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[抑制矽肺纤维化的实验研究]

[The experimental study of suppressing silicosis fibrosis].

作者信息

Weng Ze-ping, Zhang Ji-jun, Liu Wei-wei, Chen Juan, Liu Yi-min, Yu Wei, Tang Li-juan, Chen Jia-yu, Fang Mao, Zhang Cheng, Ye Geng-xin, Chen Ling-zhen, Zhong Xue-yun

机构信息

Department of Pathology, Medical College of Jinan University, Guangzhou 510632, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2011 Oct;29(10):740-5.

Abstract

OBJECTIVE

To compare the difference of effects on SiO(2)-induced alveolitis and early fibrosis between bone marrow-derived mesenchymal-like stem cells (BM-MSCs) and BM-MSCs transfected by pcDNA3.1-HGF and to explore the mechanism of this effects.

METHODS

The Primary BM-MSCs from Wistar male young rats were cultured and labeled by 4, 6-diamidino-2-phenylindole (DAPI). Fifty Wistar rats were randomly divided into 3 groups:model group (10 rats),which was administered with SiO(2) by the trache, the next day,injected PBS via the tail vein; BM-MSCs group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs via the tail vein; pcDNA3.1-HGF plus BM-MSC group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs transfected by pcDNA3.1-HGF via the tail vein. On the 14th and 28th days after treatment, half of the animals were sacrificed, respectively, and the lungs were harvested for frozen section to observe the cell marked by DAPI. HE staining under a fluorescent microscope, and to observe the pulmonary alveolitis and fibrosis by HE and Masson staining under a light microscope. Western blot assay was used to detect the expression of HGF in rat lungs. The expression levels of tumor necrosis factor-α (TNF-α) in pulmonary tissues were analyzed quantitatively by ELISA. The contents of HYP in pulmonary tissues were analyzed quantitatively by sample hydrolysis method.

RESULTS

On the 14th and 28th days after treatment, the scores of pulmonary alveolitis and early fibrosis in pcDNA3.1-HGF plus BM-MSCs group were 2.36 ± 0.17, 2.8 ± 0.14 and 0.1 ± 0.11, 1.16 ± 0.13, which were significantly lower than those (1.68 ± 0.17, 1.58 ± 0.31 and 0.54 ± 0.15, 1.36 ± 0.13) in BM-MSCs group, also which were significantly lower those (2.36 ± 0.17, 2.80 ± 0.14 and 0.64 ± 0.09, 1.84 ± 0.17) in model group (P < 0.05); On the 14th and 28th days after treatment, the TNF-α contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 280.4 ± 23.11 and 249.78 ± 22.33 pg/mg, which were significantly lower than those (341.58 ± 35.34, 442.29 ± 36.76 pg/mg and 319.51 ± 17.84, 348.53 ± 33.95 pg/mg) in BM-MSCs and model groups (P < 0.05); On the 14th and 28th days after treatment, the HYP contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 0.46 ± 0.04 and 0.65 ± 0.05 µg/mg, which were significantly lower than those (0.63 ± 0.04, 1.04 ± 0.07 µg/mg and 0.72 ± 0.60, 1.39 ± 0.60 µg/mg) in BM-MSCs and model groups (P < 0.05).

CONCLUSION

The effects of BM-MSCs transfected by pcDNA3.1-HGF on suppressing pulmonary alveolitis and early fibrosis induced by SiO2 were better than those of BM-MSCs. The mechanism may be associated with the reduced pulmonary inflammation.

摘要

目的

比较骨髓间充质样干细胞(BM-MSCs)与经pcDNA3.1-HGF转染的BM-MSCs对二氧化硅诱导的肺泡炎及早期纤维化的影响差异,并探讨其作用机制。

方法

取雄性Wistar幼鼠的原代BM-MSCs进行培养,并用4,6-二脒基-2-苯基吲哚(DAPI)标记。50只Wistar大鼠随机分为3组:模型组(10只),气管内给予二氧化硅,次日经尾静脉注射PBS;BM-MSCs组(20只),气管内给予二氧化硅,次日经尾静脉注射1 ml BM-MSCs悬液;pcDNA3.1-HGF+BM-MSCs组(20只),气管内给予二氧化硅,次日经尾静脉注射1 ml经pcDNA3.1-HGF转染的BM-MSCs悬液。治疗后第14天和第28天,分别处死半数动物,取肺组织进行冰冻切片,在荧光显微镜下观察DAPI标记的细胞,在光学显微镜下进行HE染色,并用HE和Masson染色观察肺泡炎和纤维化情况。采用蛋白质印迹法检测大鼠肺组织中HGF的表达。用酶联免疫吸附测定法(ELISA)定量分析肺组织中肿瘤坏死因子-α(TNF-α)的表达水平。采用样品水解法定量分析肺组织中羟脯氨酸(HYP)的含量。

结果

治疗后第14天和第28天,pcDNA3.1-HGF+BM-MSCs组的肺泡炎和早期纤维化评分分别为2.36±0.17、2.8±0.14和0.1±0.11、1.16±0.13,显著低于BM-MSCs组(1.68±0.17、1.58±0.31和0.54±0.15、1.36±0.13),也显著低于模型组(2.36±0.17、2.80±0.14和0.64±0.09、1.84±0.17)(P<0.05);治疗后第14天和第28天,pcDNA3.1-HGF+BM-MSCs组肺组织中TNF-α含量分别为280.4±23.11和249.78±22.33 pg/mg,显著低于BM-MSCs组和模型组(341.58±35.34、442.29±36.76 pg/mg和319.51±17.84、348.53±33.95 pg/mg)(P<0.05);治疗后第14天和第28天,pcDNA3.1-HGF+BM-MSCs组肺组织中HYP含量分别为0.46±0.04和0.65±0.05 μg/mg,显著低于BM-MSCs组和模型组(0.63±0.04、1.04±0.07 μg/mg和0.72±0.6o、1.39±0.60 μg/mg)(P<0.05)。

结论

经pcDNA3.1-HGF转染的BM-MSCs对二氧化硅诱导的肺泡炎和早期纤维化的抑制作用优于BM-MSCs。其机制可能与减轻肺部炎症有关。

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