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[N-乙酰半胱氨酸对六价铬诱导的L-02肝细胞凋亡的拮抗作用(有无半胱天冬酶抑制剂)]

[Antagonistic effect of N-acetylcysteine on apoptosis of L-02 hepatocyte induced by Cr(VI) with or without caspase inhibitor].

作者信息

Chen Jing, Zhong Caigao, Zeng Ming, Liu Xinmin, Deng Yuanyuan, Xiao Fang

机构信息

Department of Health Toxicology, School of Public Health, Central South University, Changsha 410078, China.

出版信息

Wei Sheng Yan Jiu. 2010 Nov;39(6):678-81.

Abstract

OBJECTIVE

To explore the antagonistic effect of N-acetylcysteine (NAC) on hexevalent chromium (Cr(VI))-induced apoptosis in L-02 hepatocytes with or without caspase inhibitors.

METHODS

L-02 hepatocytes were randomly divided into a control group, and Cr( VI), Z-VAD-fmk + Cr(VI), NAC + Cr(VI), Z-VAD-fmk + NAC + Cr (VI) four treatment groups, in which L-02 hepatocytes were cultured with Cr (VI) at the dose of 20 micromol/L for 6h. The rates of apoptosis in all groups were detected by flow cytometry (FC) after staining with propidium iodide (PI). The changes of mitochondrial membrane potential (deltapsim) and permeability transition pore (PTP) were determined by fluorescent spectrometer. The DNA damages in hepatocytes were observed by the single cell gel electrophoresis (SCGE).

RESULTS

Cr(VI) significantly induced apoptosis of L-02 hepatocytes at the dose of 20 micromol/L for 6 hours (P < 0.05). However, NAC significantly decreased the rates of apoptosis of L-02 hepatocytes and alleviated the damages to mitochondria and DNA caused by Cr(VI) in L-02 hepatocytes with or without caspase (P < 0.05). However, in comparition with the non caspase-inhibited group, the protective effects of NAC decreased in the caspase-inhibited group (P < 0.05).

CONCLUSION

NAC could protect the apoptosis of L-02 hepatocyte induced with Cr(VI) with or without caspase inhibitor, and caspase could not play a decisive role in this process.

摘要

目的

探讨N-乙酰半胱氨酸(NAC)对六价铬(Cr(VI))诱导的L-02肝细胞凋亡的拮抗作用,以及有无半胱天冬酶抑制剂时的情况。

方法

将L-02肝细胞随机分为对照组、Cr(VI)组、Z-VAD-fmk + Cr(VI)组、NAC + Cr(VI)组、Z-VAD-fmk + NAC + Cr(VI)组,其中用20微摩尔/升的Cr(VI)培养L-02肝细胞6小时。用碘化丙啶(PI)染色后,通过流式细胞术(FC)检测所有组的细胞凋亡率。用荧光光谱仪测定线粒体膜电位(Δψm)和通透性转换孔(PTP)的变化。通过单细胞凝胶电泳(SCGE)观察肝细胞中的DNA损伤。

结果

20微摩尔/升的Cr(VI)作用6小时可显著诱导L-02肝细胞凋亡(P < 0.05)。然而,NAC可显著降低L-02肝细胞的凋亡率,并减轻Cr(VI)对L-02肝细胞线粒体和DNA的损伤,无论有无半胱天冬酶(P < 0.05)。然而,与未用半胱天冬酶抑制剂的组相比,NAC在半胱天冬酶抑制剂组中的保护作用降低(P < 0.05)。

结论

NAC可保护Cr(VI)诱导的L-02肝细胞凋亡,无论有无半胱天冬酶抑制剂,且半胱天冬酶在该过程中不起决定性作用。

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