氧化苦参碱水凝胶通过激活角化细胞Nrf2/HO-1通路促进创面愈合  

Oxymatrine hydrogel promotes wound healing by activating Nrf2/HO-1 pathway in keratinocytes

作　　者：刘璐 刘淑丹[1] 刘晓丹 杨丽[2] 凌晨 海小明 马会明[3] 陈冬梅[1] Liu Lu;Liu Shudan;Liu Xiaodan;Yang Li;Ling Chen;Hai Xiaoming;Ma Huiming;Chen Dongmei(General Hospital of Ningxia Medical University,Key Laboratory of Stem Cell and Regenerative Medicine of Ningxia Hui Autonomous Region,Yinchuan 750004,Ningxia Hui Autonomous Region,China;School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Key Laboratory for Maintaining the Educational Ability of Medical College Students in Ningxia,Ministry of Education,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学总医院,宁夏回族自治区干细胞与再生医学重点实验室,宁夏回族自治区银川市750004 [2]宁夏医科大学基础医学院,宁夏回族自治区银川市750004 [3]宁夏医科大学,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2024年第29期4620-4627,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区项目(81960355),项目负责人:陈冬梅;宁夏自治区自然科学基金项目(2022AAC03500),项目负责人:陈冬梅;宁夏回族自治区重点研发计划项目(2022BEG03147),项目负责人:刘淑丹。

摘　　要：背景:慢性创面炎症和氧化应激阻碍了角化细胞的再生,氧化苦参碱具有抗氧化、抗炎等多种生物活性,可能具有促进创面愈合的潜在效应。目的:探讨氧化苦参碱对创面愈合的作用,以及对H_(2)O_(2)诱导人角质形成细胞系HaCaT细胞氧化应激损伤的保护作用。方法:①体内实验:分别制备含0,0.05,0.1,0.2 g/L氧化苦参碱的甲基丙烯酰化透明质酸(HAMA)水凝胶。在75只糖尿病小鼠背中部制作直径12 mm的全层皮肤缺损模型,随机分5组干预,每组15只:模型组创面包扎固定,单纯水凝胶组以HAMA水凝胶覆盖创面,低、中、高剂量氧化苦参碱组分别以含0.05,0.1,0.2 g/L氧化苦参碱的HAMA水凝胶覆盖创面,光固化后包扎固定,14 d内进行相关指标的检测。②体外实验:将人角质形成细胞系HaCaT分5组培养,正常组常规培养,H_(2)O_(2)组及低、中、高浓度氧化苦参碱组均加入H_(2)O_(2)干预4 h,然后分别更换为含0,0.05,0.1,0.2 g/L氧化苦参碱的培养基,培养24 h后进行相关指标的检测。结果与结论:①体内实验:与模型组比较,单纯水凝胶组小鼠创面愈合率无明显变化,低、中、高剂量氧化苦参碱组治疗后7,14 d的创面愈合率升高(P<0.05)。治疗后14 d的创面样本切片病理观察显示,与模型组相比,中、高剂量氧化苦参碱组再生表皮层厚度、显微血管数量与胶原沉积均增加(P<0.05)。术后7 d的创面样本Western blotting检测显示,与模型组相比,中、高剂量氧化苦参碱组肿瘤坏死因子α与白细胞介素6的蛋白表达降低(P<0.05)。②体外实验:CCK-8检测、EdU及Ki67染色显示,与H_(2)O_(2)组相比,中、高浓度氧化苦参碱组细胞增殖能力明显升高(P<0.05)。与H_(2)O_(2)组相比,中、高浓度氧化苦参碱组线粒体膜电位升高(P<0.05)、活性氧含量降低(P<0.05)。Western blotting检测显示,与H_(2)O_(2)组相比,高浓度氧化苦参碱组Nrf2核蛋白、Nrf2总蛋白、HO-1蛋白及超氧化BACKGROUND:Inflammation and oxidative stress contribute to the barriers of regeneration in chronic wound.Oxymatrine has various biological activities,such as anti-oxidation,anti-inflammation and so on,which may have the potential effect of promoting wound healing.OBJECTIVE:To investigate the effect of oxymatrine on wound healing and the protective effect on H_(2)O_(2)-induced oxidative stress injury in human keratinoid cell line HaCaT cells.METHODS:(1)In vivo experiment:Hyaluronic acid methacryloyl hydrogels containing 0,0.05,0.1,0.2 g/L oxymatrine were prepared.A full-layer skin defect model with a diameter of 12 mm was made in the back of 75 diabetic mice and randomly divided into five groups for intervention,with 15 mice in each group.The wounds of the model group were bandaged and fixed.The wounds of the hydrogel group were covered with hyaluronic acid methacryloyl hydrogel.The wounds of the low-dose,moderate-dose and high-dose oxymatrine groups were covered with hyaluronic acid methacryloyl hydrogel containing 0.05,0.1,and 0.2 g/L oxymatrine,respectively,and then bandaged and fixed after light curing.Relevant indicators were detected within 14 days.(2)In vitro experiment:Human keratinocyte line HaCaT was divided into five groups.The normal group was cultured conventionally.H_(2)O_(2)group and low-,moderate-and highconcentration oxymatrine groups were treated with H_(2)O_(2)for 4 hours,and then the medium was replaced with medium containing 0,0.05,0.1,and 0.2 g/L oxymatrine,respectively,and the relevant indexes were detected after 24 hours of culture.RESULTS AND CONCLUSION:(1)In vivo experiment:Compared with the model group,the wound healing rate of mice in the hydrogel group had no significant change.The wound healing rate of mice in the low-,moderate-and high-dose oxymatrine group was increased at 7 and 14 days after treatment(P<0.05).Pathological observation of wound section 14 days after treatment showed that compared with the model group,the thickness of regenerated epidermal layer,the number of microves

关 键 词：氧化苦参碱 水凝胶 抗氧化 创面愈合 Nrf2/HO-1信号通路 HaCat细胞 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R-331