG Zi-hua HUANG Yun-jian
Department of Nephrology, Renal Disease Research Center, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
关键词:
马兜铃酸肾小管上皮细胞转分化凋亡
Keywords:
aristolochic acid human renal tubular epithelial cells transdifferention apoptosis
分类号:
R392.12
文献标志码:
A
摘要:
目的    研究马兜铃酸(aristolochic acid,AA)对人近端肾小管上皮细胞(HK-2细胞)的损伤及可能机制。    方法    将细胞分为4组:正常对照组与AA30、60、120 μmol/L组(n=6),分别作用于HK-2细胞培养48 h后,倒置相差显微镜观察细胞形态,CCK8试剂盒(Cell Counting Kit-8)检测增殖,流式细胞仪检测凋亡,Western blot分析激活型Caspase-3的表达,全自动生化检测仪测定上清液中乳酸脱氢酶(LDH)和β-N-乙酰氨基葡萄糖苷酶(NAG酶)的含量,激光共聚焦扫描荧光显微镜(laser scanning confocal microscope,LSCM)观察α-平滑肌肌动蛋白(smooth muscle actin,α-SMA)和E-钙黏连蛋白(E-cadherin)的表达, ELISA测定上清液中转化生长因子-β1(transforming growth factor-β1,TGF-β1)和Ⅲ型胶原的分泌。    结果    HK-2分别在30、60、120 μmol/L AA作用48 h后,出现增殖抑制,凋亡增加,激活型Caspase-3表达增多,LDH和NAG酶升高,均呈剂量依赖性。60 μmol/L浓度的AA还表现Ecadherin表达减弱,α-SMA表达增强,TGF-β1和Ⅲ型胶原分泌明显增加(P<0.05)。    结论    AA可引起HK-2细胞明显的增殖抑制、凋亡和上皮-间充质转分化呈一定的浓度依赖性。
Abstract:
Objective    To investigate the possible injury mechanism of human kidney proximal tubular epithelial cell-2 (HK-2) induced by aristolochic acid (AA).     Methods     Cultured HK-2 cells were divided into 4 groups: normal control, treated by AA at the concentration of 30, 60 and 120 μmol/L for 48 h respectively. The morphological changes were observed by inverted phase contract microscopy. The cell viability was measured by the Cell Counting Kit-8 (CCK8) assay. Apoptotic cells were identified by flow cytometry. Expression of active Caspase-3 was measured by Western blot analysis. Automatic biochemical analyzer was used to detect the contents of LDH and β-N-Acetylglucosaminidase (NAG) in the supernatant. The expression of E-cadherin and a-SMA was detected with laser scanning confocal microscope (LSCM). Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of TGF-β1 and collagen Ⅲ in the supernatant quantitatively.     Results    AA inhibited HK-2 cells proliferation, induced cell apoptosis and activated Caspase-3 expression, and increased the LDH and NAG levels. All of these were in a concentration-dependent manner. AA at the concentration of 60 μmol/L inhibited E-cadherin expression, increased α-SMA expression and TGF-β1 and collagen Ⅲ secretion.     Conclusion    AA inhibits cell proliferation, induces apoptosis and epithelial-mesenchymal transition (EMT) in HK-2 cells. AA at relatively low concentration (≤60 μmol/L) mainly induces EMT in HK-2 cells, while, that at high concentration (≥120 μmol/L) causes apoptosis and cytotoxicity.

参考文献/References:

王梓华,黄云剑.马兜铃酸对人肾小管上皮细胞损伤的体外实验研究[J]. 第三军医大学学报,2009,31(23):2302-2305.

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更新日期/Last Update: 2009-11-23