大鼠鼓室硬化建模优化及与纤维连接蛋白高表达的关联

doi:10.6040/j.issn.1671-7554.0.2020.1390

摘要/Abstract

摘要： 目的研究鼓室硬化模型构建的优化方法,探讨纤维连接蛋白在大鼠鼓室硬化模型中的表达及其在该病发病过程中的作用机制。方法 60只Sprague-Dawley大鼠随机分成3组,每组20只。A组双耳不作任何处理;B组右耳注射一次肺炎链球菌建立鼓室硬化模型,左耳不做处理;C组右耳注射两次肺炎链球菌,左耳不做处理。8周后耳显微镜观察鼓室硬化形成情况,苏木素-伊红染色(HE)观察形态学变化,免疫组化染色(IHC)确定纤维连接蛋白表达位置,Western blotting检测纤维连接蛋白的表达量。结果 8周后,A组中所有大鼠鼓膜正常,B组中35%的鼓膜出现硬化病变,C组中78.9%的鼓膜出现硬化病变;鼓室硬化大鼠的中耳黏膜可见明显炎症浸润及纤维组织增生;纤维连接蛋白的阳性表达主要位于黏膜下层;3组纤维连接蛋白的表达量分别为0.254±0.117、1.154±0.163和1.111±0.622,组间均值差异有统计学意义(F=6.768,P=0.006),两两多重比较结果显示,A1、B1和C1组间的纤维连接蛋白表达水平差异具有统计学意义(P=0.013,P=0.018)。结论鼓室内注射两次肺炎链球菌可明显增加造模成功率,纤维连接蛋白与鼓室硬化的发病机制相关。

关键词: 鼓室硬化, 动物模型, 纤维连接蛋白

Abstract: Objective To explore the optimization of tympanosclerosis model, and to investigate the expression of fibronectin and its role in the pathogenesis of tympanosclerosis. Methods A total of 60 Sprague-Dawley rats were randomly divided into 3 groups, with 20 in each group. In group A, neither ears were treated; in group B, Streptococcus pneumoniae was injected once into the right ears while the left ears were not treated; in group C, Streptococcus pneumoniae was injected twice into the right ears while the left ears were not treated. The formation of tympanosclerosis was observed with otomicroscope after 8 weeks. The morphological changes were observed with hematoxylin-eosin(HE)staining, the expression location of fibronectin was determined with immunohistochemical staining(IHC), and the expression level of fibronectin was detected with Western blotting. Results In group A, the tympanic membranes of all rats were normal, while in groups B and C, 35% and 78.9% of the tympanic membranes had tympanosclerosis. Obvious inflammatory infiltration and fibrous tissue hyperplasia could be seen in the middle ear mucosa of tympanosclerosis rats. Fibronectin was mainly located in submucosa. The expression level of fibronectin was 0.254±0.117, 1.154±0.163 and 1.111±0.622, respectively, in the three groups, with significant differences(F=6.768,P=0.006). Pairwise multiple comparison showed that there was significant difference in the expression of fibronectin among A1, B1 and C1 groups(P=0.013, P=0.018). Conclusion Two injections of Streptococcus pneumoniae can increase the success rate of modeling. Fibronectin is related to the pathogenesis of tympanosclerosis.

Key words: Tympanosclerosis, Animal model, Fibronectin

中图分类号:

R764

仇静静,吴婷,牟亚魁,崔丽梅,郭文涛,韩锋产,孙岩. 大鼠鼓室硬化建模优化及与纤维连接蛋白高表达的关联[J]. 山东大学学报 (医学版), 2021, 59(1): 22-27.

QIU Jingjing, WU Ting, MOU Yakui, CUI Limei, GUO Wentao, HAN Fengchan, SUN Yan. Optimization of rat tympanosclerosis model and the correlation between tympanosclerosis and high expression of fibronectin[J]. Journal of Shandong University (Health Sciences), 2021, 59(1): 22-27.

参考文献

[1] 仇静静, 王俊鑫, 孙岩. 鼓室硬化病因及相关因素的研究进展[J]. 中华耳科学杂志, 2020, 18(4): 792-796. QIU Jingjing, WANG Junxin, SUN Yan. Advances in research on etiology of tympanosclerosis and related factors [J]. Chinese Journal of Otology, 2020, 18(4): 792-796.
[2] Santos PF, Leal MC, Peixoto C, et al. Otomicroscopic and histologic findings of induced myringosclerosis in rats: a critical study of an experimental model [J]. Braz J Otorhinolaryngol, 2005, 71(5): 668-674.
[3] Wang Y, Ni H. Fibronectin maintains the balance between hemostasis and thrombosis [J]. Cell Mol Life Sci, 2016, 73(17): 3265-3277.
[4] Zou XZ, Liu T, Gong ZC, et al. MicroRNAs-mediated epithelial-mesenchymal transition in fibrotic diseases [J]. Eur J Pharmacol, 2017, 796: 190-206. doi: 10.1016/j.ejphar.2016.12.003.
[5] 张艳飞, 郑烨贤, 王素洁, 等. 两种鼓室硬化造模方法的比较[J]. 听力学及言语疾病杂志, 2017, 25(3): 284-287. ZHANG Yanfei, ZHENG Yexian, WANG Sujie, et al. Comparison of two methods for the modeling of tympanosclerosis in rats [J]. Journal of Audiology and Speech Pathology, 2017, 25(3): 284-287.
[6] 董晓, 赵宇. 鼓膜硬化研究现状[J]. 华西医学, 2017, 32(9): 1455-1458. DONG Xiao, ZHAO Yu. Research status of tympanic membrane sclerosis [J]. West China Medical Journal, 2017, 32(9): 1455-1458.
[7] Murphy TF, Parameswaran GI. Moraxella catarrhalis, a human respiratory tract pathogen [J]. Clin Infect Dis, 2009, 49(1): 124-131.
[8] Casey JR, Kaur R, Friedel VC, et al. Acute otitis media otopathogens during 2008 to 2010 in Rochester, New York [J]. Pediatr Infect Dis J, 2013, 32(8): 805-809.
[9] Forséni Flodin M. Macrophages and possible osteoclast differentiation in the rat bullar bone during experimental acute otitis media, with reference to tympanosclerosis [J]. Otol Neurotol, 2001, 22(6): 771-775.
[10] 李卓豪, 徐果, 张志钢. 分泌性中耳炎动物模型建立方法及研究进展[J]. 中华耳科学杂志, 2015, 13(4): 757-760. LI Zhuohao, XU Guo, ZHANG Zhigang. Establishing method and research progress of animal model of secretory otitis media [J]. Chinese Journal of Otology, 2015, 13(4): 757-760.
[11] Forséni Flodin M, Hultcrantz M. Possible inflammatory mediators in tympanosclerosis development [J]. Int J Pediatr Otorhinolaryngol, 2002, 63(2): 149-154.
[12] 柯朝阳, 杨名保, 龚桃根, 等. 大鼠急性中耳炎动物模型的建立[J]. 中华耳科学杂志, 2010, 8(3): 325-329. KE Chaoyang, YANG Mingbao, GONG Taogen, et al. A rat model of acute otitis media [J]. Chinese Journal of Otology, 2010, 8(3): 325-329.
[13] 万良财, 郭梦和, 谢南屏, 等. 鼓室硬化患者中耳黏膜及硬化灶组织病理学特点[J]. 听力学及言语疾病杂志, 2009, 17(4): 351-354. WAN Liangcai, GUO Menghe, XIE Nanping, et al. The clinicopathologic and electron-microccopic characteristics of patients with tympanosclerosis [J]. Journal of Audiology and Speech Pathology, 2009, 17(4): 351-354.
[14] Varela A, Mavroidis M, Katsimpoulas M, et al. The neuroprotective agent rasagiline mesylate attenuates cardiac remodeling after experimental myocardial infarction [J]. ESC Heart Fail, 2017, 4(3): 331-340.
[15] Lenselink EA. Role of fibronectin in normal wound healing[J]. Int Wound J, 2015, 12(3): 313-316.
[16] Yun S, Budatha M, Dahlman JE, et al. Interaction between integrin α5 and PDE4D regulates endothelial inflammatory signalling [J]. Nat Cell Biol, 2016, 18(10): 1043-1053.
[17] Hsia HC, Nair MR, Corbett SA. The fate of internalized α5 integrin is regulated by matrix-capable fibronectin [J]. J Surg Res, 2014, 191(2): 268-279.
[18] Ahn HJ, Khalmuratova R, Park SA, et al. Serial analysis of tracheal restenosis after 3D-printed scaffold implantation: recruited inflammatory cells and associated tissue changes [J]. Tissue Eng Regen Med, 2017, 14(5): 631-639.
[19] Weng CH, Li YJ, Wu HH, et al. Interleukin-17A induces renal fibrosis through the ERK and smad signaling pathways [J]. Biomed Pharmacother, 2020, 123: 109741. doi: 10.1016/j.biopha.2019.109741.
[20] Altrock E, Sens C, Wuerfel C, et al. Inhibition of fibronectin deposition improves experimental liver fibrosis [J]. J Hepatol, 2015, 62(3): 625-633.

相关文章 15

[1]	王国云,潘臧钰. 子宫腺肌病动物模型研究进展[J]. 山东大学学报 (医学版), 2022, 60(7): 48-55.
[2]	赵婷婷,齐亚娜,张颖,袁冰,韩明勇. 小鼠乳腺癌诱导转移前肺组织微环境的改变[J]. 山东大学学报 (医学版), 2022, 60(4): 24-29.
[3]	张淑莹,武晓峰,郭丽敏,乔温,彭洁琼,李大庆. 两种阿霉素心力衰竭模型及心功能进展的评估[J]. 山东大学学报 (医学版), 2020, 58(12): 1-7.
[4]	张美玲,孙卉,孙文凯,孟朝暾,邬信芳,高伟,李钦. 鼻腔滴入γ干扰素对变应性鼻炎大鼠外周血Th17/Treg细胞及相关细胞因子的影响[J]. 山东大学学报 (医学版), 2020, 58(1): 13-19.
[5]	刘聪聪,刘洪彬,符江,李雷. 多囊卵巢综合征易感基因dennd1a在斑马鱼模型中的表达特性[J]. 山东大学学报 (医学版), 2019, 57(1): 48-54.
[6]	胡玉敬,吴大勇,张文艳,边艳珠,魏强,田丛娜,常胜利. 放疗后肿瘤⁹⁹Tc^m-MNLS乏氧显像变化与乏氧诱导因子-1α表达的相关性[J]. 山东大学学报（医学版）, 2017, 55(8): 30-34.
[7]	尹妮,杨关林,姜钧文,王春田,王凤耀,贾连群,高晓宇,潘嘉祥,李芹,李佳,冯元洁,高玉竹,周鹤,张哲. 巴马小型猪冠状动脉粥样硬化模型的评价方法[J]. 山东大学学报（医学版）, 2017, 55(7): 1-5.
[8]	孟祥继，庞琦，丁锋，辛涛，杨洪安. 纹状体立体定向注射Taclo建立帕金森病大鼠模型[J]. 山东大学学报（医学版）, 2014, 52(3): 16-18.
[9]	于健, 叶瑶, 黄漓莉, 刘晓玲, 莫如芬, 杨帆, 胡璟, 周英琼, 何永玲. 巴马小型猪1型糖尿病模型胰腺病理及生化指标的变化[J]. 山东大学学报（医学版）, 2014, 52(12): 10-14.
[10]	张鹏飞, 徐晓娅, 姜曼, 毕玉莉, 许继映, 韩明勇. LPS通过PGE2-EP2信号传导通路诱导肺血管生成[J]. 山东大学学报（医学版）, 2014, 52(10): 15-19.
[11]	张斌1，陈杰2，刘雁3，张士宝2，刘双庆2，刘波2，王全颖4，杨广笑4，刘庆勇2. 分泌表达肾癌抑制融合肽的重组腺伴随病毒对肾癌移植瘤的疗效[J]. 山东大学学报(医学版), 2013, 51(5): 33-36.
[12]	徐晓娅，毕玉莉，姜曼，许继映，张鹏飞，韩明勇. 血管内皮生长因子促进肺转移相关受体通路的研究[J]. 山东大学学报(医学版), 2013, 51(11): 25-29.
[13]	闫静，侯瑾，盛颖，王波涛，祝康，康全清. 间歇性缺氧大鼠模型的建立及评价[J]. 山东大学学报(医学版), 2013, 51(06): 53-56.
[14]	刘莉1，阎明2，李强1，张海涛1. 大鼠酒精性肝损伤超微结构观察[J]. 山东大学学报(医学版), 2012, 50(8): 31-.
[15]	赵倩倩，杜其航，王守靓，王公明，张孟元. 胫骨癌痛大鼠痛觉敏化的形成及其脊髓机制[J]. 山东大学学报(医学版), 2012, 50(7): 37-.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed