Classification and research prospect of in vitro and in vivo models of psoriasis

doi:10.3969/j.issn.2095-4344.1395

Abstract

Abstract:

BACKGROUND: In vivo and in vitro models of psoriasis are an important way to understand the pathogenesis. Although existing animal models can partially simulate the pathological manifestations of psoriasis, they are not representative.
OBJECTIVE: To summarize the classification and research progress of animal and cell models of psoriasis, and to provide beneficial references for researchers to select appropriate models of psoriasis.
METHODS: PubMed, CNKI and WanFang databases were retrieved with the keywords of “psoriasis and animal model” in English and Chinese, respectively. After screening the literature, 51 eligible articles were finally included.
RESULTS AND CONCLUSION: According to the animal modeling method, the literature included in the study was summarized. The animal models used for psoriasis included genetic engineering mouse model, spontaneous psoriasis mouse model, exogenous induced psoriasis model, xenotransplantation model and traditional Chinese medicine disease-syndrome combination animal model. All kinds of animal models of psoriasis simulate the pathological manifestations of psoriasis to a certain extent, but there are still limitations of differences in pathogenesis and species. Cell models can reveal changes in biological behaviors and interactions of keratinocytes or other cell types in psoriasis. Researchers can choose the most suitable model of psoriasis in vivo and in vitro according to their research purposes and experimental condition.

Key words: psoriasis, animal model, cell model, classification, interleukin, genetic engineering

CLC Number:

Xu Yongyue1, Chen Haiming1, Deng Jingwen1, Wang Maojie1, 2, Yan Yuhong1, Huang Runyue1, 3, Lu Chuanjian1, 3. Classification and research prospect of in vitro and in vivo models of psoriasis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(27): 4414-4419.

Figures/Tables 2

2.1 动物模型由于自然界动物几乎不会自发出现类似银屑病皮损的临床表现，因此需要模拟其发病特点。目前，研究者或使用基因工程、或筛选基因突变鼠、或给予外源刺激等手段去构建银屑病小鼠，以期一定程度上符合其病理特征。 2.1.1 基因工程小鼠模型基因工程小鼠是研究特定基因对疾病发病机制的重要模式生物，利用基因工程技术(如条件基因打靶、基因捕获、RNA干扰等)，可实现目的基因的转入、敲除/沉默。以银屑病转基因鼠为例，通常选择在K5或K14启动子插入目的基因全片段cDNA，然后将其显微注射进小鼠胚胎原核中，待后代繁殖发育后，行聚合酶链式反应(PCR)检测去筛选、鉴定基因型。 (1)白细胞介素1α转基因小鼠模型：通过对模型鼠(TgIL1.1)的研究证实，高表达白细胞介素1α基因使小鼠皮肤出现银屑病样改变，其中红斑、鳞屑多集中在胸部，病理特征包括非皮损处真皮单核/巨噬细胞浸润，皮损处混合淋巴细胞浸润。但伴随消瘦体质，11个月大的小鼠逐渐秃顶、体质量损失、皮肤变皱[5]。另一种IL1rn-/-小鼠模型由于白细胞介素1α受体拮抗剂缺乏，有约60%的小鼠耳郭皮肤出现类银屑病炎症[6]。除了皮肤炎症，该模型还会出现动脉炎症和关节炎，肿瘤坏死因子、白细胞介素36α在白细胞介素1α转基因鼠模型中发挥关键作用[7-8]。 (2)PL/J/CD18缺陷小鼠模型：该模型CD18(即β2整合素)表达降低，其在多种炎症反应中起重要作用。PL/J/CD18缺陷小鼠通常在出生11周后，会出现类银屑病皮损。有趣的是，在PL/JxC57BL/6J基因背景上的纯合突变F1代小鼠没有发展出银屑病，而回交实验(即F1代与任何亲本的交配)表明除了CD18之外，还有少量基因决定了疾病的易感性[9]。β2整合素主要与角质形成细胞过度增殖和分化异常有关。另有研究报道使用JAK抑制剂通过影响T细胞功能而显著减轻该模型皮损[10]。 (3)K5-IL-17C转基因小鼠：白细胞介素17C是白细胞介素17家族的成员之一，研究显示白细胞介素17C或白细胞介素17A均可诱导皮肤炎症反应，而白细胞介素17C是银屑病皮损部位表达最高的白细胞介素17蛋白。白细胞介素17C小鼠出生后8周后出现背部皮肤增厚、红斑和表皮脱落，病理特征显示CD4和CD8 T细胞、巨噬细胞和髓样树突状细胞局部浸润，血管生成增多。皮损处表达多种促炎症因子，而应用肿瘤坏死因子抑制剂后皮损明显消退，机制与NF-κB通路调节或白细胞介素17A的协同作用有关[11]。另一种K14-IL-17Aind/+小鼠由白细胞介素17Aind和K14-Cre等位基因杂交后产生，该模型会出现严重的银屑病样皮损[12]。研究其发病机制与活性氧生成增加和血液中循环炎性白细胞、内皮功能障碍、收缩增加有关[13]。 (4)K5.TGFbeta1wt转基因小鼠：转换生长因子β参与角质细胞增殖、血管生成、T细胞分化，在银屑病的发生发展过程中起作用。从第17天开始，转基因角质细胞在皮肤扩增，K5.TGFbeta1wt转基因小鼠逐渐出现严重的银屑病样皮损，血管生成伴血管扩张，多种趋化因子上调；该模型至6个月大时已全身皮肤红斑鳞屑，和人类的红皮型银屑病表现相似[14-15]。 (5)K5.Stat3C转基因小鼠：证实了信号转导因子和转录激活因子3(signal transducer and activator of transcription，STAT3)在角质形成细胞中的作用。转录激活因子3C缺陷小鼠会导致伤口愈合延迟，而K5.Stat3C小鼠则会出现红斑鳞屑，伴随伤口处的皮损肥厚。皮损处毛细血管数量增加，血管内皮生长因子、细胞间黏附分子1、IκB-α蛋白等表达上调，转录激活因子3C与活化的T细胞(特别是Th17细胞)一起，促进皮肤的炎症反应[16-18]。见表1。 2.1.2 自发银屑病样小鼠模型 (1)Scd1ab/Scd1ab纯合子小鼠：是首个出现角化过度的体内模型，其编码硬脂酰辅酶A去饱和酶的Scd1基因突变引起皮脂腺缺失，成年小鼠出现鳞屑、血管增加及巨噬细胞浸润等部分类似人类银屑病，缺点是该模型真皮也过度增长、成纤维密度增高；在皮损处缺乏T细胞和中性粒细胞的浸润，表皮增生可能由肥大细胞分泌的炎性递质引起，因而实际应用不广泛[24]。 (2)Ttcfsn/ Ttcfsn鼠：是常染色体隐性突变模型，该模型出生2周后会出现皮肤鳞屑、炎症、角化过度和Koebner阳性反应，并且皮损处中性粒细胞浸润。然而，该模型血中表皮生长因子受体和IgE抗体水平升高，且对环孢素治疗并无效果，加之相对寿命较短，因而其应用也受到限制[25]。 (3)Sharpincpdm/Sharpincpdm鼠：Hogen等[26]发现C57BL/Ka小鼠上的cpd突变，该模型会活化肿瘤坏死因子α诱导的NF-κB活性，出现多器官炎症包括腹背部慢性增生性皮炎的表现。该皮炎用皮质类固醇治疗有效，对环孢素A无效。但该鼠缺点是口腔、胃部也有类似上皮细胞增殖的病变，缺乏临床表型特异性[27]。除此之外，还有研究者利用小鼠尾部鳞片中表皮颗粒层作为模型，观察药物给药对表皮颗粒层的治疗作用[28]。其余几种基因突变品种小鼠(ic/ic突变鼠、hr/hr突变鼠等)，因其与银屑病病理相差较大，因而使用范围较少。 2.1.3 诱发银屑病样模型 (1)咪喹莫特(5%)诱导小鼠模型：咪喹莫特为Toll样受体(Toll-like receptor，TLR)7/8激动剂，激活固有免疫系统导致中性粒细胞、γδT细胞在皮损浸润，产生炎症因子、趋化因子等，促进或加重银屑病样皮损改变，同时伴随食欲减退、体质量损失、脾脏增大等全身表现。该模型小鼠通常诱导7 d后背部皮损改变最为严重，机制主要通过白细胞介素23/白细胞介素17通路介导[29-30]。该模型优点为操作简单、易于复制，成为国内研究的主要诱导银屑病小鼠模型；缺点为该模型具有自愈倾向，随着诱导时间延长，其背部皮损逐渐稳定或减轻，因此不太适于发病机制的长期研究。 (2)十二烷基硫酸钠(10%)诱导小鼠模型：背部皮肤涂抹十二烷基硫酸钠诱导7 d之后，小鼠出现皮肤增厚、局部红肿和皮下血管增生，病理改变包括角化过度、角质层增厚，真皮层和皮下毛细血管扩张和充血。通过检测血清蛋白显示蛋白激酶Cβ、白细胞介素1β、肿瘤坏死因子α、白细胞介素23等细胞因子表达上调。该小鼠模型反映了银屑病的部分特征，但国外应用此模型研究较少，其刺激的信号通路亦不甚清楚[31-32]。 (3)普萘洛尔诱导豚鼠模型：20世纪90年代，科学 "

家最初给豚鼠单纯灌胃普萘洛尔0.1 mg/d并没有诱导出银屑病样改变，但每周皮下注射完全弗氏佐剂之后能诱导成功[33]。后来经过逐渐改进造模方法，现多采用在豚鼠耳部涂抹5%普萘洛尔乳剂，几天后可观察到包括皮损局部红肿、糜烂，角化不全，毛细血管扩张等病理改变，皮损部位多种促炎因子(如白细胞介素)表达增加[34-35]。 (4)皮内注射白细胞介素23诱导模型通过向小鼠皮内注射1 μg量的白细胞介素23刺激产生白细胞介素19和白细胞介素24，它们通过肿瘤坏死因子信号途径影响角质形成细胞的分化和增殖。小鼠会出现皮损红斑、角化不全、表皮增生等表现，伴随CD4+淋巴细胞、树突状细胞、中性粒细胞和巨噬细胞浸润，与许多银屑病的显著症状相吻合[36]。此外，尚有研究体内注射雌激素刺激小鼠阴道上皮组织增生来模拟银屑病皮损表现，该小鼠上皮细胞的增殖核抗原表达水平对照组升高，阴道上皮基底细胞的有丝分裂指数增高[37]。 2.1.4 异种移植银屑病模型异种移植模型是目前最接近人类银屑病改变的模型，它对银屑病发病机制和新药研发方面有重要价值。异种移植模型主要使用严重免疫缺陷小鼠作为受体，亦有研究使用裸鼠或AGR129鼠为受体[38]。国外学者将银屑病受累皮肤移植至严重免疫缺陷鼠，10周之后再将自体皮损处T细胞(而非外周血T细胞)注射进移植的真皮内，可观察到银屑病病理改变的维持[39]。该模型优点是代表性较好，缺点是需要大量银屑病患者供体皮肤，供体来源受到限制，同时还要考虑供体间的遗传异质性。此外，该模型价格昂贵且对饲养方面要求较高，限制了其推广应用。 2.1.5 中医病证结合动物模型(银屑病) 目前，经过研究报道的银屑病病证结合动物模型共有3种，涉及脾虚证、湿热证和血瘀证。银屑病中医病证结合动物模型均采用2种或多种因素复合造模，步骤较多，造模周期从1周到3周不等。经过造模后，研究报道这些方法均能够一定程度上模拟特定的银屑病病证结合的临床表现，如脾虚证豚鼠除了银屑病皮损，还会出现体质量增长变缓、纳呆少饮、嗜睡懒动、大便时干时溏，肛温升高、血脂水平升高，类似人的脾虚证表现[40-41]。但是，由于人的中医证候指标与豚鼠/小鼠的物种差异，国内学界对脾虚证、湿热证和血瘀证的指标选择尚无统一的认识，有待进一步的研究证实病证结合模型与临床指标之间的关联。具体病证结合模型分类见表2。 2.2 细胞模型 2.2.1 二维(2-Dimensional，2D)细胞模型角质形成细胞是银屑病研究最为常见的2D细胞类型。银屑病皮损角质形成细胞呈过度增殖和异常分化，其诱发过程可能与接受皮损局部的易感因素(基因易感性、炎症微环境等)有关。从银屑病皮损处分离、体外培养角质形成细胞虽然代表性好，但这种方法可重复性、可获得性和稳定性都较差，并且其银屑病表型的维持需要添加细胞因子。因此，目前最常用的细胞有正常表皮角质形成细胞和 "

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