消银解毒饮调控银屑病血热证肿瘤坏死因子-α的相关研究

目的 研究消银解毒饮对银屑病血热证外周血单一核细胞(PBMCs)分泌肿瘤坏死因子-α(TNF-α)的调控作用.方法 入选银屑病血热证患者抽取静脉血,分离PBMCs,并与人永生化角质形成细胞(HaCat细胞)混合接种培养48h,加入五种不同组别的动物药理血清,即蒸馏水空白组、消银解毒饮组、凉血方组、解毒方组、甲氨蝶吟阳性对照组,并于不同体积分数培养混合细胞48h,用四唑盐比色实脸(MTT)测定药物对HaCat细胞的细胞增殖率,双抗体夹心法测定药物对混合培养细胞上清TNF-α的影响.结果 消银解毒饮能有效的抑制HaCat细胞增殖(P<0.001),且效果优于拆方中药组(P<0.05);消银解毒饮能降低混合培养细胞分泌TNF-α(P<0.05),且与拆方两组比较疗效更显著(P<0.05).结论 消银解毒饮能有效的抑制角质细胞活化增殖,对银屑病血热证TNF-α有免疫调节作用,凉血类中药与解毒类中药协同作用治疗血热型银屑病效果可能更佳.     

消银解毒饮及拆方对角质形成细胞COLO-16增殖的调控作用

探讨消银解毒饮是否对角质形成细胞的增殖具有直接作用;以及消银解毒饮中凉血、解毒、祛风除湿等三组主要成分在治疗银屑病中所起的作用.本研究以角质形成细胞株COLO-16为研究对象;用四唑盐比色法观察了消银解毒饮及拆方后各组药物血清对COLO-16细胞增殖的影响.结果表明消银解毒组和凉血方组体外培养COLO-16细胞的存活率明显降低,而且存活率与含药血清浓度呈负相关.     

复方消银膏对小鼠银屑病样模型组织病理改变的研究

目的研究复方消银膏外用对BALB/c小鼠背部银屑病样模型组织病理的改变。方法 1分组:48只小鼠随机分为空白组、模型组、复方消银膏A组、复方消银膏B组、丙酸氯倍他索组及消银膏组。2造模:除空白组外其余各组鼠背部皮肤外用5%咪喹莫特,使其产生银屑病样模型。3治疗及评分:模型组作为对照,复方消银膏A组、复方消银膏B组、丙酸氯倍他索组及消银膏组分别给予相应药物治疗,根据8 d后光镜下各组局部皮肤组织病理变化进行BAKER评分,并进行统计学分析。结果空白组与模型组之间BAKER评分差异有统计学意义(P0.05),造模成功;与模型组相比,复方消银膏A组、复方消银膏B组、丙酸氯倍他索组的BAKER评分均降低,且差异均有统计学意义(P0.05);复方消银膏A组、复方消银膏B组与丙酸氯倍他索组相比差异有统计学意义(P0.05),而复方消银膏A组与复方消银膏B组之间差异无统计学意义(P0.05);消银膏组的BAKER评分低于模型组,而高于另外3个治疗组,且差异均有统计学意义(P0.05)。结论各治疗组均可以改善小鼠银屑病组织病理变化,丙酸氯倍他索组最明显,复方消银膏组次之,消银膏组最差,且复方消银膏A组与复方消银膏B组疗效无明显差异。     

消银解毒饮及拆方对角质形成细胞COLO-16凋亡的调控作用

本研究以角质形成细胞株COLO-16为研究对象,用流式细胞仪分析了消银解毒饮及其拆方后的凉血、解毒和祛风除湿等三组主要成份含药血清对其凋亡的诱导作用.结果表明消银解毒饮能诱导COLO-16细胞的凋亡,诱导角质形成细胞的凋亡可能是其治疗银屑病的机制之一.     

凉血解毒饮对血热型银屑病患者生活质量及P物质水平的影响

目的探讨自拟方凉血解毒饮对于血热型银屑病患者生活质量的影响及与P物质水平的关系。方法选择180例血热型银屑病患者,随机分为治疗组(凉血解毒饮组)和对照组(消银颗粒),每组90例,两组均外用卡泊三醇软膏,根据皮肤病生活质量指数(DLQI)、PASI评分方法判定疗效及对患者生活质量的影响程度,以双抗夹心ELISA方法来评定药物对P物质的影响及其相关性。结果治疗组疗效明显优于对照组,改善了患者的生活质量,且与P物质水平呈正相关。结论凉血解毒饮对血热型银屑病有一定的疗效,有效改善了患者的生活质量,与P物质水平的变化可能有关联。     

蜈蚣败毒饮对银屑病转基因模型小鼠外周血中Th17细胞表达影响的研究

目的探讨中药制剂蜈蚣败毒饮对银屑病转基模型小鼠外周血中Th17细胞表达的影响,初步从Th17细胞角度发现蜈蚣败毒饮的作用机制。方法采用随机对照的研究方法,选取银屑病转基因模型小鼠25只,将其分为5组,分别为空白对照组、甲氨蝶呤西药对照组、蜈蚣败毒饮低、中、高剂量实验组,每组5只转基因小鼠,经灌胃,含药PBMC制备,培养,染色等,采用流式细胞法检测转基因小鼠外周血PBMC中Th17细胞的表达情况。结果甲氨蝶呤组、蜈蚣败毒饮各剂量组与空白对照组比较,差异有统计学意义(P0.05);蜈蚣败毒饮低、中剂量组与甲氨蝶呤组比较,差异有统计学意义(P0.01);蜈蚣败毒饮高剂量组与甲氨蝶呤组比较,差异无统计学意义(P0.05)。结论蜈蚣败毒饮治疗银屑病的机制可能与降低外周血中Th17细胞的表达有关,其通过参与了IL-23-Th-17-IL-17的转导通路途径而发挥作用。     

蜈蚣败毒饮对银屑病小鼠滤泡辅助性T细胞上下游因子的干预作用

目的 观察蜈蚣败毒饮对滤泡辅助性T(Tfh)细胞上、下游因子白细胞介素-6(IL-6)、核转录基因(Bcl-6)、可诱导共刺激分子(ICOS)以及肿瘤坏死因子相关激活蛋白(CD40L)表达的干预,探讨其对银屑病的治疗机制。方法 将C57BL/6小鼠30只随机分6组,每组5只,并依次命名为空白组、模型组、西药对照组、蜈蚣败毒饮低、中、高剂量组。咪喹莫特诱导建立银屑病小鼠模型。建模完成后空白组和模型组小鼠给予生理盐水灌胃,各给药组小鼠药物灌胃处理。评估治疗前后各组小鼠的银屑病皮损面积和严重程度指数(MPASI),观察皮损微观病理改变情况、qRT-PCR法检测IL-6、Bcl-6、ICOS、CD40L mRNA水平,Western blot法测定IL-6、Bcl-6、ICOS、CD40L蛋白含量,最后对皮损检测指标与皮损改善程度进行相关性分析,用以评价皮损与该通路的关联。结果 药物处理后蜈蚣败毒饮各剂量组的小鼠MAPSI分值均出现下降(P<0.01),△MPASI均表现为升高(P<0.01),且蜈蚣败毒饮高剂量组与西药对照组表现相当,差异无统计学意义(P>0.05);皮损H...     

消银解毒饮对银屑病患者鳞屑白介素8的影响

为了研究中药治疗银屑病的机理，我们在 1999年 2月用双抗体夹心 ELISA法对银屑病患者服用消银解毒饮前后鳞屑中白介素 8(IL－ 8)含量进行测定。 一、资料和方法 检测对象： 1998年 9月至 1999年 2月北京中医药大学东直门医院皮肤科门诊就诊的寻常型进行期银屑病患者。观察期间，患者口服消银解毒饮，每次 50 mL，日 2次 ,皮损局部外搽凡士林软膏，就诊前 1 d皮损处不使用外用药膏，用药前及用药 4周后，刮取鳞屑，收集 20例患者的鳞屑 30份，其中服用消银解毒饮治疗前的鳞屑 20份，用药 4周后的鳞屑 10份（另有 10份疗后的鳞屑，因量…     

窄谱中波紫外线联合消银颗粒治疗银屑病临床观察

目的:观察窄谱中波紫外线(NB-UVB)联合消银颗粒治疗寻常性银屑病能否明显降低复发率.方法:将122 例寻常性银屑病患者随机分为两组.治疗组:NB-UVB照射联合消银颗粒内服.对照组:只行NB-UVB 照射.每周照射3次,疗程为8 周.试验结束后依据患者治疗前后症状、体征进行评价,随访3 个月,评估治疗后银屑病复发率.结果:治疗组有效率为70.97%,3个月后复发率6.82%;对照组有效率为61.67%,3 个月后复发率37.84%.两组有效率差异无统计学意义(P ＞ 0.05),复发率差异有统计学意义(P ＜ 0.05).结论:NB-UVB 联合消银颗粒治疗寻常性银屑病疗效好,不良反应少,能明显降低复发率.     

S100A10蛋白在银屑病皮损的表达及对小鼠银屑病样皮炎模型的影响

目的探讨S100A10蛋白在银屑病皮损的表达以及对咪喹莫特诱导小鼠银屑病样皮炎的影响。方法 2020年1月至2022年6月在新乡医学院第一附属医院皮肤科通过外科手术收集28例银屑病患者皮损, 同期于普通外科及眼科收集18例健康人皮肤作为对照组。通过免疫组化检测S100A10蛋白在银屑病患者皮损和健康对照皮肤的表达。分别选取10只野生型(WT)C57BL/6J雌性小鼠及10只S100A10-/- C57BL/6J雌性小鼠, 建立咪喹莫特(IMQ)诱导的银屑病样皮炎小鼠模型, 采用随机数字表法分为基因敲除(KO)/IMQ组、WT/IMQ组、KO/凡士林(VAS)组及WT/VAS组, 每组5只, 背部剃毛后, KO/IMQ及WT/IMQ组每日涂抹IMQ乳膏(62.5 mg)于背侧皮肤建立银屑病样皮炎小鼠模型, KO/VAS及WT/VAS组每日涂抹凡士林乳膏(62.5 mg), 连续7 d, 每日观察小鼠背部皮损情况, 于第7天采用颈椎脱臼法处死, 切取背部皮肤组织。每日通过银屑病皮损面积及严重程度指数(PASI)评价IMQ诱导的小鼠银屑病样皮炎, HE染色观察皮损病理表现, 免疫组化染色检测...     

Stat3在进行期寻常型银屑病皮损表皮中的表达

目的观察信号转导和转录激活因子3(Stat3)在银屑病皮损中的表达情况。方法应用免疫组化法检测正常组织、进行期寻常型银屑病皮损及非皮损表皮中Stat3蛋白表达。结果正常表皮细胞中Stat3低水平表达于胞浆中;寻常型银屑病进行期皮损表皮中Stat3表达于多数细胞的胞浆和胞核中;非皮损处表皮中Stat3表达于部分细胞的胞浆和(或)胞核中。皮损处Stat3评分与PASI呈正相关。结论Stat3在进行期寻常型银屑病表皮中有异常表达。     

(8) Comparison of psoriasis treated with cyclosporin alone or cyclosporin and clobetasol propionate

Cyclosporin A (CyA) in the low dosage of 3 mg/kg/day has been shown to clear psoriasis; however, the disease relapses soon after withdrawal of the drug. Immunological studies have demonstrated persistence of activated T helper (TH) cells within the epidermis after clearance of psoriatic plaques with cyclosporin but disappearance if clearance is achieved by the use of topical clobetasol propionate (CP). If the relapse is a result of resumption of function by epidermal-activated TH cells then combination therapy with CP and CyA should delay the rate of relapse.
Six patients with chronic plaque psoriasis were treated with CyA at a dose of 3 mg/kg/day for a period of 6 weeks; during the first 2 weeks CP was applied twice daily to the psoriatic plaques. A second group of six psoriatic patients was treated with cyclosporin alone. The psoriasis was scored at weekly intervals during treatment using the Psoriasis Area and Severity Index (PASI), and for 4 weeks after cessation of treatment.
Using CyA alone, the mean time taken to achieve 80% reduction in PASI score was 7·3 ± 2·4 weeks compared to 4·2 ± 2·1 weeks with CyA and CP ( P < 0·05). Four weeks after withdrawal of treatment the average percentage deterioration in PASI score was 29·2 in the CyA group and 17·8 in the CyA + CP group. This difference was not significant.
Thus, the addition of CP to CyA in the management of psoriasis cleared psoriasis faster than CyA alone but did not slow the relapse rate. It would seem that the initiating factor responsible for attracting TH cells into the epidermis is not cleared by either treatment.     

解毒化淤汤治疗银屑病的临床和实验研究

目的 研究解毒化淤汤治疗银屑病的临床疗效和作用机制.方法应用解毒化淤汤对50例寻常性银屑病患者进行临床治疗观察,并利用小鼠尾部作银屑病动物模型,以解毒化淤汤灌胃,观察其对小鼠尾部颗粒细胞形成的影响.结果解毒化淤汤治疗银屑病的临床治愈率为42.00%,总有效率为90.00%.动物实验表明解毒化淤汤能明显地增加小鼠尾部鳞片表皮的颗粒层,与对照组比较差异有显著性.结论解毒化淤汤是治疗寻常性银屑病的有效方剂,其作用机制可能与调节自身免疫功能、抑制表皮细胞的增殖有关。     

The relationship between severity of the disease and circulating nucleosomes in psoriasis patients

Psoriasis was initially considered to represent a disease of abnormal epidermal keratinocyte proliferation. Proliferation of keratinocytes is restricted by apoptotic cell death to maintain a constant thickness of epidermis. Nucleosomes are mainly released by apoptotic cells. Tumor necrosis factor-α (TNF-α) is an important factor affecting the apoptosis. In the present study, the relationship between TNF-α, nucleosome and the Psoriasis Area and Severity Index (PASI) score was investigated. The patients were divided according to PASI score into three groups (mild, moderately, severe). Serum TNF-α and nucleosome levels were measured using Enzyme-linked immunosorbent assay (ELISA) method. Our findings show a correct relationship between PASI scores and TNF-α and an inverse relationship between nucleosome and PASI score. According to the results obtained from the study, we believe that serum nucleosome levels can be used as a new indicator in follow-up of patients with psoriasis and monitoring of the effectiveness of drugs which used in the treatment of psoriasis.     

郁金银屑片对Balb/c裸鼠银屑病模型的实验研究

目的观察郁金银屑片对Balb/c裸鼠银屑病模型皮损组织中角质细胞生长因子(Keratinocyte growth factor,KGF)和双调蛋白(Amphiregulin,AREG)及两者基因表达的影响。方法将36只Balb/c裸鼠随机分为空白对照组、模型组和郁金银屑组,除空白对照组外,另2组裸鼠均用10%普萘洛尔涂于背部皮肤,2次/d,连续1周的方法造银屑病模型。成模后郁金银屑组按0.25 g/kg剂量灌郁金银屑片混悬液治疗2周,空白对照组和模型组灌等量生理盐水。对裸鼠背部皮损进行银屑病皮损面积及严重程度指数(PASI)评分;以酶联免疫吸附法(ELISA)定量检测皮损组织均浆液KGF和AREG的蛋白量,实时定量反转录聚合酶链反应(RT-PCR)方法检测KGF m RNA和AREG m RNA的表达。结果模型组和郁金银屑组造模后PASI评分、匀浆液中KGF和AREG明显升高,KGF m RNA和AREG m RNA高表达,与空白对照组比较,P0.05。在治疗2周后,模型组与同组治疗前比较,P0.05;郁金银屑组PASI评分、匀浆液中KGF和AREG降低,KGF m RNA和AREG m RNA低表达,与同组治疗前和模型组治疗后比较,P0.05,差异有统计学意义。结论郁金银屑片治疗银屑病机制与降低表皮组织匀浆液中KGF和AREG、下调两者基因表达密切相关。     

Leptin induces secretion of pro‐inflammatory cytokines by human keratinocytes in vitro – a possible reason for increased severity of psoriasis in patients with a high body mass index

Investigations about prevalence of obesity in psoriasis patients are increased nowadays. Higher serum levels of leptin in patients with psoriasis who are overweight or obese suggest that leptin may serve as a molecular link between psoriasis and metabolic comorbidities. However, the pathological functions of leptin in psoriasis are not clearly understood. We investigated the influence of being overweight or obese on the risk of psoriasis, and the relationship between serum leptin levels and the severity of psoriasis in Chinese Han patients. We also investigated biological effects of leptin on the proliferation and secretion of pro‐inflammatory cytokines by human keratinocytes in vitro. Obesity was a significant risk factor for psoriasis in the Chinese Han population; however, we did not observe a significant correlation between Psoriasis Area and Severity Index (PASI) and body mass index (BMI). We observed a positive correlation between the serum leptin level and PASI in overweight and obese male patients with psoriasis. Strong leptin immunoreactivity was detected in the epidermis of psoriatic lesions, particularly in keratinocytes. Leptin significantly increased the proliferation and secretion of pro‐inflammatory cytokines by keratinocytes in vitro. In conclusion, this study suggests leptin as a novel molecular link between psoriasis and obesity, which may help to explain the more server conditions of psoriasis in patients with obesity.     

转录因子CCAAT/增强子结合蛋白α在银屑病皮损中的表达及意义

目的 探讨转录因子CCAAT/增强子结合蛋白α（C/EBP-α）在寻常性银屑病皮损中的表达及与角质形成细胞异常增殖及皮损严重程度间的相关性。 方法 用免疫组化二步法及Western印迹检测30例寻常性银屑病患者皮损和30例健康对照皮肤的表皮中C/EBP-α。免疫组化法检测Ki-67的表达,并根据Ki-67阳性表达数量计算增殖指数,Pearson相关分析法分析寻常性银屑病皮损中C/EBP-α的表达水平与角质形成细胞Ki-67增殖指数及银屑病皮损面积和严重程度指数（PASI评分）间的相关性。 结果 C/EBP-α主要在角质形成细胞胞质中表达,寻常性银屑病皮损中C/EBP-α的表达较健康对照皮肤明显下调（t = 7.82,P ＜ 0.05）。Ki-67主要在角质形成细胞胞核中表达,寻常性银屑病皮损中角质形成细胞的增殖指数明显高于健康对照皮肤（t = 4.54,P ＜ 0.05）。经Pearson相关分析,寻常性银屑病皮损中C/EBP-α表达水平与增殖指数呈负相关,与PASI评分呈负相关。Western印迹结果亦显示,C/EBP-α在银屑病皮损处的表达量显著下调。 结论 C/EBP-α在寻常性银屑病皮损中表达下调,可能参与了寻常性银屑病的发病过程。     

养血解毒汤对小鼠银屑病样皮损IL-36表达的影响

 目的: 探讨养血解毒汤对小鼠银屑病样模型皮损IL-36表达的影响。方法：将18只小鼠随机分为对照组、模型组及实验组,每组6只,对各组小鼠皮损进行PASI评分;通过RT-qPCR比较各组小鼠皮损中IL-36α、IL-36β、IL-36γ与IL-36Ra mRNA表达差异;通过Western blot及免疫组化比较各组小鼠脱毛区域皮肤IL-36β的蛋白表达。结果：实验组小鼠皮损PASI评分（1.17±0.37）明显低于模型组（7.83±0.69）,差异有统计学意义（t=19.07, P<0.05）。 实验组IL-36β的mRNA表达（1.85±0.28）较模型组（4.85±0.42）明显降低（t=8.01,P<0.01）,而IL-36α、IL-36γ及IL-36Ra的mRNA水平无明显改变。IL-36β在模型组中大量表达于颗粒层以及棘层中上部的细胞胞核和胞浆,且真皮层也有少量分布;在实验组中少量表达于真皮层,少量或无表达于表皮层。与模型组（3.27±0.32）相比,实验组小鼠皮损中IL-36β蛋白表达（2.50±0.24）明显降低（t=3.31,P<0.05）。结论：养血解毒汤通过下调IL-36β表达水平,有效改善小鼠银屑病样皮损严重程度,降低皮损PASI评分。     

Normal keratinocytes express Toll-like receptors (TLRs) 1, 2 and 5: modulation of TLR expression in chronic plaque psoriasis

BACKGROUND: Toll-like receptors (TLRs) are part of the innate immune system involved in the response to microbial pathogens. TLR2 recognizes various ligands expressed by Gram-positive bacteria, while TLR3, TLR4 and TLR5 are specific for double-stranded RNA, Gram-negative lipopolysaccharides and bacterial flagellin, respectively. OBJECTIVES: To determine, firstly, whether epidermal keratinocytes of normal skin express TLRs and, secondly, whether modulation of TLR expression occurs in psoriasis, an inflammatory skin disease associated with certain microorganisms such as streptococci, staphylococci and yeasts. METHODS: Eight samples of normal, and 15 samples of lesional and nonlesional psoriatic skin were stained with polyclonal antibodies specific for TLR1-5 using an avidin-biotin-peroxidase technique. RESULTS: Epidermal keratinocytes in normal skin constitutively expressed TLR1, TLR2 and TLR5, while TLR3 and TLR4 were, in most cases, barely detectable. Cytoplasmic TLR1 and TLR2 were expressed throughout the epidermis, with higher staining of the latter on basal keratinocytes, while TLR5 expression was concentrated in the basal layer. In contrast, in lesional epidermis from patients with psoriasis, TLR2 was more highly expressed on the keratinocytes of the upper epidermis than on the basal layer, while TLR5 was downregulated in basal keratinocytes compared with corresponding nonlesional psoriatic epidermis. In addition, nuclear TLR1 staining was observed in the upper layers of both nonlesional and lesional psoriatic epidermis, but not in that of normal skin. CONCLUSIONS: These findings suggest that TLRs expressed by epidermal keratinocytes constitute part of the innate immune system of the skin. The relevance of altered keratinocyte TLR expression in psoriasis remains to be determined.     

Pilot trial: Pioglitazone versus placebo in patients with plaque psoriasis (the P6)

BACKGROUND: Disordered differentiation and hyperproliferation of keratinocytes with inflammation are the hallmarks of psoriasis. Ligand activation of peroxisome proliferator receptor-gamma (a class of nuclear receptors) by thiazolidinediones can normalize the histologic features of psoriasis. METHOD: In a 10-week, double-blind, randomized, placebo-controlled, parallel-group study, 70 patients with moderate to severe psoriasis received one of the following treatments: pioglitazone 15 mg, pioglitazone 30 mg or placebo. Efficacy was evaluated by observing the change in the psoriasis area and severity index (PASI) after 10 weeks of treatment. RESULTS: There was a dose-dependent improvement in psoriasis. Median PASI scores at the end of 10 weeks were significantly reduced in the pioglitazone treatment groups as compared to the placebo-treated group. The psoriasis lesions cleared in more than 40% of patients treated with pioglitazone as compared to 12.5% of those with placebo. The percentage reduction in mean PASI scores was 21.6%, 41.1% and 47.5% in the placebo, pioglitazone 15 mg, and 30 mg groups, respectively. No serious adverse events were detected. CONCLUSION: This is the first report from a controlled trial demonstrating that pioglitazone could be considered as an efficacious and safe agent for the treatment of plaque psoriasis. The optimum dose and duration of pioglitazone therapy remain to be determined.