Apigenin ameliorates imiquimod-induced psoriasis in C57BL/6J mice by inactivating STAT3 and NF-κB

Psoriasis is a chronic autoimmune disease featured by patches on the skin.It is caused by malfunction of immune cells and keratinocytes with inflammation as one of its key features.Apigenin(API)is a natural flavonoid with anti-inflammatory and immunoregulatory properties.Therefore,we speculated that API can ameliorate psoriasis,and determined its effect on the development of psoriasis by using imiquimod(IMQ)-induced psoriasis mouse model.Our results showed that API attenuated IMQ-induced phenotypic changes,such as erythema,scaling and epidermal thickening,and improved splenic hyperplasia.Abnormal differentiation of immune cells was restored in API-treated mice.Mechanistically,we revealed that API is a key regulator of signal transducer activator of transcription 3(STAT3).API regulated immune responses by reducing interleukin-23(IL-23)/STAT3/IL-17A axis.Moreover,it suppressed IMQ-caused cell hyperproliferation by inactivating STAT3 through regulation of extracellular signal-regulated kinase 1/2 and nuclear factor-κB(NF-κB)pathway.Furthermore,API reduced expression of inflammatory cytokines through inactivation of NF-κB.Taken together,our study demonstrates that API can ameliorate psoriasis and may be considered as a strategy for psoriasis treatment.

Coleus forskohlii shows anti-psoriatic activity in imiquimod-induced psoriasis rats

Objective:To evaluate anti-psoriatic activity of Coleus forskohlii in rats with imiquimod-induced psoriasis.Methods:Imiquimod was used to induce psoriasis in rats.Body weight,skin thickness,erythema,scaling,spleen weight,and histological alternations were measured to assess the effect of Coleus forskohlii.Furthermore,an emulgel formulation containing Coleus forskohlii 10%was prepared and characterized along with its ex vivo permeation studies.Results:The emulgel formulation containing Coleus forskohlii 10%had a pH of 5.40±0.36,with optimum spreadability of(31.67±2.08)g/(cm·s)and viscosity of(15966.67±1274.10)cps,and enhanced both the rate and the extent of drug permeation through psoriatic skin.In an ex vivo study,the quantity of drug permeated(19.18%),deposited(52.38%),and drug remaining in the donor compartments(28.31%)was satisfactory.Coleus forskohlii significantly alleviated imiquimod-induced psoriasis by increasing glutathione and superoxide dismutase activity,decreasing malondialdehyde and nitric oxide levels,and alleviating histological alternations in rat skin.Conclusions:Coleus forskohlii can alleviate imiquimod-induced psoriasis,which may be used as a therapeutic candidate for the treatment of psoriasis.

Imiquimod:A potential option for inhibition of repigmentation of congenital melanocytic nevus after laser ablation

Congenital melanocytic nevi(CMN) are common skin tumors. Large and specially located nevi cannot be completely removed by surgery, posing the risks of both cosmetic deformities and potential malignancy.Nonsurgical treatments, such as laser therapy and physical dermabrasion, can overcome the limitations of surgery;however, the high rate of repigmentation remains an unresolved global challenge. We conducted a self-controlled observational study of a patient with a nevus on the chest. Two areas of the lesion were treated with an Er:YAG laser and 5% imiquimod cream was applied to one of these areas. After nearly 7-months of follow-up, we observed a significant difference in color between the two areas, suggesting that topical imiquimod may inhibit repigmentation and significantly enhance the effectiveness of laser treatment.

树突状细胞疫苗联合Imiquimod对小鼠乳腺肿瘤的治疗作用

目的观察局部应用TLR7激动剂Imiquimod对树突状细胞(dendritic cell,DC)疫苗治疗小鼠肿瘤的影响,寻找增强DC肿瘤疫苗疗效的佐剂。方法培养乳腺肿瘤细胞株EMT-6,反复冻融法制备肿瘤相关抗原,小鼠乳腺区注射EMT-6制备荷瘤动物模型。用含重组小鼠GM-CSF和IL-4的培养液扩增培养Balb/c小鼠来源的BM-DC,负载肿瘤相关抗原(Ag-DC)后进行肿瘤治疗实验。荷瘤小鼠分别采用Imiquimod局部涂抹、Ag-DC皮内注射、Imiquimod联合Ag-DC进行治疗,检测不同处理方法小鼠的肿瘤生长情况。采用上述方案免疫Balb/c小鼠后细胞增殖抑制实验检测小鼠脾脏淋巴细胞对EMT-6细胞的杀伤效应。流式细胞术检测Imiquimod处理局部皮肤对BM-DC向局部引流淋巴结(DLN)迁移的影响。结果 Imiquimod联合Ag-DC治疗荷瘤小鼠可明显抑制肿瘤生长,与单纯Imiquimod、Ag-DC治疗组及NS处理对照组比较差异具有显著性。Imiquimod联合Ag-DC免疫小鼠后脾脏淋巴细胞杀伤肿瘤细胞能力较其它组明显增强。局部涂抹Imiquimod后注射BM-DC,24 h后DLN内BM-DC数量较对照组明显增多。结论 TLR7激动剂Imiquimod可增强BM-DC疫苗诱导的免疫应答,提高肿瘤治疗效果。其机制可能与Imiquimod诱导DLN BM-DC细胞数量增多有关。

咪喹莫特(imiquimod)联合树突状细胞降低荷黑素瘤小鼠调节性T细胞并增强抗肿瘤效果

目的观察联合应用Toll样受体7(TLR7)激动剂咪喹莫特(imiquimod)和负载肿瘤相关抗原的树突状细胞(DC)疫苗对荷黑素瘤小鼠的治疗作用并探讨相关机制。方法培养表达卵白蛋白的B16黑素瘤(B16-OVA)细胞,注射入C57BL/6小鼠皮下制备荷瘤小鼠模型。含重组小鼠粒细胞巨噬细胞集落刺激因子(rm GM-CSF)和重组小鼠白细胞介素4(rm IL-4)的培养液扩增培养骨髓来源的DC,加入OVA过夜孵育制备负载OVA的DC疫苗(OVA-DC)。荷瘤小鼠分别采用PBS、咪喹莫特局部涂抹、OVA-DC皮内注射、咪喹莫特联合OVA-DC进行治疗,数字游标卡尺测量小鼠皮下肿瘤的生长情况。流式细胞术检测荷瘤小鼠外周血CD4^+FOXP3^+调节性T细胞(Treg)的比例。采用上述方案免疫小鼠后,用CCK-8法检测小鼠脾脏淋巴细胞对B16-OVA细胞的杀伤效应。结果与单纯咪喹莫特、OVA-DC治疗组及PBS对照组相比,咪喹莫特联合OVA-DC治疗组荷瘤小鼠黑素瘤体积较小,荷瘤小鼠CD4^+细胞中FOXP3^+Treg比例明显降低。咪喹莫特联合OVA-DC免疫小鼠脾脏淋巴细胞杀伤B16-OVA肿瘤细胞能力较其他组明显增强。结论咪喹莫特联合负载抗原的DC疫苗可诱导荷瘤机体CD4^+FOXP3^+Treg比例降低并增强抗肿瘤效果。

Imiquimod和Resiquimod在皮肤科的应用

新型小分子免疫反应调节剂Imiquimod(R-837)和Resiquimod(R-838)结构相似,具有抗病毒和抗肿瘤活性,可用作B淋巴细胞活化剂和免疫佐剂,在皮肤科广泛应用于治疗病毒性皮肤病及各种良恶性肿瘤。

Imiquimod治疗9例外生殖器尖锐湿疣的临床报告

目的观察用i miqui mod治疗9例外生殖器尖锐湿疣的疗效及相关问题。方法5%i mqui mod霜睡前使用1次,每周3次,致疣体消失为止,疗程不超过16周。结果5例疣体在用药5周后完全消失,2例在6周后消失,2例在用药4周后改用电灼术去除疣体。患者均有局部药物不良反应,可耐受。2例治疗结束后2个月复发。结论I miqui mod可用于治疗外生殖器尖锐湿疣。

Clinical observations on the treatment of infantile hemangiomas with topical imiquimod 5% cream

Objective： To observe the efficacy and safety of topical imiquimod 5% cream in the treatment of uncomplicated infantile hemangiomas （IHs）. Methods： A total of 68 IHs were treated with topical imiquimod 5% cream. Among them, 36 were superficial, 22 were mixed, and 10 were deep. The size of IHs ranged from 1.0 cm × 1.5 cm to an area of a whole forearm. All the hemangiomas were in a proliferative stage. Imiquimod was applied 3 times weekly in 44 patients and 5 times weekly in 24 patients for up to 36 weeks. Results： All superficial IHs improved, and 18 achieved complete clinical resolution, 10 had excellent improvement, 5 showed moderate improvements, and 3 patients displayed minimal improvement. Two mixed IHs showed excellent improvement, 3 showed moderate improvement and 5 manifested minimal improvements. The remaining 12 mixed IHs and all deep IHs did not respond to the therapy. The total incidence of local adverse events was 58.82%（40/68）, which included erythema or edema, local itching, incrustation or peeling, erosion or ulceration, although most of these were mild to moderate reactions and did not affect the treatment. Scarring occurred in 2 mixed IHs. No systemic side effects developed. Conclusion： Imiquimod 5% cream may be a safe and effective alternative for the treatment of superficial IHs and some mixed IHs in which the superficial component predominates. An appropriate treatment duration for proliferative IHs treated with this therapy may be 24 weeks. Some local adverse events, such as crusting and erosion with possible scarfing potential may occur and should be addressed by prompt, but temporary, discontinuation of the imiquimod. Topical imiquimod 5% cream can be prudently used in the treatment of IHs larger than 5.0 cm × 5.0 cm in newborns and infants less than 6 months of age. To our knowledge, this is the largest IH group treated with imiquimod that has been reported in the literature to date.

免疫调节剂Imiquimod

免疫调节剂ＩｍｉｑｕｉｍｏｄＩｍｉｑｕｉｍｏｄ的化学名为１－异丁基－１Ｈ－咪唑［４，５－Ｃ］喹啉－４胺，曾用代号为Ｓ－２６３０８。它是免疫调节剂，也是干扰素诱导剂（Ｉｎｔｅｒ－ｆｅｒｏｎＩｎｄｕｃｅｒ）。最初由美国的ＲｉｋｅｒＬａｂｓ研制开发，并于８...

Holistic Approach in the Treatment of Actinic Keratosis: Benefits and Disadvantages of 5-Fluorouracil, Imiquimod, Diclofenac and Curaderm

Background Actinic keratosis is the most prevalent premalignant skin disorder in the white population. Current guidelines provide no clear recommendations about preferred treatments. Methods The parameters;effectiveness, treatment duration, recurrence, side effects and cost of treatment were investigated for three frequently used topical therapies which were then compared with a most recent developed topical therapy. Published clinical data obtained from the literature was used to compare these parameters for 5-fluorouracil, imiquimod and diclofenac and relate them with the newly developed Curaderm. Results A wide variation in the concentrations of the active anti-keratotic ingredients, application frequency, duration of treatment, recurrence rates and cost of treatment exist between the different topical therapies. The efficacy rates and side effects were less variable. Overall, Curaderm is the most suitable treatment for actinic keratosis. Clinical evidence is presented illustrating the effects of Curaderm on field-directed treatments and solitary treatments of actinic keratoses. Conclusions Current medical guidelines do not provide clear recommendations on which treatment approach for actinic keratosis is preferred. Direct head-to-head comparison between treatments with emphasis on efficacy, safety, treatment duration, compliance, convenience, cosmetic outcome, patient acceptance and cost should be available to the patient, the practising physician, healthcare system and should assist in therapeutic treatment guidelines and policymaking. Given the very favourable profiles of these parameters with Curaderm when compared with other home-based treatments, it should be considered that Curaderm is first-in-line.

Successful Treatment of Suborbital Pyogenic Granuloma with Topical Imiquimod

Pyogenic granuloma (PG) is a benign vascular proliferation of the skin and mucosae, that has been treated with different regimens with variable success and recurrence rates;however, the management of PG still remains a challenging issue, particularly in children and in adult cases with lesions localized at sites difficult to access. Imiquimod, a member of the imidazoquinoline family of immune response modifiers, is a topically applicable TLR-7/8 agonist that reveals potent antiviral, antitumor, immunoregulatory and antiangiogenic properties. In the present paper we report the case of a 9-year old boy with suborbital pyogenic granuloma, successfully treated with topical daily application of imiquimod 5% cream without occlusion. 8 weeks after onset of topical imiquimod treatment a complete resolution of the lesion without any scarring was observed. No systemic side effects were seen and the patient remained well throughout the course of therapy. He is presently completing a 15-month follow-up and has revealed no relapse. The findings of the present study suggest that topical imiquimod is a safe, effective and well-tolerated treatment for PG in children, even at difficult to treat areas like the suborbital region.

Imiquimod 以 TLR7非依赖途径诱导THP-1来源的巨噬细胞凋亡

目的：探讨TLR7在imiquimod引起的THP-1来源的巨噬细胞凋亡中的作用。方法选择TLR7表达强度不同的细胞系（ THP-1巨噬细胞、MDCK细胞和HUVEC细胞），通过MTT法分别检测经不同浓度imiquimod处理后细胞的存活率。酶联免疫吸附试验检测通过TLR7抑制剂氯喹、TLR7-siRNA处理减少TLR7表达后，细胞上清液中IL-6的水平。流式细胞术进一步检测抑制TLR7表达后，imiquimod对THP-1巨噬细胞凋亡的影响。结果 Imiquimod能引起THP-1巨噬细胞、MDCK细胞和HUVEC细胞这3种细胞的凋亡。氯喹和TLR7-siRNA处理减少TLR7表达后，能显著降低IL-6的水平，但不影响imiquimod引起的THP-1巨噬细胞凋亡。结论 Imiquimod可通过TLR7非依赖方式引起细胞凋亡。

Treatment of allergic airway inflammation and hyperrespon-siveness by imiquimod modulating transcription factors T-bet and GATA-3

Background Imiquimod is an imidazoquinoline, which class of compounds are known to have antiviral and antitumoural properties. In recent studies, it was shown that imiquimod modulates the T helper cell type Th1/Th2 response by inducing the production of Thl cytokines like IFN-γ, and by inhibiting the Th2 cytokines like interleukin （IL）-4. Several investigators have shown that T-bet and GATA-3 are master Thl and Th2 regulatory transcription factors. This study investigated whether irniquimod treatment inhibited airway inflammation by modulating transcription factors T-bet and GATA-3. Methods Thirty-six male SD rats were randomly divided into a control group, an asthmatic group, and an imiquimod group, which was exposed to an aerosol of 0.15% imiquimod. Twenty-four hours after the last ovalbumin （OVA） challenge, airway responsiveness was measured and changes in airway histology were observed. The concentrations of IL-4, IL-5 and IFN-γ in bronchoalveolar lavage fluid （BALF） and serum were measured by enzyme linked immunosorbent assay （ELISA）. The rnRNA expressions of IL-4, IL-5, IFN-γ, T-bet and GATA-3 in lung and in CD4^＋ T cells were determined by reverse transcription polymerase chain reaction （RT-PCR）. The protein expressions ofT-bet and GATA-3 were measured by Western blot. Results It was demonstrated that imiquimod 1） attenuated OVA induced airway inflammation; 2） diminished the degree of airway hyperresponsiveness （AHR）; 3） decreased the Th2 type cytokines and increased Thl type cytokines mRNA and protein levels; 4） modulated the Th1/Th2 reaction by inhibiting GATA-3 production and increasing T-bet production. Conclusion Imiquimod treatment inhibits OVA induced airway inflammation by modulating key master switches GATA-3 and T-bet that result in committing T helper cells to a Thl phenotype.

Imiquimod attenuates airway inflammation and decreases the expression of thymus and activation regulated chemokine in allergic asthmatic mice

Imiquimod is an imidazoquinoline derivative. Some studies have shown that imiquimod modulates the Th1/Th2 response by inducing the production of Th1 cytokines IFN-γ and interleukin （IL）-12. and by inhibiting Th2 cytokines IL-4 and IL-5. These data suggest that imiquimod has therapeutic appli . cations in atopic diseases such as allergic asthma that are associated with overexpression of Th2 cytokines.

Topical imiquimod treatment of cutaneous vascular disorders in pediatric patients:clinical evaluation on the efficacy and safety

Objective:To evaluate the clinical effect of topical imiquimod treatment on cutaneous vascular disorders in pediatric patients.Methods:A retrospective investigation was conducted in 25 pediatric patients with cutaneous vascular disorders,including 19 infantile hemangiomas(IHs)(12 superficial/7 mixed type),5 nevus flammeus(NF),and 1 pyogenic granuloma(PG).Imiquimod 5% cream was applied every other day for 4 to 16 weeks(average 9.6 weeks).Results:Of the 19 IHs treated,an overall efficacy of 52.6% was achieved,with a clinical resolution rate of 15.8%,excellent rate of 26.3%,and moderate rate of 10.5%.The superficial type responded the best at 66.7%,while the mixed type showed only 28.6% effectiveness,which was predominantly from their superficial parts.No obvious response was noted in the 5 patients with NF.Side effects were observed in 78.9% of the patients,mostly mild to moderate local irritations and occasionally severe reactions such as thick crusting and ulceration.Systemic side events were observed in 4 IH patients including fever and digestive tract reactions.No recurrence was observed during the follow-up examination.Conclusions:Topical imiquimod could be an alternative option for the treatment of uncomplicated superficial IHs with satisfactory tolerability.

Amelioration of imiquimod-induced psoriasis-like dermatitis in mice by DSW therapy inspired hydrogel

Psoriasis is a long-lasting and recurrent autoimmune disease which is incurable so far.Dead Sea water(DSW)therapy is an effective approach to help control the symptoms of psoriasis due to the abundant mineral ions in DSW,which inspired the material formulation in this study.Rubidium–Sodium alginate/Polyacrylamide hydrogels(Rb-SA/PAAm gels)composed of sodium alginate and polyacrylamide interpenetrating network structure with different concentrations of rubidium and certain magnesium and zinc content were prepared for the treatment of psoriasis.The obtained results suggest the good mechanical properties of the Rb-SA/PAAm gels including toughness and swelling performance.In terms of in vitro tests,the Rb-SA/PAAm gels not only show nontoxicity to human keratinocyte cell line(Hacats)but also inhibits the activity against inflammatory NF-κβ signaling pathway.Meanwhile,they can release Rb+which enable the Rb-SA/PAAm gels have better antibacterial ability to Streptococcus and Escherichia coli.The results obtained from in vivo tests indicate that these hydrogels could alleviate the symptoms of psoriasis caused by Imiquimod(IMQ)in mice by reducing the inflammatory factor in STAT3 pathway and therefore reduce the immune stimulation of the spleen.In conclusion,the 100Rb-SA/PAAm gel has demonstrated great potential to be a topical wettable dressing for psoriasis treatment.

Small interfering RNA targeting of keratin 17 reduces inflammation in imiquimod-induced psoriasis-like dermatitis

Background:Psoriasis is a common chronic inflammatory skin disease with 2%to 3%prevalence worldwide and a heavy social-psychological burden for patients and their families.As the exact pathogenesis of psoriasis is still unknown,the current treatment is far from satisfactory.Thus,there is an urgent need to find a more effective therapy for this disease.Keratin 17(K17),a type I intermediate filament,is overexpressed in the psoriatic epidermis and plays a critical pathogenic role by stimulating T cells in psoriasis.Therefore,we hypothesized that inhibiting K17 may be a potential therapeutic approach for psoriasis.This study aimed to investigate the therapeutic effect of K17-specific small interfering RNA(siRNA)on mice with imiquimod(IMQ)-induced psoriasis-like dermatitis.Methods:Eight-week-old female BALB/c mice were administered a 5%IMQ cream on both ears to produce psoriatic dermatitis.On day 3,K17 siRNA was mixed with an emulsion matrix and applied topically to the left ears of the mice after IMQ application every day for 7 days.The right ears of the mice were treated in parallel with negative control(NC)siRNA.Inflammation was evaluated by gross ear thickness,histopathology,the infiltration of inflammatory cells(CD3+T cells and neutrophils)using immunofluorescence,and the expression of cytokine production using real-time quantitative polymerase chain reaction.The obtained data were statistically evaluated by unpaired t-tests and a one-way analysis of variance.Results:The severity of IMQ-induced dermatitis on K17 siRNA-treated mice ears was significantly lower than that on NC siRNA-treated mice ears,as evidenced by the alleviated ear inflammation phenotype,including decreased ear thickness,infiltration of inflammatory cells(CD3+T cells and neutrophils),and inflammatory cytokine/chemokine expression levels(interleukin 17[IL-17],IL-22,IL-23,C-X-C motif chemokine ligand 1,and C-C motif chemokine ligand 20)(P<0.05 vs.the Blank or NC siRNA groups).Compared to the NC siRNA treatment,the K17 siRNA treatment resulted in increased K1 and K10 expression,which are characteristic of keratinocyte differentiation(vs.NC siRNA,K17 siRNA1 group:K1,t=4.782,P=0.0050;K10,t=3.365,P=0.0120;K17 siRNA2 group:K1,t=4.104,P=0.0093;K10,t=4.168,P=0.0042;siRNA Mix group:K1,t=3.065,P=0.0221;K10,t=10.83,P<0.0001),and decreased K16 expression,which is characteristic of keratinocyte proliferation(vs.NC siRNA,K17 siRNA1 group:t=4.156,P=0.0043;K17 siRNA2 group:t=2.834,P=0.0253;siRNA Mix group:t=2.734,P=0.0250).Conclusions:Inhibition of K17 expression by its specific siRNA significantly alleviated inflammation in mice with IMQ-induced psoriasis-like dermatitis.Thus,gene therapy targeting K17 may be a potential treatment approach for psoriasis.