中草药

亚砷酸钠协同N-乙酰半胱氨酸对特发性肺纤维化的治疗作用

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袁一超, 李策, 陈楠, 熊域, 张纪平, 郑杭生, 朱志红

中草药 | 中药防治器官纤维化研究 2026,57(18): 7150-7161

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中草药 |中药防治器官纤维化研究 2026 , 57 (18) : 7150 -7161

亚砷酸钠协同N-乙酰半胱氨酸对特发性肺纤维化的治疗作用

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袁一超, 李策, 陈楠, 熊域, 张纪平, 郑杭生, 朱志红

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通讯作者:

郑杭生

作者简介:

袁一超: 袁一超（2002—），男，硕士研究生，研究方向为中药药剂学。E-mail:1944557087@qq.com#共同第一作者：李策，男，硕士研究生，研究方向为中药药剂学。E-mail:17367219539@163.com

Synergistic therapeutic efficacy of sodium arsenite and N-acetylcysteine against idiopathic pulmonary fibrosis

YUAN Yichao, LI Ce, CHEN Nan, XIONG Yu, ZHANG Jiping, ZHENG Hangsheng, ZHU Zhihong

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doi: 10.7501/j.issn.0253-2670.2026.18.007

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目的探究砒霜（三氧化二砷）的生物转化产物亚砷酸钠（sodium arsenite，SA）与N-乙酰半胱氨酸（N-acetylcysteine，NAC）联合用药的抗肺纤维化作用。方法小鼠L929成纤维细胞给予SA或NAC干预24 h后，测定细胞活力；通过考察不同比例的SA与NAC对L929细胞活力的影响，确定联合用药比例，采用划痕实验和Transwell实验检测细胞迁移能力；采用免疫荧光法检测SA与NAC联合给药对转化生长因子-β1（transforming growth factor-β1，TGF-β1）诱导的L929细胞α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、I型胶原蛋白α1（collagen type I α1，COL1A1）表达和活性氧（reactive oxygen species，ROS）水平的影响。建立博来霉素诱导的小鼠特发性肺纤维化模型，给予SA或NAC干预28 d，观察小鼠生存情况和体质量变化；采用苏木素-伊红（hematoxylin-eosin，HE）和Masson染色观察肺组织病理变化；检测支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中总蛋白含量、TGF-β1和肺组织羟脯氨酸（hydroxyproline，Hyp）水平；采用免疫荧光法检测肺组织α-SMA、COL1A1、CD86和CD206表达。结果 SA与NAC联合用药的协同效应与二者比例有关，当SA∶NAC＝2∶1时，可显著抑制L929细胞活力与迁移（P＜0.001），且优于单药组（P＜0.01、0.001）；SA与NAC（2∶1）联合给药显著下调TGF-β1诱导的L929细胞中α-SMA、COL1A1表达（P＜0.001），并降低ROS水平（P＜0.01）。体内实验结果显示，与模型组比较，SA与NAC（2∶1）联合给药可以延长小鼠的生存周期，增加体质量，减轻肺组织损伤和胶原沉积，降低BALF中总蛋白含量、TGF-β1及肺组织Hyp水平（P＜0.01、0.001），降低肺组织α-SMA、COL1A1表达（P＜0.001），抑制肺组织巨噬细胞M2型极化（P＜0.001），且对主要脏器无明显毒性作用。结论亚砷酸钠与NAC联合用药具有协同抗肺纤维化效果，为砷剂联合巯基供体治疗特发性肺纤维化提供了实验依据。

关键词

特发性肺纤维化 / 砒霜 / 亚砷酸钠 / N-乙酰半胱氨酸 / TGF-β1 / 巨噬细胞M2型极化 / 氧化应激

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Objective To investigate the antifibrotic effect of combined treatment with sodium arsenite (SA, biotransformation product of arsenic trioxide) and N-acetylcysteine (NAC) on pulmonary fibrosis. Methods L929 fibroblasts were treated with SA or NAC for 24 h before measuring cell viability. The combination drug ratio was determined by examining the effects of different ratios of SA and NAC on viability of L929 cells, and the cell migration ability was detected using scratch assay and Transwell assay. Immunofluorescence assay was used to detect the effects of combined administration of SA and NAC on the expressions of α-smooth muscle actin (α-SMA), collagen type I α1 (COL1A1), and reactive oxygen species (ROS) level in transforming growth factor-β1 (TGF-β1)-induced L929 cells. A mouse model of idiopathic pulmonary fibrosis was established induced by bleomycin, after intervention with SA or NAC for 28 d, the survival and body weight changes of mice were observed. Hematoxylin-eosin (HE) and Masson staining were used to observe pathological changes in lung tissue. The total protein content, TGF-β1 level in bronchoalveolar lavage fluid (BALF), and hydroxyproline (Hyp) level in lung tissue were detected. Immunofluorescence assay was used to detect the expressions of α-SMA, COL1A1, CD86 and CD206 in lung tissue. Results The synergistic effect of SA and NAC combination therapy was related to their ratio. When SA∶NAC = 2∶1, the combination could significantly inhibit viability and migration of L929 cells (P < 0.001), and was superior to the monotherapy group (P < 0.01, 0.001). The combined administration of SA and NAC (2∶1) significantly down-regulated the expressions of α-SMA and COL1A1 in L929 cells induced by TGF-β1 (P < 0.001), and reduced ROS level (P < 0.01). The in vivo experimental results showed that compared with model group, the combined administration of SA and NAC (2∶1) could prolong the survival cycle of mice, increase body weight, reduce lung tissue damage and collagen deposition, decrease the total protein content, TGF-β1 level in BALF and Hyp level in liver tissue (P < 0.01, 0.001), reduce the expression of α-SMA and COL1A1 in lung tissue (P < 0.001), inhibit M2 polarization of lung tissue macrophages (P < 0.001), and had no significant toxic effects on major organs. Conclusion The combination of SA and NAC has a synergistic anti-pulmonary fibrosis effect, providing experimental evidence for the treatment of idiopathic pulmonary fibrosis with arsenic agents combined with thiol donors.

Key words

idiopathic pulmonary fibrosis / arsenic trioxide / sodium arsenite / N-acetylcysteine / TGF-β1 / M2 macrophage polarization / oxidative stress

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袁一超, 李策, 陈楠, 熊域, 张纪平, 郑杭生, 朱志红. 亚砷酸钠协同N-乙酰半胱氨酸对特发性肺纤维化的治疗作用. 中草药, 2026 , 57 (18) : 7150 -7161 . DOI: 10.7501/j.issn.0253-2670.2026.18.007

YUAN Yichao, LI Ce, CHEN Nan, XIONG Yu, ZHANG Jiping, ZHENG Hangsheng, ZHU Zhihong. Synergistic therapeutic efficacy of sodium arsenite and N-acetylcysteine against idiopathic pulmonary fibrosis[J]. Chinese Traditional and Herbal Drugs, 2026 , 57 (18) : 7150 -7161 . DOI: 10.7501/j.issn.0253-2670.2026.18.007

基金

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浙江省自然科学基金资助项目 (LY23H280010); 浙江中医药大学校级科研基金资助项目 (2023JKZKTS23)

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Zheng D B, Guo J S, Liang Z Y, et al. Supramolecular nanofibers ameliorate bleomycin-induced pulmonary fibrosis by restoring autophagy[J]. Adv Sci, 2024, 11(28):e2401327.
Zhang G R, Luo L, Zhang L M, et al. Research progress of respiratory disease and idiopathic pulmonary fibrosis based on artificial intelligence[J]. Diagnostics, 2023,13(3):357.
马鑫来,刘学,张伟.“肺虚络瘀,肺络癥瘕”病机观与系统性硬化症合并肺间质纤维化的相关性[J]. 安徽中医药大学学报, 2023, 42(4):8-11.
庄宛滢,晏军,李艳华,等.从“肺络微型癥瘕”理论探讨特发性肺纤维化治疗的“三期分治”原则[J]. 中华中医药杂志, 2023, 38(12):6100-6102.
王克俭.三氧化二砷治疗动物肺纤维化的深入研究[D]. 苏州:苏州大学, 2009.
高淑艳. As2O3上调miR-98表达缓解博莱霉素诱导的大鼠肺纤维化[D]. 滨州:滨州医学院, 2014.
Joannes A, Morzadec C, Duclos M, et al. Arsenic trioxide inhibits the functions of lung fibroblasts derived from patients with idiopathic pulmonary fibrosis[J]. Toxicol Appl Pharmacol, 2022, 441:115972.
Luo F Y, Zhuang Y, Sides M D, et al. Arsenic trioxide inhibits transforming growth factor-β1-induced fibroblast to myofibroblast differentiation in vitro and bleomycin induced lung fibrosis in vivo[J]. Respir Res, 2014, 15(1):51.
Zhong L Z, Hao H J, Chen D Y, et al. Arsenic trioxide inhibits the differentiation of fibroblasts to myofibroblasts through nuclear factor erythroid 2-like 2(NFE2L2)protein and the Smad2/3 pathway[J]. J Cell Physiol, 2019, 234(3):2606-2617.
Gao S Y, Zhou X, Li Y J, et al. Arsenic trioxide prevents rat pulmonary fibrosis via miR-98 overexpression[J]. Life Sci, 2014, 114(1):20-28.
Hu Y X, Li J, Lou B, et al. The role of reactive oxygen species in arsenic toxicity[J]. Biomolecules, 2020, 10(2):240.
Samarakoon R, Overstreet J M, Higgins P J. TGF-βsignaling in tissue fibrosis:Redox controls, target genes and therapeutic opportunities[J]. Cell Signal, 2013, 25(1):264-268.
Liu R M, Desai L P. Reciprocal regulation of TGF-β and reactive oxygen species:A perverse cycle for fibrosis[J]. Redox Biol, 2015, 6:565-577.
Rogliani P, Manzetti G M, Gholamalishahi S, et al. Impact of N-acetylcysteine on mucus hypersecretion in the airways:A systematic review[J]. Int J Chron Obstruct Pulmon Dis, 2024, 19:2347-2360.
Santus P, Signorello J C, Danzo F, et al. Anti-inflammatory and anti-oxidant properties of N-acetylcysteine:A fresh perspective[J]. J Clin Med, 2024, 13(14):4127.
Rhee C K, Lim S Y, Lee W Y, et al. The effect of nebulized N-acetylcysteine on the phlegm of chronic obstructive pulmonary disease:The NEWEST study[J]. BMC Pulm Med, 2024, 24(1):434.
Mokra D, Mokry J, Barosova R, et al. Advances in the use of N-acetylcysteine in chronic respiratory diseases[J]. Antioxidants, 2023, 12(9):1713.
Ralph S J. Arsenic-based antineoplastic drugs and their mechanisms of action[J]. Met Based Drugs, 2008, 2008:260146.
Jin Z, Yi X, Yang J, et al. Liposome-coated arsenicmanganese complex for magnetic resonance imagingguided synergistic therapy against carcinoma[J]. Int J Nanomed, 2021, 16:3775-3788.
Ettlinger R, Moreno N, Volkmer D, et al. Zeolitic imidazolate framework-8 as pH-sensitive nanocarrier for“arsenic trioxide” drug delivery[J]. Chem A Eur J, 2019,25(57):13189-13196.
Fu X, Liang Q R, Luo R G, et al. An arsenic trioxide nanoparticle prodrug(ATONP)potentiates a therapeutic effect on an aggressive hepatocellular carcinoma model via enhancement of intratumoral arsenic accumulation and disturbance of the tumor microenvironment[J]. J Mater Chem B, 2019, 7:3088-3099.
Jiao H, Song J, Sun X, et al. Sodium arsenite inhibits lung fibroblast differentiation and pulmonary fibrosis[J]. Pharmacol, 2019, 104(5/6):368-376.
Ganapathy S, Xiao S, Yang M, et al. A low-dose arsenicinduced p53 protein-mediated metabolic mechanism of radiotherapy protection[J]. J Biol Chem, 2014, 289(8):5340-5347.
Ganapathy S, Xiao S, Seo S J, et al. Low-dose arsenic induces chemotherapy protection via p53/NF-κBmediated metabolic regulation[J]. Oncogene, 2014,33(11):1359-1366.
Chung J, Huda M N, Shin Y, et al. Correlation between oxidative stress and transforming growth factor-beta in cancers[J]. Int J Mol Sci, 2021, 22(24):13181.
Hecker L, Vittal R, Jones T, et al. NADPH oxidase-4mediates myofibroblast activation and fibrogenic responses to lung injury[J]. Nat Med, 2009, 15(9):1077-1081.
Xu S W, Thompson K, Khan K, et al. Focal adhesion kinase and reactive oxygen species contribute to the persistent fibrotic phenotype of lesional scleroderma fibroblasts[J]. Rheumatology, 2012, 51(12):2146-2154.
Cantin A M, Hubbard R C, Crystal R G. Glutathione deficiency in the epithelial lining fluid of the lower respiratory tract in idiopathic pulmonary fibrosis[J]. Am Rev Respir Dis, 1989, 139(2):370-372.
Pommerolle L, Beltramo G, Biziorek L, et al. CD206+macrophages are relevant non-invasive imaging biomarkers and therapeutic targets in experimental lung fibrosis[J]. Thorax, 2024, 79(12):1124-1135.
Ghebremedhin A, Salam A B, Adu-Addai B, et al. A novel CD206 targeting peptide inhibits bleomycin-induced pulmonary fibrosis in mice[J]. Cells, 2023, 12(9):1254.
Zhang Y, Choksi S, Chen K, et al. ROS play a critical role in the differentiation of alternatively activated macrophages and the occurrence of tumor-associated macrophages[J]. Cell Res, 2013, 23(7):898-914.
Zhu Q Y, Tai S, Tang L, et al. N-acetyl cysteine ameliorates aortic fibrosis by promoting M2 macrophage polarization in aging mice[J]. Redox Rep, 2021, 26(1):170-175.
Yin J Z, Shi X Q, Wang M D, et al. Arsenic trioxide elicits anti-tumor activity by inhibiting polarization of M2-like tumor-associated macrophages via Notch signaling pathway in lung adenocarcinoma[J]. Int Immunopharmacol, 2023, 117:109899.
Hung C H, Hsu H Y, Chiou H C, et al. Arsenic induces M2macrophage polarization and shifts M1/M2 cytokine production via mitophagy[J]. Int J Mol Sci, 2022, 23(22):13879.

2026年第57卷第18期

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doi: 10.7501/j.issn.0253-2670.2026.18.007

接收时间：2026-01-22
首发时间：2026-10-29

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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