CN115177744A - 一种促进RISI愈合的miR181A@EM-Hb-IFI6材料及其制备方法和应用 - Google Patents
一种促进RISI愈合的miR181A@EM-Hb-IFI6材料及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种促进RISI愈合的miR181A@EM‑Hb‑IFI6材料及其制备方法和应用。将DSPE‑PEG‑NHS溶于PBS溶液中，加入miR 181a‑NH2，室温培养过夜；然后进行透析、纯化得到DSPE‑PEG‑miR‑181a；将含有血红蛋白的红细胞膜、IF16与步骤(1)得到的DSPE‑PEG‑miR‑181a混合，超声处理后过滤，再经过透析得到miR181A@EM‑Hb‑IFI6材料。本发明首次将miR‑181a用于创面愈合材料，制备得到的miR181A@EM‑Hb‑IFI6可促进创面愈合，减少活性氧的表达，通过启动SSBP1/HSF1信号通路促进RISI愈合，还可改善免疫微环境。
Description
技术领域
本发明涉及创面修复材料技术领域，具体涉及一种促进RISI愈合的 miR181A@EM-Hb-IFI6材料及其制备方法和应用。
背景技术
放射治疗已成为恶性肿瘤的三大治疗手段之一，很多地区的恶性肿瘤患者接受放射治疗，显示了放射肿瘤学在肿瘤治疗中的重要作用和地位。放射性皮肤损伤(RISI)是放射治疗的主要不良反应和剂量限制因素，目前尚无有效的治疗方法。然而，皮肤辐射反应是常见的不良反应，限制了放疗剂量和治疗效果。在接受放射治疗的癌症患者中，高达95％的患者出现皮肤反应，近10％的患者出现严重皮损，放射性皮肤损伤(RISI)的确切机制尚不清楚，缺乏规范和统一的RISI防治。
α-干扰素诱导蛋白6(IFI6)是一类Ⅰ型干扰素刺激基因，调节细胞凋亡和免疫应答。 CTD-3252C9。可通过结合IRF1和抑制IFI6的转录，促进细胞凋亡和抑制细胞生长，成为胰腺癌潜在的治疗靶点。IFI6诱导登革热和乙型肝炎病毒感染，减少血管内皮细胞和乙型肝炎病毒特异性CD8+T细胞凋亡，发挥抗病毒功能。激活转录因子3(ATF3)下调其新靶点IFI6和IFI27，抑制舌鳞癌细胞的生长和迁移。IFI6在角质形成细胞中的过度表达减少了其自发凋亡，解释了皮肤银屑病的发生和发展。这些研究证实了IFI6在不同条件下的诱导表达及其抗凋亡功能。然而，正如其他报道的那样，IFI6也可以被激活并导致凋亡。最新的发现表明，Affymetrix人HTA2.0微阵列可以确定miRNA表达谱。miRNA表达谱显示13个基因下调，50个基因上调，IFI6上调。IFI6的过度表达促进细胞增殖，减少细胞凋亡和活性氧(ROS)的产生；相反，这种过度表达增加了HaCaT和人皮肤成纤维细胞 (WS1)的放射敏感性。
近年来的研究表明，非编码RNA在调节肿瘤的发生和发展中起着重要的作用。miR181A-5p通过与CCAT结合，在体内抑制结肠癌肿瘤生长，促进细胞凋亡。因此， miR181A是一种肿瘤抑制因子。ANRIL表达miR181A并促进喉癌增殖和上皮间质转化。研究表明缺氧可延缓RISI的愈合。大量的研究致力于创造半合成或全合成的人工氧(O2) 载体。仿生纳米药物缓释系统是药物缓释系统的研究热点之一。该体系以其良好的生物相容性和低免疫原性，解决了传统纳米药物代谢快、半衰期短的问题。常用的细胞膜仿生纳米载体包括EM、干细胞膜和肿瘤细胞膜。血红蛋白纳米颗粒含有疏水结构域和许多药物结合位点。该材料具有生物相容性好、免疫原性低、易代谢等优点，易于制备，具有良好的载药能力。它已广泛应用于各种药物的递送。红细胞的胞内区域具有能与血红蛋白结合的位点，为EM实现血红蛋白纳米颗粒的定向包裹提供了思路。Hb是特异性靶向TAMs 的合适载体。作为红细胞的携氧成分，Hb通过原位释放O2来缓解肿瘤组织中的缺氧。因此针对放射性皮肤损伤的愈合，需要一种含有红细胞膜的创面修复材料，对HaCaT细胞无毒性，能促进其迁移、血管化、抑制凋亡使其具有体内外协同抗辐射能力；促进创面愈合，减少活性氧的表达，通过启动SSBP1/HSF1信号通路促进RISI愈合，还可改善免疫微环境。
发明内容
针对上述现有技术，本发明的目的是提供一种促进RISI愈合的 miR181A@EM-Hb-IFI6材料及其制备方法和应用。本发明首次将miR-181a用于创面愈合材料，制备得到的miR181A@EM-Hb-IFI6可促进创面愈合，减少活性氧的表达，通过启动SSBP1/HSF1信号通路促进RISI愈合，还可改善免疫微环境。
为实现上述目的，本发明采用如下技术方案：
本发明的第一方面，提供miR181A@EM-Hb-IFI6材料的制备方法，包括以下步骤：
(1)将DSPE-PEG-NHS溶于PBS溶液中，加入miR 181a-NH2，室温培养过夜；然后进行透析得到DSPE-PEG-miR-181a；
(2)将含有血红蛋白(Hb)的红细胞膜(EM)、IF16与步骤(1)得到的 DSPE-PEG-miR-181a混合，超声处理后过滤，再经过透析得到miR181A@EM-Hb-IFI6材料。
优选的，步骤(1)，所述PBS溶液为pH＝8的无菌PBS溶液，所述PBS溶液的浓度为0.01M；DSPE-PEG-NHS溶于PBS溶液后，浓度为0.2M。
优选的，步骤(1)，所述DSPE-PEG-NHS与miR 181a-NH2的质量比为1：1。
优选的，步骤(1)，透析所用膜为孔径50nm的聚碳酸酯膜，透析时间为6小时/次，透析2次。
优选的，步骤(2)，所述含有血红蛋白的红细胞膜、IF16与DSPE-PEG-miR-181a 的质量比为1：1：1。
优选的，步骤(2)，所述超声的功率为200W、频率为28KHz，超声的温度为25℃，超声的时间为1h。
优选的，步骤(2)，所述过滤为过滤膜为200nm的脂质体挤出机。
优选的，步骤(2)，所述透析为使用孔径50nm的聚碳酸酯膜进行透析。
本发明的第二方面，提供上述制备方法制备得到的miR181A@EM-Hb-IFI6材料。
本发明的第三方面，提供miR181A@EM-Hb-IFI6材料在促进RISI愈合中的应用。
本发明的有益效果：
(1)本发明通过设计将红细胞膜(EM)和血红蛋白(HB)修饰IFI6和miR181a结合在一起得到一种促进RISI愈合的材料，以血红蛋白/IFI6蛋白包被红细胞膜和miR181a外膜的纳米仿生给药系统，增强局部循环和生物相容性，发挥长期稳定的作用。
(2)本发明在通过细胞学研究表明，miR181a@em-hb-ifi6对HaCaT细胞无毒性，能促进其迁移、血管化、抑制凋亡，并能在照射后表达IFI6；促进创面愈合，减少活性氧的表达。还能通过启动SSBP1/HSF1信号通路促进RISI愈合，可改善免疫微环境。
附图说明
图1：miR181a@em-hb-ifi6的制备及其在放射性皮肤损伤愈合中的应用示意图。
图2：MiR181A@Em-Hb-IFI6的表征。(A)miR181a@em-hb-ifi6的透射电镜图像。(B)miR181a@em-Hb和miR181a@em-Hb-IFI6的电泳图谱。(C)粒度和电位分析。D＝147nm， PDI＝0.19，zeta电位＝-10.08mV。(D)MiR181A@EM-Hb-IFI6的紫外光谱。 (E)MIR181A@EM-Hb-IFI6的FTIR。(F)MIR181A@EM-HB-IFI6的XPS。 (G)MIR181A@EM-Hb-IFI6的EDS。测试元素包括S、P、N、Fe、C(H)相对miR181a表达的miR181a和miR181a@em-Hb-IFI6。
图3：miR181a@em-hb-ifi6的体外细胞学研究。(A)HaCaT细胞在miR181a@em-hb-ifi6 上培养7天后的FITC/DAPI染色图像。(B)IFI6-PDA@GO/SA对G+(MRSA)和G-(大肠杆菌)细菌的抗菌活性。(C)在miR181a@em-hb-ifi6上培养的HaCaT细胞第7天进行CCK-8 测定。(D)细菌计数*P<0.05。(E)大肠杆菌数*P<0.05。
图4：miR181a@em-hb-ifi6的体外细胞学研究。(A+D)流式细胞术及其总凋亡率结果。 IBI6蛋白印迹(B+E)。(C)细胞克隆形成试验。*P<0.05。
图5：miR181a@EM-HB-IFI6的体外细胞学研究。(A)使用miR181a@EM-HB-IFI6进行HaCaT细胞划伤迁移。(B)试管形成实验。(C)a的迁移面积百分比(D-E)b的节点数和管总长度*P<0.05。
图6：miR181a@EM-HB-IFI6的体内小鼠研究。(A)医用电子直线加速器及其在小鼠RISI模型中的运行过程。(B)第1、14天伤口照片和第14天HE染色照片。(C)各组第14 天创面面积。(D)完全愈合时间。(E)第14天RISI评级。(F-G)第14天创面微血管密度及肉芽组织厚度。*P<0.05。
具体实施方式
应该指出，以下详细说明都是例示性的，旨在对本申请提供进一步的说明。除非另有指明，本文使用的所有技术和科学术语具有与本申请所属技术领域的普通技术人员通常理解的相同含义。
正如背景技术部分介绍的，IFI6在照射后的皮肤细胞内高表达，并且显著增加皮肤细胞的辐射抗性，具体表现为促进照后皮肤细胞的增殖能力，降低辐射诱导的细胞凋亡水平。与此同时，IFI6在照后的皮肤细胞内向细胞核聚集，与另一入核蛋白SSBP1发生相互作用，显著影响内质网应激通路中热休克调节因子1(HSF1)的转录活性。所以，IFI6可以通过调控HSF1介导的内质网应激过程从而抑制辐射诱导的皮肤细胞损伤，降低皮肤细胞的辐射敏感性。但如何将IFI6具体应用于促进RISI愈合，还未有相关报道，因为促进RISI 愈合还需要克服生物相容性、粘附性、抗氧化性等多个问题。
基于此，本发明的目的是提供一种促进RISI愈合的miR181A@EM-Hb-IFI6材料及其制备方法和应用。本发明首次将miR-181a用作RISI创面愈合材料，设计一种以血红蛋白 /IFI6蛋白包被红细胞膜和miR181a外膜的纳米仿生给药系统，研究其对HaCaT细胞的作用，以及对RISI创面愈合的影响。本发明可为生物材料治疗RISI提供依据。通过 miR181a@em-hb和IFI6之间的协同作用，在介导活性氧诱导的癌细胞辐射敏感性中发挥作用，促进进RISI愈合。
为了使得本领域技术人员能够更加清楚地了解本申请的技术方案，以下将结合具体的实施例详细说明本申请的技术方案。
本发明实施例中所用的试验材料均为本领域常规的试验材料，均可通过商业渠道购买得到。
说明：陆军医科大学实验动物系提供了体重25克的雄性BALB/c小鼠，在典型环境下，在相对湿度为50％和环境温度为25℃的塑料笼中按12h昼夜节律单独喂养。动物实验由军医大学动物伦理委员会批准，所有操作均按相关伦理规范进行。
IFI6购自SAB公司(美国马里兰州)。
EM购自中国西安瑞希材料技术公司。
抗IFI6和抗磷HSF1购自中国北京Bioss公司。
抗SSBP1购自FineTest公司(中国武汉)。
miR181a-NH2购自吉玛基因股份有限公司。
miR181a中的有义链:Aacauucaacgcugucgugugagu(SEQ ID NO.1)，反义链:Ucccgacagcguugaauguuuu(SEQ ID NO.2)。
实施例1
MiR181A@EM-HB-IFI6的制备过程如图1所示。
(1)从小鼠红细胞中提取EM(保留血红蛋白)并定量。
(2)制备DSPE-PEG-miRNA-181a:将适量DSPE-PEG-NHS溶于无菌PBS(pH＝8)中，加入适量mir181a-NH2，室温培养过夜；然后对DSPE-PEG-miRNA-181a进行透析，纯化并去除未反应的DSPE-PEG-NHS和miRNA-181a-NH2。
(3)将适量红细胞膜与一定量的DSPE-PEG-miRNA-181a和IF16混合，用水超声和脂质体挤出机(200nm滤膜)处理，用纳米透析装置(孔径50nm的聚碳酸酯膜)透析，去除卸载的DSPE-PEG-miRNA-181a和IF16。最后，将无菌水加入体积至2毫升。
(4)产品合成过程中使用的仪器、消耗品和水已用DEPC水消毒。操作在无菌条件下进行，涉及mir181a的操作在冰上进行。
用动态光散射法测定了纳米粒子的粒径和zeta电位，用透射电镜观察了EM的形貌，证实了miR181a与EM的成功连接。用Nicolet 6700FTIR光谱仪(4000-600cm-1)记录傅里叶变换红外(FTIR)光谱。紫外可见近红外光谱测量材料的紫外光谱仪/近红外光吸收效应。能量色散谱(EDS)/X射线光电子能谱(XPS)分析了材料的元素组成。用Western印迹法测定材料的蛋白质组成。用共聚焦激光扫描显微镜对HaCaT细胞进行了表征。
扫描电镜图像显示miR181a@em-hb-ifi6的纳米级亚球形形貌(图2a)。 miR181A@Em-Hb-IFI6具有典型的核壳结构，直径约为50～100nm。miR181A@em-Hb-IFI6 的平均粒径为147nm，正zeta电位为-10.08mV，多分散指数(PDI)＝0.19(图2C)。这些结果也表明EM在HB/IFI6表面的成功包覆，一层EM的厚度为7.8nm。
为了进一步测试EM膜和HB/IFI6包覆情况，进行了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析(图2B)。发现miR181A@em-hb-ifi6表达HB/IFI6，与现有研究中的HB和IFI6蛋白条带一致，表明EM和HB/IFI6的检测成功。
为了进一步验证miR181a是否包覆在EM-HB-IFI6上，用TaqMan qPCR分析miR181a在miR181a和miR181a@EM-HB-IFI6中的表达。结果(图2h)表明miR181a成功地负载在EM-HB-IFI6上，载药率可达81％左右，SDS-PAGE显示Hb-PCL16000偶联物在16 和32kDa处有明显的条带，而天然Hb仅在32kDa处有明显的条带。紫外光谱显示该材料的吸收峰约为400nm(图2d)。FTIR显示了两个特征吸收峰，一个在1640cm-1处，另一个在3300cm-1处(图2E)。目前尚无IFI6特征吸收峰的报道；但3300cm-1位置可能与IFI6 的组成密切相关。
FTIR元素映射图表明，S、P、N、Fe、C元素在MIR181A@EM-HB-IFI6 NCs中分布均匀(图2G)。miR181A@EM-Hb-IFI6 NCs的X射线光电子能谱分析进一步证实了S、P、 N、Fe和C的存在(图2F)。Fe的分布也代表了Hb在材料中的分布，表明Hb成功地负载在材料中。
试验例
1.试验和材料
1.1.生物相容性评价及细胞形态学观察
用CCK-8测定细胞毒性。细胞接种前，样品在96孔板中紫外线消毒60min/侧，并在Dulbecco改良Eagle培养基(DMEM)中预孵育。将HaCaT细胞以2×103细胞/孔的密度接种到A-E组，加入5％胎牛血清和1％青霉素/链霉素的DMEM中，温度37℃。培养1，4， 7天后，用CCK-8测定细胞活力。
为了研究HaCaT对细胞骨架形态的影响，将HaCaT接种在A-E组培养基上24h。然后用预温的磷酸盐缓冲盐水洗涤细胞三次，并在室温下用预温的4％多聚甲醛固定细胞。我们用0.5％Triton X-100磷酸盐缓冲盐水冲洗固定细胞，浸泡5分钟。然后将细胞与制备好的酚氧化酶在37℃下与DAPI(2-(4-脒基苯基)-6-吲哚氨基甲酸二盐酸盐)孵育30分钟，孵育5分钟。用共聚焦激光扫描显微镜(德国蔡司780公司)获得染色细胞的荧光图像，然后用DAPI和鬼笔环肽对HaCaT进行染色，然后进行荧光观察。
1.2.细菌共培养
MRSA(耐甲氧西林金黄色葡萄球菌)和大肠杆菌(大肠杆菌)来自陆军医科大学。将细菌扩增(培养过夜)至1×109CFU/ml，用Luria-Bertani培养基稀释至1×104CFU/ml。抽出等量的菌液涂在盘子上。37℃培养24h后，测定细菌数量的变化。
1.3.划伤迁移
HaCaT在完全培养基中汇合生长，在24个孔板(2×104/孔)中播种。用移液管、针尖和蔡司视频显微镜对单层进行24小时检查(0h)。对于每组5个重复的单个实验，使用ImageJ1.48V软件(NIH，美国)进行具体测量；试验一式三份。
1.4.体外试管形成试验
混合2×104个HaCaT细胞接种到Matrigel涂层的96孔板上，然后加入A-E组。刺激24h后，用倒置相位显微镜(Olympus，美国)拍摄新形成的管。每组有五个重复。每个实验进行三次。
1.5.流式细胞术
细胞凋亡检测采用流式细胞术和Annexin V-FITC/PI凋亡检测试剂盒(DojindoMolecular Technologies，Japan)进行。
收集细胞并将其悬浮在含有5μL Annexin V-FITC和5μL PI的200μL结合缓冲液中。应用流式细胞仪检测RISI创面免疫微环境中的CD4+、CD8+、NK细胞和M1细胞，并用美国Beckman Coulter公司的荧光激活细胞分选仪(FACS)软件对细胞进行分析。
1.6.细胞克隆形成试验
方法参照1.1；每孔用0.005％结晶紫1ml染色1h，显像后用Image Pro Plus6.0计算群落数。
1.7.RISI小鼠模型
每只小鼠腹腔注射戊巴比妥约0.2ml麻醉。该直线加速器发射6Mev电子射线(一次照射30Gy，照射场1cm×1cm，剂量300cGy/min，10min)。源皮肤距离为1米，剩余皮肤用铅板封堵。每组照射后，用材料覆盖照射部位，每隔一天更换一次，共7～14d，分为5组，每组5只。A组仅为去毛小鼠，B组仅为放疗，C组为放疗+miR181a@em-Hb， D组为放疗+IFI6，D组为放疗+miR181a@em-Hb-IFI6。
采用IPP6.0软件，比较愈合前后创面面积，计算愈合率。
创面愈合率＝(初始创面面积-特定愈合时间后的创面面积)/初始创面面积×100％， RISI评分采用Douglas和Fowler评分法。
1.8.苏木精-伊红(HE)染色及组织学分析
每只小鼠伤后14天取创面标本制作石蜡切片进行HE染色。选择高质量图像进行HE染色，用盲法测量了新生上皮的长度。
1.9.蛋白印迹和免疫组化染色检测SBPP1/HSF1的表达
伤后14天，取10mm×10mm的小方块，包括新生表皮和肉芽组织，立即在液氮中冷冻。严格参照制造商的说明(Varioskan Flash；Thermal Scientificient，美国)，采用双线粒体酸法测定上清液的蛋白质浓度。抗IFI6(Bioss，中国)、抗SBPP1(FineTest，中国) 和抗HSF1(Bioss，中国)抗体在1:1000稀释。辣根过氧化物酶标记山羊抗兔二抗(中国中山生物公司)稀释至1:2000。收集PVDF(聚偏氟乙烯)膜并送去化学发光观察(美国热科学)。
伤口组织切片分离复水，热介导抗原在柠檬酸钠缓冲液中95℃孵育回收。最后用光学显微镜(德国徕卡，ctr6000)获得图像。
1.10.实时定量PCR和活性氧测定
用7500实时PCR系统(Application Biosystems Instruments)和SYBR GreenMaster Mix(TOYOBO，QPK-201)对NLRP3和ROS进行实时PCR。用酶联免疫吸附试剂盒测定各组ROS变化。
1.11.统计分析
每个实验一式三份，所有数据以均值+均值的标准差表示。两组比较采用学生t检验，差异有统计学意义，P<0.05。
2.结果和讨论
2.1 miR181a@em-Hb-IFI6的生物相容性和抗菌活性
体外细胞学研究分为5组：A组为正常HaCaT细胞；B组为HaCaT细胞+4Gy照射； C组为HaCaT细胞+4Gy照射+miR181a@em-Hb；D组为HaCaT细胞+4Gy照射+IFI6；E 组为HaCaT细胞+4Gy照射+miR181a@em-Hb-IFI6。
如图3a所示，HaCaT细胞与miR181a@em-hb-ifi6共培养7天，FITC/DAPI染色显示细胞形态无明显差异；细胞核和细胞质完整。CCK-8法显示，4-Gy照射后第4天，B组和E组HaCaT细胞的OD值均降低，提示照射明显抑制细胞生长(图3B)。第7天E组 OD值明显恢复，且显著高于B组(P<0.05)，提示miR181a@em-hb-ifi6在短期(1-4d)内可能不影响照射细胞的生长，而在长期(>5d)照射细胞的生长曲线恢复正常。
皮肤创面细菌感染延缓创面愈合，甚至引起创面恶化。提高抗菌性能是开发新型创面敷料的必要条件。如图3B、D、E所示，对照组MRSA(G+)和大肠杆菌(G-)生长良好。 EM-HB和EM-HB-IFI6均不能有效抑制细菌的生长(P>0.05)，而miR181a@EM-HB-IFI6 表现出抑菌活性(P<0.05)。miR181a通过与结肠癌中的CCAT结合，在体内抑制肿瘤生长，促进结肠癌细胞凋亡。因此，miR-181a可能通过炎症途径抑制细菌生长。
2.2miR181a@em-hb-ifi6的体外细胞学研究
用流式细胞术进一步分析IFI6在细胞凋亡中的潜在作用。放疗显著提高细胞凋亡率 (图4a+4d)。E组细胞凋亡率低于B组(P<0.05)，E组的作用高于D组。说明 miR181a@em-hb-IFI6能显著降低细胞凋亡率。
免疫印迹分析中IFI6的表达如图4B+4E所示。E组IFI6的表达明显高于B组(P<0.05) 和D组(P<0.05)，说明该物质可能通过内吞作用进入HaCaT细胞胞浆，从而发挥相关作用。
为了验证IFI6蛋白对HaCaT细胞的辐射保护作用，我们采用了“金标准”范式。细胞克隆形成实验表明，4-Gy照射后，单纯照射组(B组)的相对克隆数比照射前减少了 45％(P<0.05)。相比之下，E组的相对克隆数仅下降约20％(P<0.05)(图4c)。说明IFI6蛋白和miR181a@em-hb可以协同增强细胞的抗辐射能力，有利于细胞克隆的形成。
2.3miR181a@em-hb-ifi6的体外细胞学研究
如图5A+4C所示，将HaCaT细胞与各组中的材料共培养约24小时，然后进行细胞划痕试验。B组24h迁移率明显降低(P<0.05)，可能与辐射对细胞迁移的影响有关，E组的迁移抑制作用明显改善，提示该材料能提高照射后细胞的迁移率(P<0.05)。值得注意的是，E组细胞迁移率高于D组(P<0.05)，说明IFI6蛋白表达增加促进了HaCaT细胞的迁移。本发明发现Ifi6在HaCaT细胞中的过度表达通过抗凋亡促进细胞存活，并通过促进细胞迁移引导存活细胞的局部聚集，说明Ifi6和miR181a@em-hb可以共同促进肿瘤细胞存活。
如图5B+5D+5E所示，将血管内皮细胞与不同组材料共培养后，进行Matrigel升血管生成实验约24h。A组正常细胞的血管数目最多，血管长度最长。B组4-Gy照射后血管形成明显受限，D组前体材料部分恢复血管形成(P<0.05)；E组为miR181a@em-hb-IFI6材料。 5-羟色胺的促血管生成作用以E组最好(P<0.05)。
2.4 miR181a@EM-HB-IFI6对RISI小鼠模型的影响
体内细胞学研究分为5组：A组为正常小鼠；B组为小鼠+30Gy照射；C组为小鼠+30Gy辐照+miR181a@EM-HB；D组为小鼠+30Gy照射+IFI6；E组为小鼠+30Gy照射 +miR181a@EM-HB-IFI6。
在图6A中，左图为电子束放射治疗设备，右图为小鼠动物模型的定位和建模过程。如图6B所示，所有小鼠在第1天剃毛后成像，建立模型。第14天，再次给小鼠拍照。处死小鼠，收集局部皮肤组织进行HE染色(图6C)。与B组相比，D组和E组促进创面愈合 (p<0.05)，E组显著高于D组(p<0.05)，说明IFI6促进表皮细胞迁移和增殖，从而促进创面愈合。如图6A所示，各组第14天均有明显的炎症反应。e组表皮逐渐形成，部分基因药物对RISI有潜在的治疗作用。基于基因的药物必须在局部保持足够的浓度，以维持药物的持续释放，同时保持活性。具有足够载药能力的纳米材料可以解决这些问题。在本发明中，我们增强了Hb和IFI6结合的Hb/IFI6对EM NPs的持续疗效，形成纳米复合物 miR181a@EM-HB-IFI6，该复合物是在x射线照射前皮下注射到皮肤获得的。显著提高了 Hb/IFI6的稳定性，并稳定、激活和释放了来自miR181a@EM-HB-IFI6的miR181a、Hb 和IFI6。
miR181a@em-hb-ifi6纳米系统具有以下优点：
i)em包裹的局部注射材料具有良好的生物相容性，有利于材料在局部血管中的循环。 HB携带氧气，缓解伤口缺氧。因此，将HB/IFI6包封后，有利于HB/IFI6渗透到角质层和输送到皮肤，有力地改善了HB/IFI6全身吸收差、生物利用度低、注射给药时全身大量清除的缺陷；ii)IFI6促进RISI创面细胞增殖和迁移；iii)本发明的纳米系统协同 miRNA181a和IFI6抗RISI的抗氧化和抗衰老作用。本发明的miR181a@EM-HB-IFI6制剂具有良好的自由基清除能力，细胞毒性小，可降低HaCaT细胞的脂质过氧化、细胞内 ROS和基质金属蛋白酶的表达。说明本发明的材料在防御RISI方面的潜在应用。如图6d 所示，从总愈合时间来看，单纯照射组(B组)小鼠完全愈合时间约为44天。 miR181a@EM-HB显著缩短愈合时间至30天(p<0.05)。E组愈合效果最强，完全愈合时间约为24D，显著优于D组(p<0.05)。
放射治疗肿瘤组(RTOG)评分常用来评价RISI小鼠。本研究采用RTOG评分对创面14天愈合情况进行主观评价。B组和C组评分基本相同，创面RTOG 3-4级，除皮肤皱褶外，主要为融合、湿脱/坑状水肿，部分创面甚至出现溃疡、出血、坏死(图6E)。D组创面明显改善，RTOG 2～3，呈斑片状湿脱/中度水肿。E组创伤主观程度最好(RTOG 2级)，主要表现为鲜亮红斑。
在创面修复过程中，血管形成的数量和质量直接影响创面愈合程度。HE染色结果分析(图6B)，肉芽组织厚度(图6F)与伤口血管密度(图6G)基本一致。辐射降低肉芽组织厚度和血管密度，导致伤口。新的上皮细胞生长和伤口血管化过程被抑制，延迟伤口愈合。miR181a@EM-HB显著增加创面肉芽组织厚度，增加创面血管密度，促进创面愈合。
2.5结论
本发明设计并制备了一种结合miR181a和IFI6的用于RISI创面愈合的纳米材料。miR181a@EM-HB-IFI6通过Hb携氧增加创面组织中氧含量，引发HIF-1α的有效抑制。 EM在纳米材料中具有良好的生物相容性和循环性。氧的增加和HIF-1α的抑制能清除RISI 创面组织中的ROS。miR181a具有良好的抗电离辐射性能，IFI6通过激活SSBP1/HSF1信号通路协同降低氧化应激和炎症反应。
miR181a@EM-HB-IFI6改善RISI伤口炎症，诱导肉芽组织形成，血管生成和胶原蛋白沉积，导致更快的伤口闭合。此外，miR181a@EM-HB-IFI6还可以改善免疫微环境，为 RISI伤口修复提供了一个有价值的选择和有前途的策略。
以上所述仅为本申请的优选实施例而已，并不用于限制本申请，对于本领域的技术人员来说，本申请可以有各种更改和变化。凡在本申请的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本申请的保护范围之内。
SEQUENCE LISTING
<110> 中国人民解放军西部战区总医院
<120> 一种促进RISI愈合的miR181A@EM-Hb-IFI6材料及其制备方法和应用
<130> 2022
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 24
<212> DNA
<213> miR181a
<400> 1
aacauucaac gcugucgugu gagu 24
<210> 2
<211> 22
<212> DNA
<213> miR181a
<400> 2
ucccgacagc guugaauguu uu 22
Claims (10)
1.miR181A@EM-Hb-IFI6材料的制备方法，其特征在于，包括以下步骤：
(1)将DSPE-PEG-NHS溶于PBS溶液中，加入miR 181a-NH2，室温培养过夜；然后进行透析得到DSPE-PEG-miR-181a；
(2)将含有血红蛋白的红细胞膜、IF16与步骤(1)得到的DSPE-PEG-miR-181a混合，超声处理后过滤，再经过透析得到miR181A@EM-Hb-IFI6材料。
2.根据权利要求1所述的制备方法，其特征在于，步骤(1)，所述PBS溶液为pH＝8的无菌PBS溶液，所述PBS溶液的浓度为0.01M；；DSPE-PEG-NHS溶于PBS溶液后，浓度为0.2M。
3.根据权利要求1所述的制备方法，其特征在于，步骤(1)，所述DSPE-PEG-NHS与miR181a-NH2的质量比为1：1。
4.根据权利要求1所述的制备方法，其特征在于，步骤(1)，透析所用膜为孔径50nm的聚碳酸酯膜，透析时间为6h/次，透析2次。
5.根据权利要求1所述的制备方法，其特征在于，步骤(2)，所述含有血红蛋白的红细胞膜、IF16与DSPE-PEG-miR-181a的质量比为1：1：1。
6.根据权利要求1所述的制备方法，其特征在于，步骤(2)，所述超声的功率为200W、频率为28KHz，超声的温度为25℃，超声的时间为1h。
7.根据权利要求1所述的制备方法，其特征在于，步骤(2)，所述过滤为过滤膜为200nm的脂质体挤出机。
8.根据权利要求1所述的制备方法，其特征在于，步骤(2)，所述透析为使用孔径50nm的聚碳酸酯膜进行透析。
9.由权利要求1～8任一项所述的制备方法制备得到的miR181A@EM-Hb-IFI6材料。
10.权利要求9所述的miR181A@EM-Hb-IFI6材料在促进RISI愈合中的应用。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015052527A1 (en) * | 2013-10-09 | 2015-04-16 | Reneuron Limited | Microparticles, mirna and wound therapy |
| CN109414459A (zh) * | 2016-03-24 | 2019-03-01 | 斯坦姆实验室 | 源自脐带血的外泌体用于组织修复的用途 |
| CN111000980A (zh) * | 2019-12-06 | 2020-04-14 | 四川大学华西第二医院 | 干扰素诱导蛋白6的应用及药物 |
| CN113041224A (zh) * | 2019-12-26 | 2021-06-29 | 复旦大学 | 一种纳米人工红细胞及其在制备治疗细菌感染药物中的用途 |
| CN114948861A (zh) * | 2022-05-26 | 2022-08-30 | 中国人民解放军西部战区总医院 | 一种促进放射性皮肤损伤愈合的多功能水凝胶及其制备方法和应用 |
-
2022
- 2022-06-17 CN CN202210687844.2A patent/CN115177744A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015052527A1 (en) * | 2013-10-09 | 2015-04-16 | Reneuron Limited | Microparticles, mirna and wound therapy |
| CN109414459A (zh) * | 2016-03-24 | 2019-03-01 | 斯坦姆实验室 | 源自脐带血的外泌体用于组织修复的用途 |
| CN111000980A (zh) * | 2019-12-06 | 2020-04-14 | 四川大学华西第二医院 | 干扰素诱导蛋白6的应用及药物 |
| CN113041224A (zh) * | 2019-12-26 | 2021-06-29 | 复旦大学 | 一种纳米人工红细胞及其在制备治疗细菌感染药物中的用途 |
| CN114948861A (zh) * | 2022-05-26 | 2022-08-30 | 中国人民解放军西部战区总医院 | 一种促进放射性皮肤损伤愈合的多功能水凝胶及其制备方法和应用 |
Non-Patent Citations (2)
| Title |
|---|
| DAIJUN ZHOU, 等: "An injectable miR181a-IFI6 nanoparticles promote high-quality healing of radiation-induced skin injury", MATERIALS TODAY ADVANCES, vol. 15, 30 June 2022 (2022-06-30), pages 1 - 12 * |
| 张笑天;柴益民;: "微小RNA介导慢性创面愈合机制研究进展", 国际骨科学杂志, vol. 37, no. 06, 25 November 2016 (2016-11-25), pages 393 - 400 * |
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