Inhibition of miR21-mediated Ang (1-7) on activation of AngⅡ-induced inflammatory body in pulmonary fibroblasts

doi:10.3877/cma.j.issn.1674-6902.2019.05.012

Abstract

Abstract:

Objective

To investigate the effects of miR21 on the proliferation and apoptosis of pulmonary fibroblasts and the possible mechanisms.

Methods

MiR21 NC (miR21 NC group), the inhibitor (inhibitor group), the mimics (mimics group), and nothing (blank control group) were transfected into the primary lung fibroblasts, respectively. The effects of miR21 on the proliferation of lung fibroblasts were detected by CCK-8 cell proliferation. The effects of miR21 on the apoptosis of pulmonary fibroblasts were detected by flow cytometry. The expression levels of miR21 and Ang (1-7) in the pulmonary fibroblasts were detected by reverse transcription polymerase chain reaction (RT-PCR) before and after transfection. The expression levels of miR21 and NLRP3 were detected before and after transfection by Western blotting method.

Results

According to the results of CCK8 and flow cytometry detection, miR21 can significantly promote the proliferation of pulmonary fibroblasts and inhibit the apoptosis of pulmonary fibroblasts. RT-PCR analysis showed that after the success of cell transfection, the miR21 mimics group and the inhibitor group respectively inhibited/promoted Ang (1-7) expression and promoted/inhibited Ang Ⅱ expression (P<0.05). That is to say, miR21 could inhibit miR21 Ang (1-7) and promote Ang Ⅱ expression. Western blot analysis showed that the miR21 mimic/inhibitor could raise/lower the expression levels of Ang Ⅱ and the expression levels of NLRP3 protein, which had statistical significant difference compared with the NC control groups (P<0.05).

Conclusion

MiR21 can promote the proliferation and inhibit the apoptosis of pulmonary fibroblasts, and its mechanism may be related to the effect of miR21-mediated Ang (1-7) on the activation of AngⅡ-induced inflammatory body in pulmonary fibroblasts.

Key words: MiR21, Pulmonary fibrosis, Proliferation, Apoptosis, Mechanism of action

Guangfu Shen, Pei Du, Rui Yu, Zhaofeng Xie, Changqin Luo. Inhibition of miR21-mediated Ang (1-7) on activation of AngⅡ-induced inflammatory body in pulmonary fibroblasts[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2019, 12(05): 596-600.

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https://zhfbjbzz.cma-cmc.com.cn/EN/Y2019/V12/I05/596

Figures/Tables 8

References 20

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指　标	Vimentin(灰度值)
正常肺泡细胞	0.74±0.06
肺成纤维细胞	3.31±0.72
F值	4.06
P值	<0.05

指　标	Vimentin(灰度值)
正常肺泡细胞	0.74±0.06
肺成纤维细胞	3.31±0.72
F值	4.06
P值	<0.05

组　别	肺成纤维细胞OD值
组　别	24 h	48 h	72 h
空白对照组	0.82±0.05	0.86±0.06	0.91±0.01
miR21 NC组	0.82±0.03^a	0.85±0.01^a	0.91±0.03^a
inhibitor组	0.81±0.04^a	0.83±0.06^a	0.86±0.07^a
mimic组	0.82±0.03^a	0.91±0.02^a	0.96±0.02^a
F值	8.252	13.706	11.485
P值	>0.05	<0.05	<0.05

组　别	肺成纤维细胞OD值
组　别	24 h	48 h	72 h
空白对照组	0.82±0.05	0.86±0.06	0.91±0.01
miR21 NC组	0.82±0.03^a	0.85±0.01^a	0.91±0.03^a
inhibitor组	0.81±0.04^a	0.83±0.06^a	0.86±0.07^a
mimic组	0.82±0.03^a	0.91±0.02^a	0.96±0.02^a
F值	8.252	13.706	11.485
P值	>0.05	<0.05	<0.05

组　别	肺成纤维细胞凋亡率(%)
组　别	24 h	48 h	72 h
空白对照组	26.24±4.20	35.27±4.37	39. 25±4.61
miR21 NC组	28.20±5.27^a	35.23±4.32^a	38.34±5.57^a
inhibitor组	42.91±3.76^a	51.27±4.21^a	68.94±4.21^a
mimic组	15.23±3.06^a	18.72±4.24^a	23.63±4.04^a
F值	10.814	12.922	10.723
P值	<0.05	<0.05	<0.05

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