白癜风患者血清过氧化氢、过氧化氢酶和谷胱甘肽过氧化物酶的检测

目的探讨体内氧化物和抗氧化物与白癜风发病的关系。方法分别检测40例白癜风患者和10例健康对照者的血清过氧化氢(H2O2)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-PX)。结果白癜风患者CAT浓度(9.31±6.52)U/mL明显低于对照组(33.05±9.47)U/mL,其进展期CAT浓度(7.3±6.01)U/mL明显低于稳定期(13.05±6.11)U/mL,差异均有统计学意义(P均<0.01);而白癜风患者H2O2和GSH-PX水平与对照组比较,差异无统计学意义(P>0.05)。结论白癜风的发生可能与血清氧化物—抗氧化物水平的变化有一定的相关性。     

白癜风患者血浆中氧化与抗氧化相关指标的检测

目的通过对白癜风患者及健康对照者血浆中氧化与抗氧化相关指标——超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、维生素E(VitE)及一氧化氮(NO)表达水平的测定,探讨氧化应激在白癜风发病机制及疾病发展中的作用和意义。方法选择60例白癜风患者(患者组)和40名健康志愿者(健康对照组)为研究对象,化学法检测血浆中SOD,CAT,GSH-Px的活性及MDA,VE和NO的含量。结果患者组血浆中MDA含量及SOD活性明显高于健康对照组(P<0.01),而GSH-Px活性明显低于健康对照组(P<0.01);进展期白癜风患者血浆中MDA的含量及SOD的活性明显高于稳定期白癜风组(P<0.01),而GSH-Px活性明显低于稳定期白癜风患者(P<0.01);随MDA含量增加,GSH-Px活性逐渐降低,呈显著负相关关系(r=-0.337,P<0.01),而SOD活性逐渐升高,呈显著正相关关系(r=0.347,P<0.01);而在进展期和稳定期白癜风患者血浆中CAT,VE和NO的含量与健康对照组相比差异无统计学意义(P>0.05)。结论白癜风患者血浆中存在氧化-抗氧化失衡,白癜风的发病及病情活动与氧化应激可能相关。     

白癜风患者外周血Th17细胞相关因子的表达

为检测Th17细胞相关因子IL-17A、IL-17F、IL-22、IL-23与白癜风的关系,随机抽取30例白癜风患者,进行期14例,稳定期16例,正常对照组30例,采用SYBR Green I实时荧光定量RT-PCR技术,检测外周血单个核细胞(PBMC)IL-17A mRNA,IL-17F mRNA,IL-22 mRNA的相对表达量.采用双抗体夹心ELISA法检测血浆IL-23的含量.进行期白癜风患者PBMC IL-17A mRNA的相对表达量和稳定期白癜风患者与正常对照组相比明显增高,有统计学差异(P=0.00;P=0.001).进行期和稳定期白癜风患者与正常对照组相比,PBMC IL-17F mRNA,IL-22 mRNA的相对表达水平无显著性差异.白癜风进行期与稳定期患者血浆IL-23浓度与正常对照组相比明显增高,有统计学差异(P=0.01;P=0.00).白癜风患者PBMC IL-17A mRNA的相对表达量和血浆IL-23浓度高表达,提示Th17细胞可能参与白癜风的发病.     

白癜风白斑和复色斑氧化一抗氧化物水平比较分析

目的:确定体内氧化一抗氧化状态与白癜风发病的关系.方法:对24例白癜风患者白斑、非白斑(正常皮肤)、11例白癜风患者复色斑和10例健康对照者的皮肤负压吸疱,抽取组织液,检测过氧化氢(H2O2)、过氧化氢酶(CAT)与谷胱甘肽过氧化物酶(GSH-PX)水平.结果:白斑组H2O2含量为(53.97±19.03)mmol/L,明显高于健康对照组的(28.98±22.81)mmol/L、非白斑的(33.41±21.53)mmol/L和复色斑的(23.82±16.28)mmol/L(P＜0.01);白斑组CAT含量为(17.34±11.05)U/mL,明显低于健康组的(41.29±16.57)U/mL、非白斑的(39.46±19.88)U/mL和复色斑的(33.66±10.51)U/mL(P分别＜0.01,＜0.05,P＜0.01).H2O2和CAT含量在健康组、非白斑和复色斑之间无显著性差异(P＞0.05);复色斑中的GSH-PX浓度高于健康组(P＜0.05),其余各组相互比较GSH-PX水平无差异(P＞0.05).结论:白癜风发病可能与体内H2O2增高、CAT降低密切相关,而与GSH-PX无关.     

白癜风患者白介素-17和转化生长因子-β水平的检测

目的 探讨白介素-17(IL-17)和转化生长因子-B(TGF-β)与白癜风发病之间的关系.方法 双抗体夹心酶联免疫吸附试验( ELISA)检测120例白癜风患者和60例健康对照血清中IL-17和TGF-β水平.分析两种因子与白癜风患者性别、病期、病程、白斑面积、家族史的关系.结果 进展期白癜风患者血清IL-17水平显著高于健康对照组和稳定期患者(P值均＜0.05),随着白癜风患者白斑面积增大,IL-17水平逐渐升高(x2=12.656,P＜ 0.05);而进展期白癜风患者血清TGF-β水平也高于健康对照组和稳定期患者,但差异无统计学意义(P值均＞ 0.05).结论 白癜风患者血清IL-17和TGF-β水平与病情有一定关系,两者可能在白癜风发病中发挥作用.     

肿瘤坏死因子-α及其受体在白癜风患者皮损中的表达

目的探讨肿瘤坏死因子-α(TNF-α)及其受体(TNFR1)与白癜风发病的关系。方法采用SABC免疫组化法对18例进展期及20例稳定期白癜风患者皮损处TNF-α及TNFR1进行检测,并以20例正常人作对照。结果寻常型进展期白癜风皮损中TNF-α的表达较寻常型稳定期组显著增加(P<0.01),寻常型稳定期白癜风皮损中TNF-α的表达较正常对照组显著增加(P<0.001)。患者组与对照组TNFR1无显著性差异(P>0.05)。结论TNF-α可能参与寻常型白癜风发病的某一环节。     

白癜风患者血清硒浓度检测分析

探讨微量元素硒在白癜风发病机制中的作用.检测分析36例白癜风患者和健康者的血清硒浓度.白癜风患者、进展期患者、稳定期患者及健康者血清硒浓度,分别为(121.9±46.16)ng/mL,(121.08±44.83)ng/mL,(122.72±48.74)ng/mL,(129.27±23.67)ng/mL,它们之间均无显著性差异(P＞0.05).提示微量元素硒与白癜风的发病机制可能无关系.     

白癜风患者血清可溶性细胞间黏附分子-1的检测

目的检测白癜风患者血清可溶性细胞间黏附分子-1(sICAM-1)的水平,并探讨其与白癜风活动性关系.方法采用酶联免疫吸附试验(ELISA)法检测40名健康人和40例白癜风患者血清sICAM-1水平.结果①白癜风患者血清sICAM-1浓度明显增高,与正常对照组比较有显著性差异(P＜0.001).②进展期白癜风患者血清sICAM-1浓度显著高于稳定期(P＜0.001).③稳定期白癜风患者血清sICAM-1浓度与正常对照组无显著性差异(P＞0.05).结论 sICAM-1与白癜风活动程度有关.     

进展期白癜风患者治疗后血清中sICAM-1、IFN-γ的变化

目的探讨进展期白癜风患者治疗后血清中可溶性细胞间黏附分子-1(s ICAM-1)、干扰素-γ(IFN-γ)水平的变化及临床意义。方法收集33例进展期白癜风患者作为治疗组,采用酶联免疫吸附试验(ELISA)方法检测患者血清中sICAM-1、IFN-γ水平,并与20例稳定期白癜风患者及20例健康人对照;33例进展期白癜风患者均给予综合治疗,总疗程为16周,疗程结束后评定疗效并再次检测血清sICAM-1、IFN-γ水平。结果进展期白癜风患者血清s ICAM-1、IFN-γ平均水平明显高于稳定期白癜风患者及正常人,差异有统计学意义;稳定期白癜风患者血清sICAM-1、IFN-γ平均水平与正常人比较差异无统计学意义;治疗后进展期白癜风患者血清sICAM-1、IFN-γ平均水平较治疗前明显降低,差异有统计学意义;治疗有效组进展期白癜风患者治疗后血清sICAM-1、IFN-γ降低程度高于治疗无效组,差异有统计学意义。结论白癜风患者血清s ICAM-1、IFN-γ可反映疾病活动性,其水平变化可能与进展期白癜风的转归及临床疗效相关。     

白癜风患者白介素-17和转化生长因子-β水平的检测北大核心CSCD

目的 探讨白介素-17（IL-17）和 转化生长因子-β（TGF-β）与白癜风发病之间的关系。方法 双抗体夹心酶联免疫吸附试验（ELISA）检测120例白癜风患者和60例健康对照血清中IL-17和TGF-β水平。分析两种因子与白癜风患者性别、病期、病程、白斑面积、家族史的关系。 结果 进展期白癜风患者血清IL-17水平显著高于健康对照组和稳定期患者（P值均 〈 0.05）,随着白癜风患者白斑面积增大,IL-17水平逐渐升高（χ2 = 12.656,P 〈 0.05）;而进展期白癜风患者血清TGF-β水平也高于健康对照组和稳定期患者,但差异无统计学意义（P值均 〉 0.05）。 结论 白癜风患者血清IL-17和TGF-β水平与病情有一定关系,两者可能在白癜风发病中发挥作用。     

Characterization of glutathione S-transferase in cultured human keratinocytes

The glutathione S-transferase activity and isozymic composition of cultured human keratinocytes were characterized. Keratinocytes were grown in culture and harvested at different stages of differentiation. Glutathione S-transferase activity was found in the soluble cell fraction but not in the microsomal cell fraction. The glutathione S-transferase specific activity of the soluble cell fraction was found to increase as the keratinocytes differentiated in culture. All of the enzymatic activity was found to reside with a single isozymic form that was concluded to be the pi form of the enzyme based on substrate specificity, sensitivity to inhibitors, molecular weight, and reactivity towards antibodies raised to alpha, mu, and pi forms of the enzyme. It is concluded that all of the isozymic forms of glutathione S-transferase noted in whole skin, with the exception of pi, are of extra-keratinocyte origin.     

Expression of glutathione S-transferase-pi in malignant skin tumors.

GST-pi has been known to be markedly increased in human (pre) neoplasms of several organs. In this paper, the significance of immunohistochemical detection of GST-pi in human malignant tumors of the skin was studied. In specimens from 40 patients with various skin cancers, malignant melanoma, Paget's disease and undifferentiated squamous cell carcinoma showed strong reactivity in GST-pi staining. The reactions were negative or weak in Bowen's disease, basal cell epithelioma and solar keratosis. In normal melanocytes, eccrine, apocrine, and breast gland cells stained positively but not in keratinocytes, sebaceus gland and fibroblasts. While immunohistochemical detection of GST-pi in the skin was not specific for malignancies, it contributed to aid the distinction of squamous cell carcinoma from other keratinocytic tumors. GST-pi might provide potentially useful information on chemosensitivity of skin cancer, and might serve as a biomarker of disease activity.     

Modulation of glutathione level in cultured human melanoma cells

The effects of buthionine sulphoximine (BSO) treatment on cellular glutathione (GSH) content and on the cytotoxic action of menadione were investigated in cultured IGRI human melanoma cells. Addition of BSO (10(-8)-0.5 X 10(-3) M) to the cultures resulted in a dose- and time-dependent depletion of cellular GSH. BSO (10(-5) and 10(-6) M) did not influence cell multiplication up to 48 h, as determined by trypan blue staining. Menadione (3 X 10(-5) M) treatment decreased the cellular GSH concentration and also reduced cell number after a 24 h exposure. Its cytotoxicity was increased by BSO (10(-5), 10(-6) M), though the potentiating effect was moderate.     

胎盘型谷胱甘肽转换酶在皮肤角朊细胞肿瘤中的表达

为了探讨谷胱甘肽转换酶π（ＧＳＴπ）与皮肤角朊细胞肿瘤发生的关系，采用免疫组化技术检测了４５例皮肤角朊细胞肿瘤和５例正常皮肤。结果∶ＧＳＴπ在正常皮肤主要表达于颗粒细胞层、大部分棘细胞层和外毛根鞘、小汗腺。在日光性角化病、Ｂｏｗｅｎ病、基底鳞状细胞癌和鳞状细胞癌中胞浆和部分胞核ＧＳＴπ阳性，基底细胞癌胞浆弱阳性，部分脂溢性角化症和角化棘皮瘤胞浆呈弱阳性。本研究提示ＧＳＴπ主要存在于表皮上层，并在皮肤角朊细胞肿瘤发生中起一定作用。     

E6* oncoprotein expression of human papillomavirus type-16 determines different ultraviolet sensitivity related to glutathione and glutathione peroxidase antioxidant defence

Clinical observations of non-melanoma skin cancer in immunocompromised patients, such as organ transplant recipients, suggest co-operative effects of human papillomavirus (HPV) and ultraviolet (UV) radiation. The aim of the present study is to evaluate UV sensitivity and DNA damage formation according to antioxidant status in HPV16-infected keratinocytes. We used SKv cell lines, infected with HPV16 and well characterized for their proliferative and tumorigenic capacities. We showed that SKv cell lines presented various E6* (a truncated form of E6) RNA levels. We demonstrated that the higher oncoprotein RNA expression level was associated with a higher resistance to solar-simulated radiation, more specifically to UVB radiation and to hydrogen peroxide. Moreover, this high resistance was associated with a low oxidative DNA damage formation after UV radiation and was related to high glutathione content and glutathione peroxidase activities. Therefore, the results of our study suggest that E6* levels could modulate the glutathione/glutathione peroxidase pathway providing a mechanism to protect HPV-infected keratinocytes against an environmental oxidative stress, such as UV radiation.     

Decrease in glutathione may be involved in pathogenesis of acne vulgaris

Background Some past studies reported that oxidative stress components such as reactive oxygen species (ROS) or lipid peroxide (LPO) are involved in the pathogenesis and progression of acne vulgaris. In this study, we hypothesized that the pathogenesis of acne vulgaris may depend on the differences in antioxidative activity among antioxidants in our body. We collected samples of stratum corneum from acne patients and healthy subjects and compared the quantity of gluthathione (GSH), one of many antioxidative components in our body, for comparison. Methods Samples of stratum corneum were collected from facial acne‐involved lesion, facial uninvolved area, and the medial side of the upper arm in acne vulgaris patients. Similarly, samples were collected from a facial uninvolved area and the medial side of the upper arm in healthy subjects. The quantity of GSH was measured in each area. In vitro effects of alpha‐melanocyte stimulating hormone (α‐MSH) on GSH synthesis‐related gene were also examined. Results The quantity of GSH in stratum corneum from each area was significantly lower in acne vulgaris patients than that of healthy subjects. There was no significant difference in quantity of GSH between the acne‐involved lesion and uninvolved area in acne patients. In vitro studies showed that the expression level of Glutamate‐cysteine ligase catalytic subunit (GCLC), one of the GSH synthesis‐related genes, was significantly decreased by the additional use of α‐MSH. Conclusions We conclude that a decline in antioxidative activity led by a decrease in GSH quantity may play an important role in pathogenesis of acne vulgaris. The use of α‐MSH may further decrease the GSH level.     

Serum selenium levels and blood glutathione peroxidase activities in vitiligo

It has recently been shown that patients with vitiligo can accumulate epidermal hydrogen peroxide (H2O2) in association with low catalase levels. This study examined serum selenium levels and blood glutathione peroxidase activities in 61 patients and controls. The results showed high serum selenium levels in 56% of the patients. As at least one isoform of glutathione peroxidase requires selenium for its activity, enzyme activities were also evaluated. The overall results were not significantly different compared with controls, but further age-related analysis of the data indicated significantly lower activities in patients up to 46 years. As glutathione peroxidase can also efficiently degrade H2O2, the results of this study could indicate an additional impaired H2O2 metabolism in vitiligo.