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《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2023年05期

页码:

630-634

栏目:

论著

出版日期:

2023-10-20

文章信息/Info

Title:

Granzyme B promotes bleomycin-induced pulmonary fibrosis by activating TGF-β1/Smad3 pathway

作者:

陈向军¹; 顾 兴²; 王在强³; 王光辉¹; 王 莉¹; 方 芳¹; 金发光³; 王瑞璇²

710100 西安,西安市胸科医院重症医学二科¹
710100 西安,西安市胸科医院呼吸与危重症医学科²
710038 西安,空军军医大学第二附属医院呼吸与危重症医学科³

Author(s):

Chen Xiangjun¹;  Gu Xing²;  Wang Zaiqiang³;  Wang Guanghui¹;  Wang Li¹;  Wan Fang¹;  Jin Faguang³;  Wang Ruixuan².

¹The second Department of Intensive Care Medicine, Xi'an Chest Hospital, Xi'an 710100, China; ²Department of Respiratory and Critical Care Medicine, Xi'an Chest Hospital, Xi'an 710100, China; ³Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Air Force Medical University, Xi'an 710038, China

关键词:

肺纤维化;  颗粒酶B;  转化生长因子β₁;  Smad3

Keywords:

Pulmonary fibrosis;  Granzyme B;  TGF-β1;  Smad3

分类号:

R563

DOI:

10.3877/cma.j.issn.1674-6902.2023.05.005

摘要:

目的 分析颗粒酶B在博来霉素导致大鼠肺纤维化中的作用。方法 将21只SD大鼠随机分为三组:对照组、模型组、干预组,每组7只。模型组和干预组采用经气管内注入博来霉素(5 mg/kg)诱导大鼠肺纤维化动物模型。干预组在造模前1 h、造模第7天、第14天和第21天经尾静脉注射颗粒酶B抑制剂,其它两组经尾静脉注射等体积生理盐水,其余操作相同。28 d后处死大鼠取出肺脏,采用免疫荧光和Western blot检测颗粒酶B、胶原蛋白I、转化生长因子β₁(transforming growth factor β1)和Smad3蛋白表达情况,采用Masson染色检测肺组织纤维化情况。结果 模型组较对照组大鼠肺组织胶原蛋白Ⅰ表达上调,胶原蛋白沉积增加,肺纤维化加重。干预组较模型组胶原蛋白Ⅰ表达下调,胶原蛋白沉积减少,肺纤维化减轻。模型组较对照组大鼠肺组织TGF-β1和Smad3蛋白表达增加,干预组较模型组TGF-β1和Smad3蛋白表达降低。结论 颗粒酶B促进了博来霉素导致的大鼠肺纤维化,机制可能与激活TGF-β1/Smad3信号通路有关。

Abstract:

Objective To explore the role of granzyme B in bleomycin-induced pulmonary fibrosis in rats. Methods 21 SD rats were randomly divided into three groups: control group, model group and intervention group, with 7 rats in each group. The model group and the intervention group were injected with bleomycin(5 mg/kg)through the trachea to induce pulmonary fibrosis. The intervention group was injected with granzyme B inhibitor through caudal vein 1 hour before modeling, on day 7, 14 and 21, and the other two groups were injected with equal volume of normal saline. The remaining operations were the same. After 28 days, the rats were killed and the lungs were removed. The protein expressions of granzyme B, collagen I, TGF-β1 and Smad3 were detected by immunofluorescence and Western blot, and the pulmonary fibrosis was detected by Masson staining. Results Compared with the control group, collagen I expression, collagen deposition and pulmonary fibrosis were up-regulated in the model group. Compared with model group, collagen I expression was down-regulated, collagen deposition was reduced and pulmonary fibrosis was alleviated in the intervention group. The protein expressions of TGF-β1 and Smad3 in lung tissue of model group were increased compared with that of control group, while the protein expressions of TGF-β1 and Smad3 in intervention group were decreased compared with that of model group. Conclusion Granzyme B can promote bleomycin-induced pulmonary fibrosis in rats, and the mechanism may be related to the activation of TGF-β1/Smad3 signaling pathway.

参考文献/References:

1 Kolb P, Upagupta C, Vierhout M, et al. The importance of interventional timing in the bleomycin model of pulmonary fibrosis[J]. Eur Respir J, 2020, 55(6): 1901105.
2 Wuyts WA, Wijsenbeek M, Bondue B, et al. Idiopathic pulmonary fibrosis: Best practice in monitoring and managing a relentless fibrotic disease[J]. Respiration, 2020,99(1): 73-82.
3 于晓涛, 杨忠杰, 应真真, 等. 基于生物信息学筛选特发性肺纤维化差异基因及中药预测[J]. 中医药信息, 2022, 39(9): 68-74.
4 杨伟强, 赵 峰. 特发性肺纤维化发病机制的研究进展[J]. 医学综述, 2020, 26(9): 1684-1689.
5 Spagnolo P, Grunewald J, du Bois RM. Genetic determinants of pulmonary fibrosis: evolving concepts[J]. Lancet Respir Med, 2014, 2(5): 416-428.
6 邱 静, 孙 建, 李万成. 特发性肺纤维化中TGF-β1诱导上皮细胞间质转化作用机制研究进展[J/CD]. 中华肺部疾病杂志(电子版), 2015, 9(2): 212-214.
7 Zeglinski MR, Granville DJ. Granzymes in cardiovascular injury and disease[J]. Cell Signal, 2020, 76: 109804.
8 王在强, 金发光, 傅恩清. 颗粒酶B在组织损伤修复中的研究进展[J/CD]. 中华损伤与修复杂志(电子版), 2021,16(4): 349-352.
9 Richardson KC, Jung K, Pardo J, et al. Noncytotoxic roles of granzymes in health and disease[J]. Physiology(Bethesda), 2022, 37(6): 323-348.
10 邓玲玲, 欧阳博书, 魏 颖, 等. 上皮间质转化在特发性肺纤维化及其信号通路中的研究进展[J]. 复旦学报:医学版, 2022, 49(4): 614-619.
11 Hutchinson J, Fogarty A, Hubbard R, et al. Global incidence and mortality of idiopathic pulmonary fibrosis: a systematic review[J]. Eur Respir J, 2015, 46(3): 795-806.
12 沈忱悠, 李桂荣, 卫 栋, 等. 博来霉素诱导肺纤维化小鼠肺组织差异转录因子的筛选与功能分析[J]. 医学研究生学报, 2022, 35(8): 798-805.
13 陈叶青, 仇双逸, 范欣生, 等. 博来霉素诱导的肺纤维化形成不同时期模型大鼠血清代谢组学研究[J]. 中国药理学通报, 2022, 38(4): 512-518.
14 Shen Y, Cheng F, Sharma M, et al. Granzyme B deficiency protects against angiotensin Ⅱ-induced cardiac fibrosis[J]. Am J Pathol, 2016, 186(1): 87-100.
15 Chester D, Brown AC. The role of biophysical properties of provisional matrix proteins in wound repair[J]. Matrix Biol, 2017, 60-61: 124-140.
16 Gharaee-Kermani M, Hu B, Phan SH, et al. Recent advances in molecular targets and treatment of idiopathic pulmonary fibrosis: focus on TGF-β signaling and the myofibroblast[J]. Curr Med Chem, 2009, 16(11): 1400-1417.
17 Sriram S, Robinson P, Pi L, et al. Triple combination of siRNAs targeting TGF-β1, TGF-βR2, and CTGF enhances reduction of collagen I and smooth muscle actin in corneal fibroblasts[J]. Invest Ophthalmol Vis Sci, 2013, 54(13): 8214-8223.
18 Hu HH, Chen DQ, Wang YN, et al. New insights into TGF-β/Smad signaling in tissue fibrosis[J]. Chem Biol Interact, 2018, 292: 76-83.
19 Chen L, Yang T, Lu DW, et al. Central role of dysregulation of TGF-β/Smad in CKD progression and potential targets of its treatment[J]. Biomed Pharmacother, 2018, 101: 670-681.
20 Hu HH, Chen DQ, Wang YN, et al. New insights into TGF-β/Smad signaling in tissue fibrosis[J]. Chem Biol Interact, 2018, 292: 76-83.
21 Kang JH, Jung MY, Yin X, et al. Cell-penetrating peptides selectively targeting SMAD3 inhibit profibrotic TGF-β signaling[J]. J Clin Invest, 2017, 127(7): 2541-2554.
22 Buzza MS, Zamurs L, Sun J, et al. Extracellular matrix remodeling by human granzyme B via cleavage of vitronectin, fibronectin, and laminin[J]. J Biol Chem, 2005, 280(25): 23549-23558.
23 Choy JC, Hung VH, Hunter AL, et al. Granzyme B induces smooth muscle cell apoptosis in the absence of perforin: involvement of extracellular matrix degradation[J]. Arterioscler Thromb Vasc Biol, 2004, 24(12): 2245-2250.
24 Boivin WA, Shackleford M, Vanden HA, et al. Granzyme B cleaves decorin, biglycan and soluble betaglycan, releasing active transforming growth f factor-β1[J]. PLoS One, 2012, 7(3): e33163.
25 余洪刚, 付大海, 杨利生, 等. 复明汤调控TGF-β1/Smad3通路抑制肺成纤维细胞活化及肺上皮损伤减轻博来霉素诱导的肺纤维化[J]. 天津中医药, 2020, 37(12): 1412-1419.
26 邱 静, 李万成. TGF-β1/Smad3在博来霉素肺纤维化大鼠中的作用[J]. 成都医学院学报, 2017, 12(3): 271-276.
27 Kurschus FC, Kleinschmidt M, Fellows E, et al. Killing of target cells by redirected granzyme B in the absence of perforin[J]. FEBS Lett, 2004, 562(1-3): 87-92.
28 Cullen SP, Adrain C, Luthi AU, et al. Human and murine granzyme B exhibit divergent substrate preferences[J]. J Cell Biol, 2007, 176(4): 435-444.
29 Kaiserman D, Bird CH, Sun J, et al. The major human and mouse granzymes are structurally and functionally divergent[J]. J Cell Biol, 2006,175(4):619-630.
30 Kevin KK, Ying W, Charles S, et al. Epithelial cell alpha3beta1 integrin links beta-catenin and Smad signaling to promote myofibroblast formation and pulmonary fibrosis[J]. J Clin Invest, 2009, 119(1): 213-224.

备注/Memo

备注/Memo:

基金项目: 陕西省重点研发计划(2018ZDCXL-SF-02-03-02)
通信作者: 王瑞璇, Email: hxkwrxuan@163.com

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更新日期/Last Update: 2023-10-20