摘要
目的探讨槲皮素对肺纤维化大鼠肺组织细胞间质转化、炎症及对NOX4-P62信号途径的影响,为进一步阐明槲皮素对肺纤维化的作用机制提供实验依据。方法将实验大鼠分为对照组、博来霉素组(BLM组)和博来霉素+槲皮素组(BLM+QUE组),分别于造模后第7、14、28天观察各组肺组织外观,利用苏木素-伊红染色法(HE染色)评价肺纤维化情况,蛋白质免疫印迹法(Western blot)检测各组肺组织还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)、自噬蛋白P62、核因子κB(NF-κB)的蛋白表达情况,反转录-聚合酶链反应(qRT-PCR)检测人肺腺癌细胞株A549细胞血管平滑肌肌动蛋白(α-SMA)和钙粘附蛋白E(E-cadherin)的mRNA表达水平,酶联免疫吸附法(ELISA法)检测转化生长因子-β1(TGF-β1)、白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的水平。结果BLM组随时间增加大鼠肺纤维化呈进行性加重,BLM+QUE组在第7天时症状较BLM组轻,在第28天时症状明显减轻。BLM组的TGF-β1、IL-1β、TNF-α、NOX4、P62、NF-κB蛋白表达,较对照组明显上升,差异具有统计学意义(P<0.05),BLM+QUE组在第7天时较BLM组低,但无显著差异(P>0.05),但在第14、28天时较BLM组明显下降,差异显著(P<0.05)。与对照组比较,加入TGF-β1的A549细胞E-cadherin、α-SMA mRNA的表达分别明显降低和升高(P<0.05);给予槲皮素后,E-cadherin、α-SMA mRNA的表达较TGF-β1组分别明显升高和降低(P<0.05)。结论槲皮素能够抑制TGF-β1对人肺腺癌细胞株A549细胞诱导的间质转化的转变,减轻肺纤维化大鼠的炎症反应,缓解博来霉素所致的大鼠肺纤维化,自噬活性在博来霉素诱导大鼠发生肺纤维化的过程中受到抑制;槲皮素可能通过抑制NOX4-P62信号途径,激活自噬,减轻大鼠肺纤维化。
Objective To investigate the effects of quercetin on interstitial transformation, inflammation, and the NOX4-P62 signaling pathway in lung tissue of rats with pulmonary fibrosis to provide an experimental basis for further elucidating the mechanism of quercetin in pulmonary fibrosis. Methods Rats were divided into control, BLM and BLM+QUE groups. The appearance of lung tissue in each group was observed on days 10, 20 and 30 after modeling. Pulmonary fibrosis was evaluated by HE staining. NOX4 and P62 in lung tissue, and α-SMA and E-cadherin in A549 cells were detected by western blotting. TGF-β1, IL-1β and TNF-α were measured by ELISA. Results Symptoms in the BLM+QUE group were milder than those in the BLM group on day 7, and significantly relieved on day 28. TGF-β1, IL-1β, TNF-α, NOX4 and P62 expression in the BLM+QUE group was lower than that in the BLM group on day 10(P>0.05), but it was much expression lower than that in the BLM group on days 14 and 28(P<0.05). Compared with the control group, E-cadherin and α-SMA expression in the TGF-β1 group was significantly decreased and increased, respectively(P<0.05). Compared with the TGF-β1 group, E-cadherin and α-SMA expression in the TGF-β1+QUE group were significantly increased and decreased, respectively(P<0.05). Conclusions Quercetin inhibits TGF-β1. EMT of A549 cells alleviates the inflammatory reaction and pulmonary fibrosis induced by bleomycin in rats. Autophagy is inhibited by bleomycin-induced pulmonary fibrosis in rats. Quercetin inhibits the NOX4-P62 signaling pathway, activates autophagy, and reduces pulmonary fibrosis in rats.
作者
薛兰
毛春迎
王慧
王守福
张争辉
郭新庆
XUE Lan;MAO Chunying;WANG Hui;WANG Shoufu;ZHANG Zhenghui;GUO Xinqing(Department of basic Medicine,Heze Medical College,Heze 274000,China;Department of Medical Technology,Heze Medical College,Heze 274000)
出处
《中国比较医学杂志》
CAS
北大核心
2022年第7期94-100,117,共8页
Chinese Journal of Comparative Medicine
