[吡咯烷二硫代氨基甲酸盐对急性百草枯中毒大鼠α-平滑肌肌动蛋白、整合素α5和纤连蛋白表达的影响]

[Effects of pyrrolidine dithiocarbamate on expressions of α-smooth muscle actin, integrin α5 and fibronectin in acute paraquat poisoned rats].

作者信息

Huang Min, Yang Hui-fang, Zhang Ping, Chang Xiu-li, Zhou Zhi-jun

机构信息

School of Public Health, Ningxia Medical University, Yinchuan 750004, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2013 May;31(5):341-6.

PMID:23803522

Abstract

OBJECTIVE

To observe the expressions of α-SMA, integrin α5 and fibronectin (Fn) in acute paraquat poisoned rats and the effect of PDTC. To investigate the mechanism of paraquat-induced pulmonary fibrosis.

METHODS

Sprague-Dawley rats were randomly divided into three experimental groups: Control group (6 rats), PQ group (36 rats) and PQ+PDTC group (36 rats). On the 1st, 3rd, the 7th, the 14th, the 28th and the 56th day after exposure, the protein expression of α smooth muscle actin (α-SMA) was evaluated by western blot. The mRNA levels of integrin α5 and fibronectin (Fn) were analyzed with real-time quantitative PCR (RT-PCR). Meanwhile, the lung pathological changes were observed and semi-quantified.

RESULTS

T With the time passing, the expression of α-SMA in PQ group increased gradually compared with control group (P < 0.05 or P < 0.01). The increasing extent was gently on the 3 rd, the 7 th day. While increasing extent was rapidly from the 28 th to the 56 th day. RT-PCR showed PQ significantly increased Fn mRNA level on all time points and increased integrin α5 mRNA level from the 7 rd to 56 th day compared with control group (P < 0.05 or P < 0.01). PDTC treatment significantly deceased α-SMA, Fn, and integrin α5 levels compared with PQ group in corresponding time points (P < 0.05 or P < 0.01) Noteworthy, in PQ+PDTC group, the occurrence of pathological changes were drastically attenuated and pathologic score significantly decreased (P < 0.05 or P < 0.01).

CONCLUSIONS

α-SMA, integrin α5 and fibronectin could play an important role in the development of pulmonary fibrosis caused by paraquat poisoning. PDTC, asa strong NF-κB inhibitor, may inhibit NF-κB activity and further significantly decreased expressions of α-SMA, integrin α5 and fibronectin which were important part of ECM, leading to drastically attenuated pulmonary fibrosis. However, the mechanisms of PDTC intervention still remains to be explored.

摘要

目的

观察急性百草枯中毒大鼠α -平滑肌肌动蛋白（α - SMA）、整合素α5和纤连蛋白（Fn）的表达情况以及吡咯烷二硫代氨基甲酸盐（PDTC）的作用。探讨百草枯致肺纤维化的机制。

方法

将Sprague - Dawley大鼠随机分为三个实验组：对照组（6只大鼠）、百草枯组（36只大鼠）和百草枯 + PDTC组（36只大鼠）。在染毒后第1天、第3天、第7天、第14天、第28天和第56天，采用蛋白质免疫印迹法检测α - 平滑肌肌动蛋白（α - SMA）的蛋白表达。用实时定量聚合酶链反应（RT - PCR）分析整合素α5和纤连蛋白（Fn）的mRNA水平。同时，观察并对肺组织病理变化进行半定量分析。

结果

随着时间推移，百草枯组α - SMA的表达与对照组相比逐渐增加（P < 0.05或P < 0.01）。在第3天和第7天增加程度较平缓，而从第28天到第56天增加迅速。RT - PCR结果显示，百草枯组在各个时间点均显著增加Fn的mRNA水平，且从第7天到第56天整合素α5的mRNA水平也增加，与对照组相比差异有统计学意义（P < 0.05或P < 0.01）。与百草枯组相应时间点相比，PDTC治疗显著降低了α - SMA、Fn和整合素α5的水平（P < 0.05或P < 0.01）。值得注意的是，在百草枯 + PDTC组，病理变化的发生明显减轻，病理评分显著降低（P < 0.05或P < 0.01）。

结论

α - SMA、整合素α5和纤连蛋白在百草枯中毒所致肺纤维化的发生发展中可能起重要作用。PDTC作为一种强效的核因子κB（NF - κB）抑制剂，可能抑制NF - κB活性，进而显著降低作为细胞外基质重要组成部分的α - SMA、整合素α5和纤连蛋白的表达，导致肺纤维化明显减轻。然而，PDTC干预的机制仍有待进一步探索。