美洲大蠊提取液对大鼠难愈合创面TGF-β1/Smads通路调控机制的研究

详细信息    查看全文 | 推荐本文 |

英文篇名：Study on the Regulation Mechanism of Periplaneta Americana Extract on TGF-β 1/Smads Pathway in Rats with Difficult Wound Healing 作者：曾娟妮 ; 刘筱 ; 耿越飞 ; 沈咏梅 ; 何永恒 英文作者：Zeng Juanni;Liu Xiao;Geng Yuefei;Shen Yongmei;He Yongheng;The Second Affiliated Hospital of Hunan University of Chinese Medicine;Hunan University of Chinese Medicine;Chengdu High-Tech Good Doctor First Affiliated Hospital; 关键词：美洲大蠊提取液 ; 慢性难愈合创面 ; TGF-β1 ; Smad3 ; Smad7 ; psmad3 ; psmad7 英文关键词：Periplaneta Americana extract;;chronic refractory wounds;;TGF-beta 1;;Smad3;;Smad7;;psmad3;;psmad7 中文刊名：SJKX 英文刊名：Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology 机构：湖南中医药大学第二附属医院;湖南中医药大学;成都高新好医生第一医院; 出版日期：2019-03-20 出版单位：世界科学技术-中医药现代化 年：2019 期：v.21 基金：国家自然科学基金委青年基金项目(81503585):美洲大蠊提取液对大鼠难愈合创面TGF-β1/Smads通路、ECM及DED的调控研究,负责人:曾娟妮 语种：中文; 页：SJKX201903031 页数：8 CN：03 ISSN：11-5699/R 分类号：203-210

摘要

目的:探讨美洲大蠊提取液对难愈合创面TGF-β1/Smads通路调控机制的研究。方法:选取SPF级Wistar大鼠60只,采用随机数字表法将其分为空白组(急性全层皮肤缺损组)、模型组、对照组(贝复济组)、实验组(美洲大蠊提取液组),每组15只。空白组建立全层皮肤缺损开放性创面,模型组、对照组、实验组建立全层皮肤难愈合创面。造模后3天、8天、15天、20天留取创面新鲜肉芽组织,运用免疫组化观察TGF-β1、Smad3、Smad7的表达,Western blot观察TGF-β1,Smad3、Smad7及其磷酸化的表达。结果:两种实验方法均显示空白组创面愈合速度最快,模型组愈合缓慢。免疫组化发现实验组与模型组TGF-β1、Smad7的表达比较有统计学意义(P <0.05),Smad3的表达结果显示实验组与模型组比较,3d无明显差异(P> 0.05),8天、15天、20天有统计学意义(P <0.05)。Western blot实验显示TGF-β1、Smad7、psmad3的表达,实验组与模型组比较有统计学意义(P <0.05);各组间Smad3的表达比较均无统计学意义(P> 0.05),psmad7的表达结果显示,实验组与模型组比较,3-8天无统计学意义(P> 0.05),15-20天有统计学意义(P <0.05)。结论:美洲大蠊提取液下调Smad3表达水平的现象不显著,但能引起Smad3的磷酸化;其能有效降低Smad7的表达,减少Smad7的自身磷酸化,从而减弱Smad7对TGF-β1信号转导的抑制,提高TGF-β1的表达而促进创面愈合。
        Objective: To investigate the regulatory mechanism of Periplaneta americana extract on TGF-beta 1/Smads pathway in refractory wounds. Methods: Sixty SPF Wistar rats were randomly divided into blank group(Acute fullthickness skin defect group), model group, control group(Shellfish group) and experience group(Periplaneta Americana group) with 15 rats in each group. Blank group established full-thickness skin defect open wound, model group, control group, experience group established full-thickness skin healing difficult wound. The expression of TGF-beta 1, Smad3,Smad7 and their phosphorylation were observed by Western blot on the 3 rd, 8 th, 15 th and 20 th day after the model establishment. Results: The two experimental methods showed that the wound healing rate in the blank group was the fastest and the model group healed slowly. Mmunohistochemistry showed that the expression of TGF-beta 1 and Smad7 in the experimental group was significantly different from that in the model group(P < 0.05). The expression of Smad3 showed no significant difference between the experimental group and the model group on the 3 rd day(P > 0.05). There was statistical significance on the 8 th, 15 th and 20 th day(P < 0.05). The wound healing rate in the blank group was the fastest, and the model group healed slowly. The expression levels of TGF-beta 1, Smad7 and psmad3 were significantly higher in experience group than in model group(P < 0.05). There was no significant difference in the expression of Smad3 between the groups(P > 0.05). The results of psmad7 expression showed that there was no significant difference in 3-8 days(P > 0.05) and 15-20 days(P < 0.05) between experience group and model group. Conclusion: Smad3 expression level is not significantly down-regulated by Periplaneta Americana extract, but Smad3 phosphorylation can be induced, Smad7 expression can be effectively reduced, Smad7 phosphorylation can be reduced, thus reducing Smad7 inhibition of TGF-beta 1 signal transduction, increasing the expression of TGF-beta 1 and promoting wound healing.

引文

1 Mari?lle W, Mirella G, Esther M. Altered TGF-βsignaling in fetal fibroblasts:What is known about the underlying mechanism. Wound Repair and Regeneration, 2014(22):3-13.
    2 辛宇鹏,光亚,卢一平.β转化生长因子/Smad信号通路与慢性移植肾肾病.中华移植杂志, 2009, 3(1):49-54.
    3 Lan H Y. Diverse roles of TGF-β/Smads in renal fibrosis and inflammation.Int J Biol Sci, 2011, 7(7):1056-1067.
    4 李玉兰,杨树青,龚淼,等.仙人掌联合川穹嗪外敷治疗糖尿病足42例疗效观察.齐鲁护理杂志, 2009, 15(7):35.
    5 丁宏辉.复合封闭式负压引流技术和常规VSD治疗普外科难愈性伤口对比分析.临床医药文献电子杂志, 2015, 2(13):2570-2571.
    6 伍伟明,王真权,曾娟妮,等.康复新液治疗肛管难愈合创面的临床研究.中医药导报, 2015, 21(22):71-73.
    7 曾娟妮,刘筱,伍伟明,等.美洲大蠊提取液对大鼠难愈合创面VEGF表达影响的研究.中国民族民间医药杂志, 2016, 25(19):64-66.
    8 刘筱,李文华,曾娟妮,等.美洲大蠊提取液对大鼠难愈合创面TGF-β表达的影响.中医药导报, 2018, 24(6):9-12+16.
    9 徐杰男,阙华发.补肾填精方促进大鼠慢性难愈性创面修复愈合的作用及机制探讨.四川中医, 2015(10):34-37.
    10 De Laat E H, Van den Boogaard M H, Spauwen P H, et al. Faster wound healing with topical negative pressure therapy in difficult-to-heal wounds:a prospective randomized controlled trial. Annals of plastic surgery,2011, 67(6):626-631.
    11 李莉,唐文华,徐月芳.康复新液治疗头面部烧伤36例效果观察.齐鲁护理杂志, 2010(16):57-58.
    12 魏强,杨运芳.康复新液治疗复发性阿弗他溃疡112例近期疗效观察.中国健康月刊, 2010(29):235-236.
    13 罗洪斌,韦昌南.不同治疗方法对手足外科难愈性伤口的疗效对比分析.广东微量元素科学, 2015, 22(5):54-57.
    14 Dong C. Microtubule binding to smads may regulate TGF beta activity.Mol Cell, 2000, 5:27-34.
    15 Tsukazaki T. SARA, a five domain protein that recruits smad2 to the TGF beta receptor. J Cell, 1998, 95(6):779—791.
    16 TopperJN.VascularMADs:TwonovelMAD—relatedgenesselectivelyinducible by flow in human vascular endo-thelium. Prec Natl Acad, 1997, 94:9314-9319.
    17 苏丽婷,夏时海. Smad蛋白在转化生长因子β信号转导通路中的作用与机制.医学分子生物学杂志, 2008, 5(4):352-355+360.
    18 Yang G, Yang X. Smad4-mediated TGF-beta signaling in tumorigensis.Int J Biol Sci, 2010, 6(1):18.