海南医科大学学报2025，Vol.31Issue(15)：1153-1162,10.DOI:10.13210/j.cnki.jhmu.20240911.002
金水缓纤方调控CD38/NAD+-SIRT1轴延缓AEC2s细胞衰老抗肺纤维化的机制研究
Mechanism of Jinshui Huanxian Formula in regulating CD38/NAD+-SIRT1 axis to delay AEC2s cell senescence and alleviate-pulmonary fibrosis
摘要
Abstract
Objective:To investigate the mechanism of Jinshui Huanxian Formula(JHF)regulating AEC2s cell senescence and alleviating-pulmonary fibrosis through CD38/NAD+-SIRT1 axis.Methods:SD rats were randomly divided into a blank con-trol group,a model group,a JHF group,a positive control group,and a CD38 inhibitor group,with 10 rats in each group.A rat model of pulmonary fibrosis was induced by intratracheal instillation of bleomycin.On the 28th day after modeling,the rats were treated with intragastric administration of drugs,once a day for 14 consecutive days.H&E and Masson staining were used to ob-serve and evaluate the degree of inflammation and fibrosis in lung tissue.The protein expression of αSMA and COLⅢ in lung tis-sue was evaluated by immunohistochemistry.Immunofluorescence was used to detect the expression of p21 and SPC in lung tis-sue.Cell senescence in lung tissue was determined by β-galactosidase(SA-β-gal)staining.The mRNA levels of IL-1α,IL-8,P53,P21,CD38,and SIRT1 were detected by QPCR.Western Blot was used to measure the protein expression of E-cad,N-cad,p21,p53,CD38,and SIRT1 in lung tissue.The expression of NAD+in lung tissue was detected by WST-8 method.Re-sults:Compared to the blank group,the inflammation and fibrosis scores in the lung tissue of the model group were significantly in-creased.The protein expressions of αSMA and COLⅢ were significantly increased,suggesting the occurrence of lung fibrosis in rats.The expression of N-cad protein in lung tissue was significantly up-regulated,and the expression of E-cad protein was de-creased,suggesting that epithelial-mesenchymal transition occurred in lung tissue.SA-β-gal blue staining,p53 and p21 mRNA and protein expression,and IL-1α and IL-8 mRNA expression were significantly increased in lung tissue.Senescent cells accumu-lated and highly expressed senescence-associated secretory phenotype in lung tissue.Immunofluorescence results showed that p21 was co-expressed with SPC,and p21 was highly expressed in AEC2s.The expression of CD38 mRNA and protein was signifi-cantly increased,the expression of SIRT1 mRNA and protein was significantly decreased,and the expression of NAD+was signif-icantly decreased,suggesting that the consumption of NAD+may be accompanied by pulmonary fibrosis.Compared to the model group,the inflammation and fibrosis scores in the lung tissue of the treatment groups were significantly decreased,and the expres-sion of αSMA and COLⅢ protein was significantly decreased.The expression of N-cad protein was significantly down-regulated and E-cad protein was increased.SA-β-gal blue staining in lung tissue was significantly reduced,the degree of cell senescence was significantly improved,and the expression levels of senescence-related genes and proteins of p53,p21,IL-1α,and IL-8 were sig-nificantly down-regulated.The expression of CD38 mRNA and protein decreased,while the expression of SIRT1 and tissue NAD+increased.Conclusion:Jinshui Huanxian Formula can reduce bleomycin-induced pulmonary fibrosis and inflammatory re-sponse in lung tissue of rats,improve collagen deposition,which may be related to regulating CD38 expression,increasing NAD+level,activating SIRT1 protein,and delaying AEC2s cell senescence.关键词
金水缓纤方/肺纤维化/细胞衰老/CD38/NAD+-SIRT1/Ⅱ型肺泡上皮细胞Key words
Jinshui Huanxian Formula/Pulmonary fibrosis/Cellular senescence/CD38/NAD+-SIRT1/Type Ⅱ alveolar epithelial cells分类
医药卫生引用本文复制引用
蔡宇,王宇,沈俊岭,游梦月,韩瑞婷,余海滨..金水缓纤方调控CD38/NAD+-SIRT1轴延缓AEC2s细胞衰老抗肺纤维化的机制研究[J].海南医科大学学报,2025,31(15):1153-1162,10.基金项目
This study was supported by the Major Special Project of Traditional Chinese Medicine Research in Henan Province(2022ZYZD04),Special Project of Collaborative Innovation Center of Zhengzhou(2023XTCX043) （2022ZYZD04）
Natural Science Foundation of Henan Province(222300420221)and Scientific Research Program of Traditional Chinese Medicine in Henan Province(2021JDZY102) 河南省中医药研究重大专项资助项目(2022ZYZD04) （222300420221）
郑州市协同创新中心专项资助项目(2023XTCX043) （2023XTCX043）
河南省自然科学基金资助项目(222300420221) （222300420221）
河南省中医药科学研究专项资助项目(2021JDZY102) （2021JDZY102）
