硫酸锌诱导金属硫蛋白减轻再灌注肺细胞凋亡的实验研究

中国临床药理学与治疗学 ›› 2010, Vol. 15 ›› Issue (10): 1110-1115.

硫酸锌诱导金属硫蛋白减轻再灌注肺细胞凋亡的实验研究

石璐^1,2, 贾旭广^1,2, 邱晓晓¹, 钱小英³, 汪洋¹, 许冬武¹, 王万铁¹

¹温州医学院病理生理教研室,温州 325035,浙江;
²桐乡市卫生学校,桐乡 314500,浙江;
³温州市第三人民医院呼吸内科,温州 325000,浙江

收稿日期:2010-03-30 修回日期:2010-06-29 发布日期:2020-09-16
通讯作者: 王万铁,男,教授,硕士生导师,研究方向:缺血/再灌注损伤。E-mail: wzwwt@tom.com
作者简介:石璐,女,硕士,研究方向:缺血/再灌注损伤。E-mail: ivy-1616@hotmail.com
基金资助:
温州市科技计划项目(Y20060083);温州市医药卫生科研项目(2009A004)

Effects of metallothionein induced by ZnSO₄ on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats

SHI Lu^1,2, JIA Xu-guang^1,2, QIU Xiao-xiao¹, QIAN Xiao-ying², WANG Yang¹, XU Dong-wu², WANG Wan-tie¹

¹Department of Pathophysiology, Wenzhou Medical College, Wenzhou 325035, Zhejiang, China;
²Health School of Tongxiang, Tongxiang 314500, Zhejiang, China;
³Department of Respiratory, the 3rd People's Hospital of Wenzhou 325035, Zhejiang, China

Received:2010-03-30 Revised:2010-06-29 Published:2020-09-16

摘要/Abstract

摘要： 目的: 观察硫酸锌诱导金属硫蛋白对再灌注肺细胞凋亡的影响。方法: 健康雄性SD大鼠24只,随机分为对照组(C组)、模型组(I/R组)和研究组 (M组)。对比观察各组肺组织中金属硫蛋白的含量,血清中丙二醛(MDA)、超氧化物歧化酶(SOD)、髓过氧化物酶(MPO)活力及含量变化,原位缺口末端标记法(TUNEL)检测肺组织细胞凋亡情况,免疫组化及RT-PCR法检测肺组织中Bax、Bcl-2蛋白和基因的表达,透射电镜观察肺组织超微结构的改变。结果: I/R组与C组相比,肺组织中金属硫蛋白的含量显著下降,MDA含量、MPO活力明显升高,SOD活力明显下降(P<0.05 或P<0.01),肺组织原位细胞凋亡检测显示I/R组凋亡指数(AI)39.03±3.46 显著高于C组 2.88±0.34,Bcl-2/Bax 比值在蛋白和基因水平明显降低(P<0.05 或P<0.01),肺组织超微结构发生异常改变;金属硫蛋白组与I/R组相比肺组织中金属硫蛋白的含量显著升高(P<0.01),MDA含量、MPO活力明显下降,SOD活力明显升高(P<0.05 或P<0.01),AI为 15.50±1.02 显著低于I/R组,并能明显升高Bcl-2/Bax 比值及改善肺组织超微结构。结论: 金属硫蛋白通过减轻脂质过氧化反应及中性粒细胞聚集,降低Bax/Bcl-2 比值,使肺组织细胞凋亡减少,从而有效地减轻肺缺血/再灌注损伤。

关键词: 肺, 缺血/再灌注损伤, 金属硫蛋白, 硫酸锌, 凋亡

Abstract: AIM: To investigate the effects of metallothionein induced by ZnSO₄ on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats. METHODS: Adult male Sprague-Dawley rats were randomly divided into 3 groups based upon the intervention (n=8): control group (C), LIR group (I/R), LIR+ ZnSO₄ group (M). At the end of the experiment, the content of metallothionein was tested in lung tissue; blood specimens drawn from the arteria carotis were tested for the content of Malondialdehyde (MDA), the activity of Superoxide dismutase (SOD) and Myeloperoxidase (MPO); the pneumocyte apoptosis index (AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end abeling (TUNEL); the expression of Bcl-2, Bax protein in lung tissue were accessed by quantitative immunohistochemistry (IHC) and Bcl-2, Bax mRNA by RT-PCR; ultrastructural changes of lung tissue were ob- served by transmission electron microscope. RESULTS: Metallothionein induced by ZnSO₄ could significantly attenuate the MDA level, MPO activity and improve SOD activity in blood serum which was comparable to I/R and significantly reduced the number of TUNEL-positive cells vs. the I/R group, expressed as AI (% total nuclei) from 39.03±3.46 to 15.50±1.02 (P<0.01). The protein and mRNA expression of Bcl-2, Bax showed that metallothionein significantly attenuated the ischemia/ reperfusion-upregulated expression of Bax protein but improved the expression of Bcl-2 that improved the Bcl-2/ Bax ratio(P<0.05 or P <0.01). There were abnormal changes of the ultrastructure in I/R, and markedly reversed in MT group. CONCLUSION: Metallothionein induced by ZnSO₄ may attenuate pneumocyte apoptosis in LIRI by up-regulating expression of Bcl-2/ Bax ratio and by inhibiting oxidant generation, neutrophils filtration.

Key words: Lung, Ischemia/reperfusion injury, Metallothionein, ZnSO₄, Apoptosis

中图分类号:

R363

石璐, 贾旭广, 邱晓晓, 钱小英, 汪洋, 许冬武, 王万铁. 硫酸锌诱导金属硫蛋白减轻再灌注肺细胞凋亡的实验研究[J]. 中国临床药理学与治疗学, 2010, 15(10): 1110-1115.

SHI Lu, JIA Xu-guang, QIU Xiao-xiao, QIAN Xiao-ying, WANG Yang, XU Dong-wu, WANG Wan-tie. Effects of metallothionein induced by ZnSO₄ on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(10): 1110-1115.

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