中国血液净化

›› 2008, Vol. 7 ›› Issue (6): 325-328.

• 基础研究 • 上一篇    下一篇

去甲斑蝥素对白蛋白刺激肾小管上皮细胞增殖及上调纤维连接蛋白表达的影响

叶 琨 刘伏友 李 瑛 彭佑铭 段绍斌 刘 虹   

  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-06-12 发布日期:2008-06-12
  • 通讯作者: 刘伏友

Effects of norcantharidin on proliferation and fibronectin expression in HK-2 cells induced by albumin

YE Kun, LIU Fu-you, LI Ying, PENG You-ming, DUAN Shao-wu, LIU Hong   

  • Received:1900-01-01 Revised:1900-01-01 Online:2008-06-12 Published:2008-06-12

RichHTML

0

PDF

238

摘要： 【摘要】目的 观察去甲斑蝥素(NCTD)对白蛋白刺激人近端肾小管上皮细胞株(HK-2)增殖及上调FN表达的影响，初步探讨NCTD延缓肾间质纤维化的体外作用机制。 方法 通过细胞毒性实验筛选出对HK-2细胞无明显损伤的NCTD适合浓度。将HK-2细胞随机分为5组：对照组(1640培基组)，白蛋白组(1640＋白蛋白5g/L)，各浓度NCTD组(1640＋白蛋白5g/L＋NCTD 0.5，1，2.5mg/L)。在各组HK-2细胞中加入相应干预液孵育24h后，采用细胞增殖法(MTT法)检测NCTD对白蛋白刺激HK-2细胞增殖的影响；酶联免疫吸附试验 (ELISA) 测定各组细胞上清中FN的蛋白含量；RT-PCR法半定量分析HK-2细胞中FN mRNA的表达。 结果 ①比较对照组及NCTD (0.5，1.0，2.5 mg/L) 各组的乳酸脱氢酶(LDH)浓度，各组间的差异无统计学意义(P＞0.05)；②白蛋白组MTT值(0.438±0.073)明显高于对照组(0.330±0.060)(P＜0.05)，而NCTD(2.5 mg/L) 组MTT值(0.327±0.080) 较白蛋白组降低 (P＜0.05)；③与白蛋白组比较，NCTD (2.5mg/L)组HK-2细胞上清中FN蛋白含量降低，差异有统计学意义(P＜0.01); NCTD(1mg/L和2.5mg/L)组细胞上清中FN mRNA的表达较白蛋白组明显下调，差异有统计学意义(P＜0.01)。 结论 白蛋白(5g/L)诱导HK-2细胞增殖及FN过表达，一定浓度去甲斑蝥素可抑制白蛋白对HK-2细胞的活化作用。

关键词: 斑蝥素, 人近端肾小管上皮细胞, MTT法, 纤维连接蛋白

Abstract: Objective To observe the effects of norcantharidin (NCTD) on proliferation and fibronectin (FN) expression in human renal proximal tubular epithelial cell line (HK-2) induced by albumin, and to investigate the mechanism of NCTD in the prevention of renal interstitial fibrosis. Methods Appropriate NCTD concentration which did not significantly injure HK-2 cells was determined by cytotoxicity test. HK-2 cells were divided into 5 groups: control group (1640 group), albumin group (1640 + albumin 5g/L), and NCTD 0.5 mg/L, 1mg/L and 2.5mg/L groups (1640 + albumin 5g/L + NCTD 0.5, 1 or 2.5mg/L). HK-2 cells were incubated with respective concentrations of NCTD and albumin for 24 hours. Proliferation of HK-2 cells was determined by MTT, and FN protein in culture media by enzyme-linked immunosorbent assay (ELISA). FN mRNA in HK-2 cells was analyzed by RT-PCR. Results ①Cytotoxicity test showed that there were no significant differences in lactate dehydrogenase (LDH) concentration among NCTD 0.5 mg/L，1.0 mg/L and 2.5 mg/L groups as well as control group (P>0.05); ② MTT score was higher in albumin group (0.438+0.073) than in 1640 group (0.330+0.060) (P<0.05), but MTT score decreased to 0.327+0.080 in NCTD 2.5mg/L group (P<0.05, as compared with that of albumin group); ③FN protein was down-regulated in NCTD 2.5mg/L group (P<0.01, as compared with that of albumin group), and FN mRNA significantly decreased in NCTD 1mg/L and 2.5mg/L groups (P<0.01, as compared with that of albumin group). Conclusion NCTD inhibits the albumin-induced cell proliferation and FN over-expression in HK-2 cells.

中图分类号: