天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (5): 465-469.doi: 10.11958/j.issn.0253-9896.2015.05.005

• 细胞与分子生物学 • 上一篇    下一篇

五味子乙素对HK-2细胞缺氧损伤的保护作用

卢爱龙 1， 谭小月 2， 张勉之 3△， 吴银娜 1   

  1. 1天津医科大学 （邮编 300070）； 2南开大学医学院； 3天津市公安医院
  • 收稿日期:2015-03-05 修回日期:2015-03-19 出版日期:2015-05-15 发布日期:2015-05-25
  • 通讯作者: 张勉之 E-mail： zhangmianzhi@vip.sina.com E-mail:834633135@qq.com
  • 作者简介:卢爱龙 （1988）， 男， 硕士研究生在读， 主要从事肾缺血再灌注损伤研究
  • 基金资助:
    天津市应用基础与前沿技术研究计划 （14JCYBJC28200）

Protective effects of schisandrin B on hypoxia injury of HK-2 cells

LU Ailong1, TAN Xiaoyue2, ZHANG Mianzhi3△, WU Yinna1#br# #br#   

  1. 1 Tianjin Medical University, Tianjin 300070, China; 2 Medical School of Nankai University; 3 Tianjin Gongan Hospital
  • Received:2015-03-05 Revised:2015-03-19 Published:2015-05-15 Online:2015-05-25
  • Contact: ZHANG Mianzhi,E-mail： zhangmianzhi@vip.sina.com E-mail:834633135@qq.com

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摘要： 卢爱龙 1， 谭小月 2， 张勉之 3△， 吴银娜摘要： 目的 探讨五味子乙素 （Sch B） 对氯化钴 （CoCl2） 诱导的人类近端肾小管上皮 （HK-2） 细胞缺氧损伤的保护作用及其可能机制。方法 取离体培养 HK-2 细胞， 随机分为 4 组。对照 （C） 组： 细胞未经任何处理。CoCl2组 （化学乏氧组）： 加入 600 μmol/L 的 CoCl2 处理 24 h。Sch B 预保护（CoCl2+ Sch B）组： 分别加入终浓度为 1 μmol/L 和 10 μmol/L Sch B 预处理 2 h 后， 其余操作同 CoCl2 组。Sch B 组： 分别加入终浓度 1 μmol/L 和 10 μmol/L Sch B 处理 2 h。CCK-8 试剂盒检测各组细胞活性； AnnexinV-FITC/PI 双标记流式细胞仪检测各组细胞凋亡率； Western Blot 检测各组缺氧诱导因子-1α（HIF-1α）蛋白表达； RT-PCR 检测各组 HIF-1α和诱导型一氧化氮合酶（iNOS） mRNA 表达。结果 与对照组相比， CoCl2组细胞活性明显降低， 细胞凋亡率、 HIF-1α蛋白表达量和 iNOS mRNA 表达量显著增加， HIF-1α mRNA 表达量差异无统计学意义； Sch B 预保护组较 CoCl2组细胞活性显著增加， 细胞凋亡率、 HIF-1α蛋白表达量、 HIF-1α及 iNOS mRNA 表达量均显著减少； Sch B 组与对照组细胞活性、 细胞凋亡率差异无统计学意义， Sch B 组几乎不表达 HIF-1α蛋白。结论 Sch B 可能通过抑制 HIF-1α蛋白和 iNOS mRNA 的表达减少 HK-2 细胞的凋亡， 从而对 HK-2 细胞缺氧损伤起保护作用。

关键词: 五味子乙素, HK-2细胞, 氯化钴, 缺氧损伤, 缺氧诱导蛋白-1α(HIF-1α), 诱导型一氧化氮合酶(iNOS)

Abstract: Abstract： Objective To explore the protective effects of schisandrin B (Sch B) on hypoxia injury induced by cobal⁃ tous chloride (CoCl2) in human proximal renal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Meth⁃ ods HK-2 cells were randomly assigned to four groups: control group (Con, cells were untreated), CoCl2 group (CoCl2, cells were treated with 600 μmol/L CoCl2 for 24 h), Sch B pretreat group (CoCl2+Sch B, cells were pretreated with 1 μmol/L and 10 μmol/L Sch B for 2 h) and Sch B group (Sch B, cells were treated with 1 μmol/L and 10 μmol/L Sch B for 2 h). CCK-8 kit was used to detect the cell viability of four groups. Flow cytometry was used to detect the apoptotic rate of four groups. The protein expression of hypoxia-inducible factor 1α (HIF-1α) was assessed by Western blot assay. The expressions of HIF-1α and inducible nitric oxide synthase (iNOS) mRNA were determined by RT-PCR. Results Compared with the control group, after treated with 600 μmol/L CoCl2, the cell viability was decreased, and the apoptosis was increased, the expressions of HIF-1α and iNOS mRNA were up-regulated in HK-2 cells. There was no significant difference in the expression of HIF- 1α mRNA between control group and CoCl2 group. Compared with the CoCl2 group, after pretreated with 1 μmol/L and 10 μmol/L Sch B, the cell viability was increased and the apoptosis was decreased, the expressions of HIF-1α and iNOS were down-regulated in HK-2 cells. There were no significant differences in the cell viability and apoptotic rate between control group and Sch B group. Conclusion Pretreatment with Sch B can reduce the apoptosis of HK-2 cells by inhibiting the ex⁃ pression of HIF-1α and iNOS mRNA, which shows protective effects on hypoxia injury.

Key words: schisandrin B, HK-2 cell, CoCl2, hypoxia injury, HIF-1α, iNOS