延龄草皂苷通过抑制TGF-β/Smad3与Wnt/β-catenin信号通路改善大鼠肺纤维化

doi:10.6040/j.issn.1671-7554.0.2022.0011

摘要/Abstract

摘要： 目的探讨延龄草皂苷对肺间质纤维化大鼠的治疗作用及其可能机制。方法雄性SD大鼠50只,随机分为对照组,模型组,延龄草皂苷低、中、高剂量组［25 mg/(kg·d)、50 mg/(kg·d)和100 mg/(kg·d)］,每组10只。除对照组外,其余各组采用气管插管注入伯莱霉素法(5 mg/kg)建立大鼠肺纤维化模型;次日开始连续给药28 d。处死大鼠后,取肺组织称重并计算肺系数;进行HE和Masson染色观察小鼠肺组织形态病理学变化;ELISA法检测大鼠白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)、丙二醛(MDA)和活性氧(ROS)的表达以及超氧化物歧化酶(SOD)的活性;Western blotting法检测大鼠肺组织中纤连蛋白(fibronectin)、胶原蛋白Ⅰ型(Collagen Ⅰ)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β(TGF-β)、SMAD同源物3(Smad3)、Wnt3a、β-连环蛋白(β-catenin)和糖原合成酶激酶-3β(GSK-3β)的蛋白表达水平。结果与模型组比较,延龄草皂苷低、中、高剂量组的肺系数降低;大鼠肺组织炎性细胞浸润、胶原沉积和纤维化程度改善;大鼠IL-1β、TNF-α、IL-6、MDA和ROS的表达水平降低, SOD的活性升高,发挥抗炎、抗氧化作用;肺纤维化标志蛋白fibronectin,Collagen Ⅰ和α-SMA表达被抑制;大鼠肺组织TGF-β1、Smad3、Wnt3a、β-catenin和GSK-3β蛋白的表达水平降低,且治疗效果呈现剂量依赖性。结论延龄草皂苷可通过抗炎、抗氧化,抑制TGF-β/Smad3与Wnt/β-catenin信号通路激活,从而对肺纤维化产生保护作用。

关键词: 延龄草皂苷, 肺纤维化, 炎症, 转化生长因子-β/ SMAD同源物3信号通路, Wnt/β-catenin信号通路

Abstract: Objective To investigate the treatment and mechanism of Trillium Saponins in rats with pulmonary interstitial fibrosis. Methods Fifty male SD rats were divided into 5 groups, namely the control group, the model group, and the low, medium, and high Trillium Saponins treatment groups, with 10 in each group. Except the control group, the other groups were injected with bleomycin 5 mg/kg through tracheal intubation. A rat model of pulmonary fibrosis was established. The treatment group was given 25, 50 and 100 mg/(kg·d)Trillium Saponins by gavage the next day. After 28 days of treatment, the rats were sacrificed, and blood and lung tissues were taken. The lung tissue was weighed and the lung coefficient was calculated. HE staining and Masson staining were used to observe rat lung histopathology. The expression of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), malondialdehyde(MDA)and reactive oxygen species(ROS)and the activity of superoxide dismutase(SOD)were detected by ELISA kit. Western blotting experiment was employed to detect the protein expression levels of transforming growth factor-β(TGF-β)1, Smad3, Wnt3a, β-catenin, glycogen synthase kinase-3β(GSK-3β), fibronectin, Collagen Ⅰ and α-SMA in lung tissue. Results Compared with the model group, lung coefficient was decreased, inflammatory cell infiltration, collagen deposition and fibrocytosis was reduced, the expression of IL-1β, TNF-α, IL-6, MDA and ROS levels was decrease, the activity of SOD was increased, the expression of fibronectin, Collagen Ⅰ, α-SMA, TGFβ1, Smad3, Wnt3a, β-catenin and GSK-3β proteins was inhibited in lung tissue of low, medium, and high Trillium Saponins treatment groups. Conclusion Trillium has anti-inflammatory and anti-oxidant properties, and can inhibit the activation of TGF-β/Smad3 and Wnt/β-catenin signaling pathways to protect against pulmonary fibrosis.

Key words: Trillium saponin, Pulmonary interstitial fibrosis, Inflammation, Transforming growth factor-β/Smad3 signaling pathway, Wnt/β-catenin signaling pathway

中图分类号:

R285.5

张秉芬,周胜红,王哲. 延龄草皂苷通过抑制TGF-β/Smad3与Wnt/β-catenin信号通路改善大鼠肺纤维化[J]. 山东大学学报 (医学版), 2022, 60(8): 23-29.

ZHANG Bingfen, ZHOU Shenghong, WANG Zhe. Trillium Saponins ameliorates pulmonary fibrosis in rats by inhibiting TGF-β/Smad3 and Wnt/β-catenin signaling pathways[J]. Journal of Shandong University (Health Sciences), 2022, 60(8): 23-29.

参考文献

[1] Herrera J, Henke CA, Bitterman PB. Extracellular matrix as a driver of progressive fifibrosis[J]. J Clin Invest, 2018, 128(1): 45-53.
[2] 伍徐娴, 杨勤, 罗新华, 等. 芍化纤胶囊对肝细胞线粒体脂质过氧化的影响 [J].贵阳医学院学报, 2004, 29(5): 406-409. WU Xuxian, YANG Qin, LUO Xinhua, et al. Effects of Shaohuaxian capsules on lipid peroxidation of hepatocyte mitochondria [J]. Journal of Guiyang Medical College, 2004, 29(5): 406-409.
[3] Martinez FJ, Flaherty KR. Comprehensive and individualized patient care in idiopathic pulmonary fibrosis: refining approaches to diagnosis, prognosis, and treatment[J]. Chest, 2017, 151(5): 1173-1174.
[4] Li LC, Kan LD. Traditional Chinese medicine for pulmonary fibrosis therapy: progress and future prospects[J]. J Ethnopharmacol, 2017, 198: 45-63. doi: 10.1016/j.jep.2016.12.042.
[5] Suresh PS, Singh PP, Padwad YS, et al. Steroidal saponins from Trillium govanianum as α-amylase, α-glucosidase, and dipeptidyl peptidase IV inhibitory agents [J]. J Pharm Pharmacol, 2021, 73(4): 487-495.
[6] Wu AG, Teng JF, Wong VKW, et al. Novel steroidal saponin isolated from Trillium tschonoskii maxim. exhibits anti-oxidative effect via autophagy induction in cellular and Caenorhabditis elegans models [J]. Phytomedicine, 2019, 65: 153088. doi: 10.1016/j.phymed.2019.153088.
[7] Qian S, Tong S, Wu J, et al. Paris saponin VII extracted from Trillium tschonoskii induces autophagy and apoptosis in NSCLC cells [J]. J Ethnopharmacol, 2020, 248: 112304. doi: 10.1016/j.jep.2019.112304.
[8] 张忠立, 左月明, 蔡妙婷,等. 延龄草根及根茎的化学成分研究(II)[J]. 中草药, 2013, 44(20): 2808-2811. ZHANG Zhongli, ZUO Yueming, CAI Miaoting, et al. Study on chemical constituents of roots and rhizomes of Trillium tschonoskii(II)[J]. Chinese Traditional and Herbal Drugs, 2013, 44(20): 2808-2811.
[9] Yan T, Yu X, Sun X, et al. A new steroidal saponin, furotrilliumoside from Trillium tschonoskii inhibits lipopolysaccharide-induced inflammation in Raw264.7 cells by targeting PI3K/Akt, MARK and Nrf2/HO-1 pathways [J]. Fitoterapia, 2016, 115: 37-45. doi: 10.1016/j.fitote.2016.09.012.
[10] Teng JF, Qin DL, Mei QB, et al. Polyphyllin VI, a saponin from Trillium tschonoskii Maxim. induces apoptotic and autophagic cell death via the ROS triggered mTOR signaling pathway in non-small cell lung cancer [J]. Pharmacol Res, 2019, 147: 104396. doi: 10.1016/j.phrs.2019.104396.
[11] 熊秋杨, 辛光, 李世一, 等. 延龄草皂苷对小鼠急性胰腺炎和相关肺损伤的影响[J]. 华西药学杂志, 2019, 34(6): 587-591. XIONG Qiuyang, XIN Guang, LI Shiyi, et al. Effects of Trillium saponins on acute pancreatitis and related lung injury in mice [J]. West China JOurnal of Pharmaceutical Sciences, 2019, 34(6): 587-591.
[12] Yang F, Hou ZF, Zhu HY, et al. Catalpol protects against pulmonary fibrosis through inhibiting TGF-β1/Smad3 and Wnt/β-catenin signaling pathways[J]. Front Pharmacol, 2021, 11: 594139. doi: 10.3389/fphar.2020.594139.
[13] Szapiel SV, Elson NA, Fulmer JD, et al. Bleomycin-induced interstitial pulmonary disease in the nude, athymic mouse[J]. Am Rev Respir Dis, 1979, 120(4): 893-899.
[14] Ashcroft T, Simpson JM, Timbrell V. Simple method of estimating severity of pulmonary fibrosis on a numerical scale[J]. J Clin Pathol, 1988, 41(4): 467-470.
[15] Lopez AD, Avasarala S, Grewal S, et al. Differential role of the Fas/Fas ligand apoptotic pathway in inflammation and lung fibrosis associated with reovirus 1/L-induced bronchiolitis obliterans organizing pneumonia and acute respiratory distress syndrome[J]. J Immunol, 2009, 183(12): 8244-8257.
[16] Kala M, Shaikh MV, Nivsarkar M. Equilibrium between anti-oxidants and reactive oxygen species: a requisite for oocyte development and maturation[J]. Reprod Med Biol, 2016, 16(1): 28-35.
[17] He C, Larson-casey JL, Gu L, et al. Cu, Zn-superoxide dismutase-mediated redox regulation of Jumonji domain containing 3 modulates macrophage polarization and pulmonary fibrosis[J]. Am J Respir Cell Mol Biol, 2016, 55(1): 58-71.
[18] Teixeira KC, Soares FS, Rocha LGC, et al. Attenuation of bleomycin-induced lung injury and oxidative stress by N-acetylcysteine plus deferoxamine[J]. Pulm Pharmacol Ther, 2008, 21(2): 309-316.
[19] Yu WN, Sun LF, Yang H. Inhibitory effects of astragaloside IV on bleomycin-induced pulmonary fibrosis in rats via attenuation of oxidative stress and inflammation[J]. Inflammation, 2016, 39(5): 1835-1841.
[20] Park SJ, Kim TH, Lee K, et al. Kurarinone attenuates BLM-induced pulmonary fibrosis via inhibiting TGF-β signaling pathways [J]. Int J Mol Sci, 2021, 22(16): 8388.
[21] 王先丽, 任丹, 詹光杰, 等. 延龄草总皂苷对LPS诱导炎症大鼠的抗炎效果[J]. 基因组学与应用生物学, 2019, 38(8): 3697-3705. WANG Xianli, REN Dan, ZHAN Guangjie, et al. The anti-inflammatory effects of total Trillium tschonoskii Maxim Saponins on rats' inflammation induced by lipopolysaccharide [J]. Genomics and Applied Biology. 2019, 38(8): 3697-3705.
[22] 满红霞, 肖培云, 杨永寿, 等. 特发性肺纤维化的发病机制及药物治疗研究进展[J]. 中国现代应用药学, 2015, 32(8): 1024-1028. MAN Hongxia, XIAO Peiyun, YANG Yongshou, et al. Pathogenesis of idiopathic pulmonary fibrosis and its advances in cytokine treatment[J]. Chinese Journal of Modern Applied Pharmacy, 2015, 32(8):1024-1028.
[23] Fernandez IE, Eickelberg O. The impact of TGF-β on lung fibrosis: From targeting to biomarkers[J]. Proc Am Thorac Soc, 2012, 9(3): 111-116.
[24] Santibañez JF, Quintanilla M, Bernabeu C. TGF-β/TGF-β receptor system and its role in physiological and pathological conditions[J]. Clin Sci(Lond), 2011, 121(6): 233-251.
[25] Guo X, Ramirez A, Waddell DS, et al. Axin and GSK3-beta control Smad3 protein stability and modulate TGF-beta signaling[J]. Genes Dev, 2008, 22(1):106-120.
[26] Zhang M, Wang M, Tan X, et al. Smad3 prevents β-catenin degradation and facilitates β-catenin nuclear translocation in chondrocytes[J]. J Biol Chem, 2010, 285(12): 8703-8710.

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