[姜黄素对大鼠肺成纤维细胞凋亡的影响]

[Effects of curcumin on apoptosis of pulmonary fibroblasts from rats].

作者信息

Gao Wei, Zhang De-Ping, Chen Bi

机构信息

Department of Respiratory Medcine, Affiliated Drum Tower Hospital of Nanjing University, Nanjing 210008, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2009 Jan;32(1):37-41.

PMID:19484960

Abstract

OBJECTIVE

To investigate the effect of curcumin on apoptosis of pulmonary fibroblasts from rats and the molecular mechanisms.

METHODS

Twenty 6 week old male SD rats were randomly divided into 2 groups with 10 rats in each group: the normal control group (NLF) and the model group (MLF). The pulmonary fibrosis models were made by intratracheally instillation of bleomycin. The rats were sacrificed on the 28 day of experiment, and the pathologic changes of the lung tissues were investigated. Fibroblasts were derived from lung tissues and were cultured in vitro. Then the cells were treated with curcumin at different concentrations (5, 10, 20 and 40 micromol/L). TUNEL staining was used to detect apoptosis. The Caspase-3 and Caspase-8 expression of each group were determined by flow cytometry (FCM). The expression changes of apoptosis-regulating proteins of Caspase-9 in cells were examined by immunohistochemical staining.

RESULTS

In the MLF group treated with 4 different concentrations of curcumin for 12 h, apoptotic morphological changes were observed in the cells. Apoptotic index (AI) was (30.17 +/- 0.53)%, (37.20 +/- 0.25)%, (45.12 +/- 0.31)%, (51.90 +/- 0.27)% respectively, which were significantly higher than that of the control group without curcumin (4.59 +/- 0.19)%. The expression of Caspase-3 was (53.1 +/- 0.7)%, (60.5 +/- 0.6)%, (69.1 +/- 0.9)%, (74.9 +/- 0.7)%, which were significantly higher than that of the control group (5.5 +/- 0.8)%. The expression of Caspase-8 were (37.2 +/- 0.5)%, (46.8 +/- 0.4)%, (56.5 +/- 0.3)%, (67.3 +/- 0.3)%, which were significantly higher than that of the control group (4.6 +/- 0.6)%. The expression of Caspase-9 were (39.38 +/- 0.88)%, (47.51 +/- 0.24)%, (57.75 +/- 0.63)%, (68.36 +/- 0.29)%, which were significantly higher than that of the control group (4.92 +/- 0.34)%. Curcumin did not induce apoptosis of NLF, the expressions of Caspase-3, Caspase-8, Caspase-9 being not statistically different from that of the control group without curcumin.

CONCLUSIONS

Curcumin can induce apoptosis of pulmonary fibroblasts in rats with bleomycin-induced pulmonary fibrosis and the mechanism may be related to the activation of Caspase-3, Caspase-8, and Caspase-9.

摘要

目的

探讨姜黄素对大鼠肺成纤维细胞凋亡的影响及其分子机制。

方法

将20只6周龄雄性SD大鼠随机分为2组，每组10只：正常对照组（NLF）和模型组（MLF）。采用气管内滴注博来霉素制备肺纤维化模型。实验第28天处死大鼠，观察肺组织病理变化。从肺组织中分离出成纤维细胞并进行体外培养。然后用不同浓度（5、10、20和40微摩尔/升）的姜黄素处理细胞。采用TUNEL染色检测细胞凋亡。通过流式细胞术（FCM）测定每组Caspase-3和Caspase-8的表达。采用免疫组织化学染色检测细胞中凋亡调节蛋白Caspase-9的表达变化。

结果

用4种不同浓度姜黄素处理MLF组细胞12小时后，观察到细胞出现凋亡形态学变化。凋亡指数（AI）分别为（30.17±0.53）%、（37.20±0.25）%、（45.12±0.31）%、（51.90±0.27）%，均显著高于未用姜黄素处理的对照组（4.59±0.19）%。Caspase-3的表达分别为（53.1±0.7）%、（60.5±0.6）%、（69.1±0.9）%、（74.9±0.7）%，均显著高于对照组（5.5±0.8）%。Caspase-8的表达分别为（37.2±0.5）%、（46.8±0.4）%、（56.5±0.3）%、（67.3±0.3）%，均显著高于对照组（4.6±0.6）%。Caspase-9的表达分别为（39.38±0.88）%、（47.51±0.24）%、（57.75±0.63）%、（68.36±0.29）%，均显著高于对照组（4.92±0.34）%。姜黄素未诱导NLF凋亡，Caspase-3、Caspase-8、Caspase-9的表达与未用姜黄素处理的对照组相比无统计学差异。