尹建勋, 帅怡, 王彦琴, 肖萍, 仲伟鉴. 活性氧在微囊藻毒素-LR诱导细胞凋亡效应中的作用探讨[J]. 环境与职业医学, 2011, 28(7): 398-401.
引用本文: 尹建勋, 帅怡, 王彦琴, 肖萍, 仲伟鉴. 活性氧在微囊藻毒素-LR诱导细胞凋亡效应中的作用探讨[J]. 环境与职业医学, 2011, 28(7): 398-401.
YIN Jian-xun , SHUAI Yi , WANG Yan-qin , XIAO Ping , ZHONG Wei-jian . Preliminary Study on the Role of ROS in the Apoptosis-inducing Effect of Microcystin-LR on Primary Cultured Rat Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2011, 28(7): 398-401.
Citation: YIN Jian-xun , SHUAI Yi , WANG Yan-qin , XIAO Ping , ZHONG Wei-jian . Preliminary Study on the Role of ROS in the Apoptosis-inducing Effect of Microcystin-LR on Primary Cultured Rat Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2011, 28(7): 398-401.

活性氧在微囊藻毒素-LR诱导细胞凋亡效应中的作用探讨

Preliminary Study on the Role of ROS in the Apoptosis-inducing Effect of Microcystin-LR on Primary Cultured Rat Hepatocytes

  • 摘要: 目的 研究微囊藻毒素-LR(MC-LR)对原代大鼠肝脏细胞的促凋亡效应以及探讨活性氧(ROS)在这一效应中的作用。

    方法 采用胶原酶灌注法获取原代大鼠肝脏细胞,用William's E培养液重悬后接种于铺被鼠尾胶原的孔板内,分为1个对照组和4个染毒组, 37℃、5% CO2条件下培养3 h后换液并染毒各梯度浓度的MC-LR,使各染毒组毒物终浓度分别为1& #215;10-5、1& #215;10-6、1& #215;10-7、1& #215;10-8 mol/L。染毒相应时间后,采用中性红比色实验检测细胞存活率,采用流式细胞术检测细胞凋亡和细胞内ROS含量。

    结果 中性红比色实验发现,至染毒期末观察到1& #215;10-5、1& #215;10-6 mol/L两个染毒组的肝脏细胞几乎全部死亡(>95%), 1& #215;10-7 mol/L染毒组部分肝脏细胞死亡(约30%);流式细胞术观察到MCLR染毒后早期(5 min)诱导细胞凋亡,同时观察到肝脏细胞内短时间(15 min)快速产生大量的ROS。

    结论 MC-LR诱导原代大鼠肝脏细胞快速产生大量的ROS,这可能是MC-LR诱导细胞凋亡并最终导致细胞死亡的毒性作用机制之一。

     

    Abstract: Objective To study the apoptosis-inducing effect of microcystin-LR (MC-LR) on primary cultured rat hepatocytes and the role of reactive oxygen species (ROS) in this effect.

    Methods Primary cultured rat hepatocytes were obtained through collagenase perfusion method, resuspended in William's E medium, and cultured in orifice plates coated with rattail collagen at 37℃, 5% CO2. They were divided into 1 control group and 4 dose groups, and treated with gradient concentrations of MC-LR. Neutral red colorimetric assay was used to detect the viability of hepatocytes, and flow cytometry to detect apoptosis as well as ROS level in hepatocytes.

    Results It was observed that the rat hepatocytes in two groups (MC-LR 1& #215;10-5, 1& #215;10-6 mol/L) almost totally died out, and in one group (MC-LR 1& #215;10-7 mol/L) died partly by neutral red colorimetric assay; and that MC-LR in duced rapid apoptosis and ROS formation by flow cytometry.

    Conclusion MC-LR induces rapid massive ROS in primary cultured rat hepatocytes, which may be one of the mechanisms that MC-LR induces apoptosis and death of the cells finally.

     

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