金水缓纤方通过p53-KRT8信号通路减轻HPAEPICS细胞衰老和分化障碍改善肺纤维化的作用机制
Mechanism of Jinshui Huanxian Formula on improving pulmonary fibrosis by alleviating senescence and dysdifferentiation of HPAEPIC cells via p53-KRT8 signaling pathway
摘要
Abstract
Objective:To explore the effects and mechanisms of Jinshui Huanxian Formula on improving bleomycin(BLM)-induced pulmonary fibrosis by alleviating type Ⅱ alveolar epithelial cells HPAEPIC senescence and dysdifferentiation via regulating the p53-KRT8 signaling pathway.Methods:HPAEPIC cells were induced by 0.63 µg/mL BLM for 48 h to establish a cellular pulmonary fibrosis model.The cells were divided into the blank group(CON group),the model group(MOD group),the Jinshui Huxian Formula group(JHF group,10%),the pirfenidone group(PFD group,20 µg/mL)and the p53 inhibitor group(pifithrin-α HBr inhibitor,PFT-α group,10 µmol/L).Cell viability was detected by CCK-8 assay.The mRNA expression levels of IL-6,IL-8,p21,p53,SPC,KRT8,and IL-13 were detected by RT-PCR.The cell cycle was detected by flow cytome-try.The expression levels of p53,KRT8,CLDN4,N-CAD,and α-SMA were detected by Western blot.The expression levels of p53,p21,and KRT8 was detected by immunofluorescence.Results:Compared to the CON group,the level of G1 phase per-centage and G1/G2 ratio in the MOD group were significantly decreased(P<0.01),the level of G2 phase percentage was signifi-cantly increased(P<0.01).The mRNA expression levels of IL-6,IL-8,p21,p53,KRT8,and CLDN4 were significantly up-regulated(P<0.05).The protein expression levels of p53,KRT8,CLDN4,N-CAD,and α-SMA were significantly in-creased(P<0.05).The fluorescence expression levels of p53,p21,and KRT8 were significantly increased(P<0.01).Com-pared to the MOD group,the levels of G1 phase percentage in the JHF and PFT-α groups were significantly increased(P<0.05),and the levels of G2 phase percentage was significantly decreased(P<0.01).The mRNA expression levels of IL-6,IL-8,p21,p53,KRT8,and CLDN4 were significantly down-regulated(P<0.05).The protein expression levels of p53,KRT8,CLDN4,N-CAD,and α-SMA were significantly reduced(P<0.05).The fluorescence expression levels of p53,p21,and KRT8 were significantly decreased(P<0.01).The Pearson's coefficient between KRT8 fluorescence and p21 fluorescence in all groups was>0.5.Conclusion:Jinshui Huanxian Formula can improve BLM-induced HPAEPIC cell pulmonary fibrosis,and its mecha-nism may be related to the alleviation of HPAEPIC cell senescence and dysdifferentiation via inhibiting the p53-KRT8 signaling pathway.关键词
肺纤维化/衰老/金水缓纤方/KRT8/Ⅱ型肺泡上皮细胞Key words
Pulmonary fibrosis/Senescence/Jinshui Huanxian Formula/KRT8/Type Ⅱ alveolar epithelial cells分类
医药卫生引用本文复制引用
王宇,游梦月,余本嫚,陈媛媛,韩瑞婷,沈俊岭,余海滨..金水缓纤方通过p53-KRT8信号通路减轻HPAEPICS细胞衰老和分化障碍改善肺纤维化的作用机制[J].海南医科大学学报,2026,32(4):258-268,11.基金项目
This study was supported by the National Key Research Project for the Modernization of Traditional Chinese Medicine(2023YFC3502604) （2023YFC3502604）
Natural Science Foundation Project of Henan Province(222300420221) （222300420221）
Major Special Project of Traditional Chinese Medicine Research in Henan Province(2022ZYZD04) （2022ZYZD04）
Scientific Research Program of Traditional Chinese Medicine in Henan Province(2021JDZY102,2023ZXZX1033) （2021JDZY102,2023ZXZX1033）
Special Project of Collaborative Innovation Center of Zhengzhou(2023XTCX043) 国家重点研究计划中医药现代化重点专项(2023YFC3502604) （2023XTCX043）
河南省自然科学基金(222300420221) （222300420221）
河南省中医药研究重大专项(2022ZYZD04) （2022ZYZD04）
河南省中医药科学研究专项(2021JDZY102,2023ZXZX1033) （2021JDZY102,2023ZXZX1033）
郑州市协同创新中心专项(2023XTCX043) （2023XTCX043）
