Extract of Qixian Decoction inhibits airway epithelial-mesenchymal transition in asthma

TANG Lingling; NI Zhenhua; CHEN Qingge; MENG Ziyu; WANG Xiongbiao

doi:10.16305/j.1007-1334.2018.12.021

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Extract of Qixian Decoction inhibits airway epithelial-mesenchymal transition in asthma

更新时间：2022-08-31

- Extract of Qixian Decoction inhibits airway epithelial-mesenchymal transition in asthma
- Shanghai Journal of Traditional Chinese Medicine Vol. 52, Issue 12, Pages: 78-83(2018)
- 作者机构：
  
  1. 上海中医药大学附属普陀医院呼吸内科,上海,200062
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2018.12.021
  CLC：
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TANG Lingling, NI Zhenhua, CHEN Qingge, et al. Extract of Qixian Decoction inhibits airway epithelial-mesenchymal transition in asthma. [J]. Shanghai Journal of Traditional Chinese Medicine 52(12):78-83(2018)
DOI：

TANG Lingling, NI Zhenhua, CHEN Qingge, et al. Extract of Qixian Decoction inhibits airway epithelial-mesenchymal transition in asthma. [J]. Shanghai Journal of Traditional Chinese Medicine 52(12):78-83(2018) DOI： 10.16305/j.1007-1334.2018.12.021.

摘要

目的：以16 HBE人支气管上皮细胞株为研究对象,通过体外研究探讨芪仙汤提取物对哮喘气道上皮间质转化的影响。方法：将16HBE细胞加入不同浓度的H,2,O,2,及芪仙汤提取物干预不同时间。观察细胞形态改变；CCK-8检测H,2,O,2,对16HBE细胞活力的影响；RT-PCR检测E-cadherin、α-SMA基因表达水平；Western blot检测E-cadherin、α-SMA蛋白表达水平；免疫荧光检测E-cadherin,α-SMA蛋白表达水平及蛋白定位。结果：H,2,O,2,作用后细胞形态呈EMT改变,而芪仙汤提取物可逆转细胞EMT改变；CCK-8结果显示,H,2,O,2,作用后气道上皮细胞存活率随浓度增加而降低(P,<,0.01),芪仙汤提取物可以减少H,2,O,2,诱导的气道上皮细胞损伤(P,<,0.01)。RT-PCR检测结果示,H,2,O,2,刺激后气道上皮细胞E-cadherin mRNA水平显著降低(P,<,0.01),α-SMA mRNA水平增高(P,<,0.05),芪仙汤提取物作用后可以逆转H,2,O,2,诱导的E-cadherin转录水平降低(P,<,0.05),抑制H,2,O,2,诱导的α-SMA mRNA表达水平增加(P,<,0.05)。Western blot及免疫荧光检测结果示E-cadherin、α-SMA蛋白表达水平与基因表达水平相一致(P,<,0.05)。结论：芪仙汤提取物可以减少氧化应激诱导的气道上皮细胞损伤及气道上皮间质转化。

Abstract

Objective:Taken 16HBE human bronchial epithelial cell line as the research object, the effects of Qixian Decoction extract on inhibiting airway epithelial-mesenchymal transition(EMT)in asthma are investigated in vitro. MethodsThe 16HBE cells were intervened by H,2,O,2, and Qixian Decoction extract at different concentrations for different time to observe the cell morphology changes. CCK-8 assay was used to detect the effect of H,2,O,2, on the viability of 16HBE cells. RT-PCR was used to detect the gene expression levels of E-cadherin and α-SMA. The protein expression levels of E-cadherin and α-SMA were detected by Western blot. The protein expression levels and protein localization of E-cadherin and α-SMA were detected by immunofluorescence. Results:The morphology of cells showed the changes of EMT after H,2,O,2, treatment, and the extract of Qixian Decoction reversed the EMT changes. The results of CCK-8 assay showed that the survival rate of airway epithelial cells was decreased with the increasing concentration of H,2,O,2, (P,<,0.01). The extract of Qixian Decoction reduced the damage of airway epithelial cells induced by H,2,O,2, (P,<,0.01). The results of RT-PCR showed that the level of E-cadherin mRNA in airway epithelial cells was significantly decreased after H,2,O,2, stimulation (P,<,0.01), and the level of α-SMA mRNA was increased (P,<,0.05). The extract of Qixian Decoction reversed the decreased level of E-cadherin transcription induced by H,2,O,2, (P,<,0.05), as well as the increased expression of α-SMA mRNA induced by H,2,O,2, (P,<,0.05). The results of Western blot and immunofluorescence showed that the protein expression levels of E-cadherin and α-SMA were consistent with the gene expression levels (P,<,0.05). Conclusion:Qixian Decoction can reduce the airway epithelial cell damage and airway epithelial-mesenchymal transition induced by oxidative stress.

关键词

芪仙汤气道上皮间质转化氧化应激

Keywords

Qixian Decoctionairway epithelial-mesenchymal transitionoxidative stress

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