咪喹莫特诱导小鼠银屑病样皮损机制的研究进展

咪喹莫特作为治疗人乳头瘤病毒感染的药物,在临床应用中发现可引起银屑病样皮损并能加重银屑病患者病情,而被广泛用于银屑病发病机制及治疗靶点的研究.由咪喹莫特诱发的小鼠银屑病样模型具有操作简单、易于成模的优点.应用咪喹莫特后,小鼠体内有包括Toll样受体信号通路等多条通路被激活,其中部分为协同作用.激活的信号通路进一步诱导角质形成细胞、树突细胞、Th17、γδT细胞、中性粒细胞等产生一系列炎性因子(如白细胞介素1、23、17,干扰素α等),这些炎性因子对银屑病样皮损的形成起到重要的作用.     

Notch信号通路相关配体在咪喹莫特诱导的银屑病样小鼠模型中的表达

目的：检测Notch信号通路相关配体在咪喹莫特（IMQ）诱导的银屑病样小鼠模型中的表达。方法： 6-8周雌性BALB/c小鼠随机分为IMQ模型组及空白对照组。HE染色观察皮损病理变化。RT-PCR及免疫组化分别检测IMQ模型组及空白对照组皮肤受体Notch1、Notch2、配体Jagged-1、目标基因Hes-1的mRNA和蛋白表达水平。结果：IMQ模型组小鼠皮肤出现典型银屑病样改变。IMQ模型组小鼠皮损中受体Notch 1、Notch2、配体Jagged-1、目标基因Hes-1的mRNA和蛋白表达水平高于对照组（P<0.05）。结论：Notch信号通路在咪喹莫特诱导的银屑病样小鼠模型中被激活。     

咪喹莫特诱导的银屑病样小鼠模型的组织学及免疫学特征初步探究

目的:研究咪喹莫特诱导的银屑病样小鼠模型的皮肤组织学及免疫学特征。方法:将20只BALB/c小鼠随机分为造模组和对照组,每组10只。所有小鼠背部剃毛后,造模组小鼠于背部皮肤及单侧耳部每日涂抹共计62.5 mg的5%咪喹莫特乳膏1次,连续7 d,空白组小鼠涂抹等量凡士林。每日记录小鼠PASI评分及耳部厚度,并于末次涂药24 h后处死,取背部皮肤进行多聚甲醛固定,行HE染色及Ki-67免疫组化分析。取脾脏称重,以流式细胞术检测脾脏中的Th细胞亚群。结果:造模组小鼠于第二天逐渐出现红斑、鳞屑、皮肤增厚及耳部肿胀,其PASI评分随造模时间延长而增加。造模组小鼠皮损出现银屑病样病理改变,其表皮厚度明显增加(t=15.12,P<0.001),且Ki-67阳性率明显上升,每个高倍镜视野下阳性细胞数目明显增加(t=10.50,P<0.001)。造模组小鼠脾脏重量和细胞计数较对照组明显增加,且Th1、Th17细胞亚群的比例及绝对计数较对照组明显上升(P值均<0.05),Th2细胞比例及计数无明显变化(P值均>0.05)。结论:咪喹莫特诱导的银屑病样小鼠模型可较好地模拟寻常型银屑病的临床、组织病理和免疫学特征,是一种有效的银屑病模型。     

芹菜素对咪喹莫特诱导的小鼠银屑病样皮损的影响及机制

目的:观察芹菜素对咪喹莫特(IMQ)造模小鼠及对角质形成细胞(HaCaT细胞)炎症通路和细胞增殖的影响.方法:小鼠随机分为4组,连续6 d使用凡士林或IMQ涂抹背部皮肤建立对照组和银屑病模型,期间每日予芹菜素或玉米油灌胃.于第7天比较治疗后各组皮损变化;蛋白免疫印迹法(western blot)检测各组核因子κB抑制蛋白(IKB)α、磷酸化(phosphoryla-tion,p)-IKBα、凋亡及细胞核因子-κB p65(NF-KB p65)、p-NF-KB p65、信号转导与转录激活因子(STAT)3、p-STAT3、细胞增殖抗原(Ki-67)和角蛋白(keratin,K)17蛋白变化.体外实验设对照组、模型组、5μmol/L芹菜素处理组和10 μmol/L芹菜素处理组;western blot 检测各组角质形成细胞 p-STAT3、STAT3、p-IKBα、IKBα、p-NF-KBp65、NF-KB p65、Ki-67 和 K17 变化.结果:芹菜素改善了 IMQ诱导的银屑病样小鼠的皮损及病理改变.与造模组比较,干预组小鼠皮损中p-STAT3、p-IκB、p-NF-κB p65、Ki-67、K17和NOD样受体热蛋白结构域相关蛋白(NLRP)3表达量显著下降(P＜0.05).细胞模型中,与模型组相比,5μmol/L 芹菜素显著抑制(P＜0.05)p-IKB α、p-NF-κBp65、Ki-67和K17的表达;10μmol/L芹菜素显著抑制(P＜0.05)p-STAT3、p-IKB α、p-NF-KB p65、Ki-67和K17的表达.芹菜素以剂量依赖方式抑制NLRP3的表达.结论:芹菜素可以缓解IMQ诱导的小鼠银屑病样皮损.在小鼠和细胞模型中,芹菜素对STAT3的磷酸化、NF-kappa B通路的激活、Ki-67的表达和K17的表达有明显的抑制作用,其可能通过抑制NLRP3的表达实现.     

人脐带间充质干细胞对咪喹莫特诱导的银屑病样皮炎小鼠模型的治疗作用

目的 研究人脐带间充质干细胞(MSC)对咪喹莫特诱导的银屑病样皮炎小鼠模型的治疗作用及机制.方法 将18只C57BL/6小鼠按随机数字表等分为凡士林组、模型组和治疗组,每组6只.小鼠背部剃毛后,模型组和治疗组小鼠背部涂抹5％咪喹莫特乳膏62.5 mg每天1次,连续6d,凡士林组小鼠涂抹等量的凡士林软膏;治疗组小鼠在第1...     

外用紫草素对咪喹莫特诱导银屑病样小鼠模型的干预作用及对CEBPD表达的影响

目的:探讨外用紫草素对咪喹莫特诱导的银屑病样小鼠模型的干预作用及对CCAAT增强子结合蛋白δ（CEBPD）表达的影响。方法:将20只SPF级BALB/c雄性小鼠按照简单随机抽样法分为模型组、紫草素1组、紫草素2组及空白对照组,每组5只。模型组、紫草素1组和紫草素2组小鼠背部脱毛区均每日外涂5%咪喹莫特乳膏（IMQ）50...     

抑制树突细胞MMP-13基因对咪喹莫特诱导的小鼠银屑病样炎症模型的影响

目的：明确抑制树突细胞MMP-13基因对咪喹莫特诱导的小鼠银屑病样炎症模型的影响。方法：构建树突细胞MMP-13基因特异性敲除的小鼠模型（MMP-13^LOX/LOX）,应用咪喹莫特诱导小鼠银屑病样皮损的发生。将小鼠分为野生型对照组、MMP-13^LOX/LOX对照组、咪喹莫特诱导的野生型实验组和咪喹莫特诱导的MMP-13^LOX/LOX实验组,分析不同组别小鼠皮损临床表现并记录PASI评分,检测各组小鼠皮损中IL-1β、IL-6、IL-23和IL-17A表达。结果：与咪喹莫特诱导的野生型实验组小鼠比较,咪喹莫特诱导的MMP-13^LOX/LOX实验组银屑病皮损表现相比更加温和,仅有轻度的棘层肥厚和真皮细胞浸润,皮损中IL-1β、IL-6表达下调,而IL-23和IL-17A表达无明显改变。结论：抑制树突细胞MMP-13基因可降低炎症因子IL-6、IL-1β的表达及减弱小鼠皮肤银屑病炎症反应。     

清热凉血方对咪喹莫特诱导的银屑病样小鼠模型JAKs/STATs通路表达影响的研究

目的 观察清热凉血方对咪喹莫特诱导的银屑病样小鼠模型皮损中酪氨酸激酶/信号转导因子和转录激活因子(JAKs/STATs)信号通路表达的影响.方法 BALB/c雄性小鼠48只,随机分为空白组、模型组、雷公藤多苷组及清热凉血方低、中、高浓度组,每组8只,采用外用咪喹莫特乳膏诱导产生银屑病样皮损.肉眼观察小鼠背部皮损严重程度...     

补体C3调控细胞凋亡抑制咪喹莫特诱导小鼠银屑病样皮肤炎症反应

目的 利用咪喹莫特(IMQ)诱导小鼠银屑病样皮炎模型,探讨补体C3在该病理过程中的作用及机制。方法 建立IMQ诱导银屑病样皮炎模型,比较C3^+/+及C3^-/-小鼠皮炎程度及病理改变;IHC检测皮损表皮细胞增殖,T细胞、中性粒细胞及巨噬细胞浸润,凋亡相关蛋白表达;TUNEL检测皮肤细胞凋亡;FCM比较小鼠皮损局部白细胞及中性粒细胞浸润及腋窝引流淋巴结IFN-γ应答。Z-VAD-FMK阻断凋亡途径,检测上述指标变化。结果 C3^-/-小鼠皮炎较C3^+/+组明显加重,表现为：角质形成细胞增生、微脓肿、棘皮症及炎症细胞浸润等显著增加(HE),角质形成细胞异常增殖(Ki67)、T细胞(CD3和CD4)、中性粒细胞(Ly-6G)及巨噬细胞(F4/80)浸润明显增加(IHC),凋亡相关蛋白(Cleaved casp3、Cyt C及BAK)表达显著上调(IHC),TUNEL阳性凋亡细胞明显增加(IF)。此外,FCM发现C3^-/-小鼠皮损局部白细胞(CD45)及中性粒细胞(Gr-1)浸润较C3     

DnaJA4敲除加重咪喹莫特诱导的小鼠银屑病样皮肤炎症

目的 探讨DnaJA4对咪喹莫特诱导小鼠银屑病样皮肤炎症的影响.方法 4组小鼠(每组8只),即应用咪喹莫特(imiquimod,IMQ)诱导银屑病样皮炎的DnaJA4敲除(knockout,KO-IMQ)组小鼠及野生型(wild-type,WT-IMQ)小鼠,涂以等量凡士林的DnaJA4敲除(knockout,KO-b...     

Mild electrical stimulation with heat shock reduces inflammatory symptoms in the imiquimod‐induced psoriasis mouse model

Psoriasis is a chronic skin disease caused by immune disorder. The chronic skin inflammation involves inflammatory molecules that are released from T lymphocytes and keratinocytes. Therefore, developing an anti‐inflammatory therapy that is suitable for long‐term treatment is needed. Electrical stimulation induces biological responses by modulating intracellular signaling pathways. Our previous studies showed that the optimized combination treatment of mild electrical stimulation (MES, 0.1‐millisecond; ms, 55‐pulses per second; pps) and heat shock (HS, 42°C) modulates inflammatory symptoms of metabolic disorders and chronic kidney disease in mice models and clinical trials. Here, we investigated the effect of MES+HS treatment on imiquimod‐induced psoriasis mouse model. Topical application of imiquimod cream (15 mg) to mice ear induced keratinocyte hyperproliferation and psoriasis‐like inflammation. In MES+HS‐treated mice, imiquimod‐induced skin hyperplasia was significantly decreased. MES+HS treatment reduced the protein expression of IL‐17A and the infiltration of CD3‐positive cells in lesioned skin. In addition, MES+HS‐treated mice had decreased mRNA expression level of antimicrobial molecules (S100A8 and Reg3γ) which aggravate psoriasis. In IL‐17A‐stimulated HaCaT cells, MES+HS treatment significantly lowered the mRNA expression of aggravation markers (S100A8, S100A9 and β‐defensin2). Taken together, our study suggested that MES+HS treatment improves the pathology of psoriasis via decreasing the expression of inflammatory molecules.     

银屑病动物模型研究进展

构建理想的银屑病动物模型是银屑病发病机制和开展药物治疗研究的关键,本文就银屑病模型的诱导,包括普萘洛尔诱导、咪喹莫特诱导、十二烷基硫酸钠诱导及细胞因子注射诱导和模型特点及局限性进行综述。     

Poly(ADP-ribose) polymerase-1 depletion enhances the severity of inflammation in an imiquimod-induced model of psoriasis

Poly(ADP-ribose) polymerase-1 (PARP1) is a pro-inflammatory protein, whose pro-inflammatory properties were demonstrated in human. The pro-inflammatory properties of PARP1 were shown in Th1- and Th2-mediated inflammatory pathologies, but not Th17-mediated inflammation. Thus, we studied the role of PARP1 in the imiquimod-induced model of psoriasis. To our surprise, in imiquimod-induced psoriasis, PARP1 acted as an anti-inflammatory factor and its genetic deletion exacerbated symptoms. We showed that in the absence of PARP1, the epidermis thickened and the number of TUNEL-positive cells decreased in the epidermis. These data indicate programmed cell death is decreased in keratinocytes. Changes in involucrin expression suggest that keratinocyte differentiation is hampered. Furthermore, epidermal expression of IL6 increased in the psoriasiform lesions of PARP1 knockout mice, suggesting that the inflammatory response is also derailed in the absence of PARP1. Finally, we showed that PARP1 expression is reduced in human psoriatic lesions compared with control skin samples. In imiquimod-treated HPV-KER keratinocytes, PARP inhibition recapitulated the in vivo findings, namely keratinocyte hyperproliferation; furthermore, the mRNA expression of psoriasis-associated cytokines (IL6, IL1β, IL8, IL17 and IL23A) was also induced. The inhibition of TRPV1 abrogated the effects of the combined imiquimod + PARP inhibitor treatment.     

Osteopontin deficiency affects imiquimod‐induced psoriasis‐like murine skin inflammation and lymphocyte distribution in skin,draining lymph nodes and spleen

Osteopontin (OPN) that enhances autoimmunity is expressed in psoriasis lesions; however, its functions in psoriatic inflammation are unknown. We investigated the role of OPN in OPN deficient mice (OPN?/?) by inducing psoriasis‐like inflammation through skin application of imiquimod (IMQ). OPN?/? mice treated with IMQ showed delayed onset ear swelling and attracted less inflammatory cells to the skin. IMQ‐induced lymph node swelling was reduced in the absence of OPN, and IMQ‐mediated expansion of B cells was inhibited. Further, reduction of CD4⁺ T‐cell numbers by IMQ in lymph nodes was suppressed in OPN?/? mice, with an increase in the CD4/CD8 ratio. A comparable pattern was found in spleen. Importantly, IMQ‐induced IL‐17 and IL‐4 expression by CD4⁺ lymph node T cells was reduced in OPN?/? mice. In conclusion, OPN may modulate psoriasis‐like inflammation through altering lymphocyte distribution in skin and draining lymph nodes and by inducing IL‐17 expression of inflammatory T cells.     

Mustard seed (Sinapis Alba Linn) attenuates imiquimod‐induced psoriasiform inflammation of BALB/c mice

Psoriasis is a chronic inflammatory autoimmune disease with undefined etiology. All present treatments are symptomatic. The unsatisfactory outcome in the treatment of psoriasis is partially due to the poor compliance to the present therapies with more or less side‐effects. As is known, drug homologous food is a popular intervention of some chronic diseases in Chinese traditional medicine. Mustard seed, consumed largely as a spice and a medicine in China, has recently been found to possess the bioactivities of anti‐oxidation, anti‐inflammation and anticancer. Therefore, it was supposed that mustard seed may have effects on psoriasis, and it was preliminarily validated using a BALB/c mouse model of psoriasiform inflammation induced by the topical application of imiquimod cream (Aldara) for 6 days consecutively. It was found that the forage containing 5% mustard seed obviously attenuated imiquimod‐induced psoriasiform inflammation, but did not clear it completely, accompanied by reduced infiltrations of T cells, plasmacytoid dendritic cells (pDC) and macrophages in lesional skin; reduced percentages of pDC and macrophages in the composition of immunocytes of spleens; reduced content of lesion nuclear factor‐κB p65, plasma malondialdehyde, lesion inducible nitric oxide synthase, interferon‐α, interleukin (IL)‐17 and IL‐22 at mRNA and protein levels; increased activities of superoxide dismutase, catalase and glutathione peroxidase; and increased percentage of CD4⁺ T cells and increased ratio of CD4⁺/CD8⁺ T cells in the composition of immunocytes of spleen. These results presented herein provide a basis for mustard seed to be used as a promising intervention for psoriasis in the future.     

Severe combined immunodeficiency mouse-human skin chimeras: a unique animal model for the study of psoriasis and cutaneous inflammation

Elucidation of the molecular and cellular mechanisms responsible for the pathogenesis of psoriasis had been significantly handicapped due to lack of an ideal animal model. To overcome this hurdle several investigators have developed a number of animal models for psoriasis. Recent establishment of the SCID-human skin chimeras with transplanted psoriasis plaques has opened new vistas to study the molecular complexities involved in psoriasis. This model also offers a unique opportunity to investigate various key biological events such as cell proliferation, angiogenesis, homing in of T cells in target tissues, neurogenic inflammation and cytokine/chemokine cascades involved in an inflammatory reaction. The SCID mouse model will be of immense help to target the cellular and molecular events associated with these pathogenic processes and develop novel drugs for psoriasis and other inflammatory diseases. In this article we have reviewed the prospects and the limitations of the SCID mouse model of psoriasis.     

Subclinical systemic and vascular inflammation detected by 18F‐fluorodeoxyglucose positron emission tomography/computed tomography in patients with mild psoriasis

Severe psoriasis is a systemic inflammatory disease involving major arteries and internal organs. The association between psoriasis and arterial inflammation, however, has been noted mostly in patients with moderate to severe psoriasis. Therefore, it is still not clear whether mild psoriasis also increases the arterial inflammation and cardiovascular disease. In this study, we aimed to investigate systemic inflammation using ¹⁸F‐fluorodeoxyglucose positron emission tomography/computed tomography (FDG‐PET/CT) in patients with mild psoriasis. In a nested case–control study, FDG‐PET/CT findings of 10 patients with mild chronic plaque psoriasis involving a body surface area of less than 5% were compared with those of 10 age‐ and sex‐matched healthy controls. The degree of FDG uptake in the large arteries (ascending aorta, aortic arch, descending aorta, suprarenal abdominal aorta and infrarenal abdominal aorta,) and liver were analyzed using the maximum target‐to‐background ratios (TBR). There were no significant demographic differences between the psoriasis patients and the control subjects (P > 0.05). Patients with psoriasis showed higher maximum TBR than healthy controls in all examined arterial segments, with statistical significance reached in the suprarenal abdominal aorta (P < 0.05), ascending thoracic aorta (P < 0.01) and infrarenal abdominal aorta (P < 0.05), and in the liver (P < 0.05). The “candy cane sign”, which represents typical arterial inflammation of the thoracic aorta, was noted upon PET in the majority of psoriasis cases. In conclusion, mild psoriasis is also associated with arterial and hepatic inflammation, which can be readily demonstrated by using FDG‐PET/CT.     

HLA-Cw6基因对生物制剂治疗银屑病疗效影响的Meta分析

目的：Meta分析评价HLA-Cw6基因阳性对生物制剂治疗银屑病疗效的影响。方法：检索1976年1月至2018年12月Pubmed、Embase、The Cochrane Library、中国生物医学文献数据库、万方数据库及中国知网等数据库中关于HLA-Cw6基因与生物制剂疗效关联的文献，对最终符合入选标准的文献进行Meta分析。结果：符合入选标准的文献共11篇，涉及三种生物制剂，包括：2项关于IL17A拮抗剂Secukinumab研究，3项抗肿瘤坏死因子α(TNF-α)拮抗剂的研究及6项IL-12/IL-23拮抗剂Ustekinumab的研究。对Ustekinumab疗效关联的6项研究Meta分析显示治疗后HLA-Cw6阳性患者中PASI评分下降75%的比例(PASI75)高于HLA-Cw6阴性患者，差异有统计学意义(P<0.05)。Secukinumab的2项研究及TNF-α拮抗剂治疗的3项研究Meta分析显示HLA-Cw6阳性患者中PASI75较HLA-Cw6阴性者无明显增高，差异无统计学意义(P>0.05)。结论：携带HLA-Cw6基因患者对IL-12/IL-23拮抗剂Ustekinumab的疗效优于未携带者，HLA-Cw6对Secukinumab和TNF-α拮抗剂疗效无明显影响。