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1.
研究抗组胺药盐酸西替利嗪对表皮角质形成细胞系HaCaT细胞和真皮成纤维细胞P物质受体表达以及对P物质诱导两种细胞表达IFN-γ、IL-1β、IL-6及IL-8的影响。采用流式细胞术和Western blotting检测分析西替利嗪对两种皮肤细胞P物质受体表达的影响;以P物质刺激HaCaT细胞和真皮成纤维细胞,加入不同剂量的西替利嗪共孵育,采用ELISA方法测定西替利嗪对IFN-γ、IL-1β、IL-6及IL-8分泌的影响。结果表明,西替利嗪显著抑制HaCaT细胞和真皮成纤维细胞P物质受体的表达;P物质刺激可以显著增加HaCaT细胞和真皮成纤维细胞IFN-γ、IL-1β、IL-8的分泌量;1~100 μmol·L-1的西替利嗪均可抑制皮肤细胞IL-1β、IL-8的分泌,但对IFN-γ的分泌无明显影响。P物质和西替利嗪对两种皮肤细胞IL-6的产生均无显著影响。  相似文献   

2.
目的研究地塞米松、他克莫司(FK-506)和CX-659s对胸腺活化调节趋化因子(TARC/CCL17)产生的调控作用。方法运用ELISA方法分别研究了3种药物对人角质形成细胞株——HaCd细胞和人成纤维细胞株——NG1RGB细胞的TARC表达作用的影响。结果3种药物对于HaCaT细胞中TARC的表达,10^-9~10^-5mol/L地塞米松显示明显的抑制作用;对于IFN-γ刺激下HaCaT细胞产生TARC水平,10^-12~10^-6mol/L地塞米松、10^-12~10^-6mol/L的FK-506均表现为明显的抑制作用(P〈0.05或P〈0.01);CX-659s显示明显抑制NGlRGB细胞中TARC水平,且具有浓度依赖性(P〈0.01)。3种免疫抑制剂分别在不同条件下抑制皮肤角质形成细胞和/或成纤维细胞的TARC表达。结论这些药物明显抑制TARC在皮肤角质形成细胞和成纤维细胞的表达,因此认为角质形成细胞和成纤维细胞可能在病理状态下通过促进CCR4+T细胞向炎症部位的游走,参与免疫调控作用。  相似文献   

3.
目的 考察IL-17与TNF-α共作用对皮肤角质形成细胞系HaCaT细胞IL-17R、p-p38 MAPK表达的影响,及其对HaCaT细胞分泌IL-6、IL-8、MIP-3α等炎症因子的影响。方法 采用RT-PCR方法检测IL-17与TNF-α共作用对皮肤角质形成细胞IL-17R mRNA表达的影响;采用western blot方法检测IL-17与TNF-α共作用对HaCaT细胞表达p-p38 MAPK的影响;采用ELISA法检测IL-17与TNF-a共作用对HaCaT细胞分泌炎症因子IL-6、IL-8及MIP-3α的影响。结果 IL-17与TNF-α共作用可上调HaCaT细胞表达IL-17R和p-p38 MAPK,以及上调HaCaT细胞IL-6、IL-8、MIP-3α等炎症因子的分泌。结论 IL-17与TNF-α共作用可通过上调皮肤角质形成细胞IL-17R的表达,促进p38 MAPK的磷酸化过程而具有明显致炎作用。  相似文献   

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目的考察西替利嗪(CET)对皮肤细胞炎症模型中单核趋化蛋白-1(MCP-1)表达的干预作用。方法组胺(HIS)和IFN-γ刺激真皮成纤维细胞(DF)和人角质形成细胞株HaCaT细胞,采用RT-PCR和ELISA法考察两种细胞MCP-1 mRNA和蛋白表达水平。结果与对照组比较,HIS(10 μmol·L-1)和IFN-γ(20 ng·mL-1)组显著上调两种细胞(DF和HaCaT) MCP-1 mRNA表达,同时分别使DF的MCP-1蛋白分泌量增加3.5倍和8.4倍,对HaCaT细胞也有相似的影响趋势; CET (1和10 μmol·L-1) 显著地抑制了它们对细胞MCP-1蛋白产生的增强作用。结论CET可能通过抑制MCP-1的表达而发挥抗皮肤炎症作用。  相似文献   

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目的体外构建适用于经皮给药研究的组织工程皮肤。方法以表皮角质形成细胞系HaCaT细胞及人真皮成纤维细胞为细胞来源,用I型牛胶原蛋白为真皮基质包埋成纤维细胞,其上接种HaCaT细胞,采用气-液界面方式培养,观察不同的培养介质对组织工程皮肤的影响,HE染色切片观察培养皮肤结构形态。以酮洛芬及其异丙酯为模型药物研究经皮渗透和代谢特性。结果HaCaT细胞经气-液界面培养可形成类表皮层,但不能形成完整的角质层。维生素C可明显促进细胞增殖,维生素D3可促进细胞分化,雌二醇对此组织工程皮肤没有明显的影响。同皮肤细胞匀浆代谢相似,酮洛芬异丙酯被代谢成原药酮洛芬。结论HaCaT细胞在气-液界面培养条件下可形成多层分化不完全的表皮层,保留了一定的酶活性,可用于药物的经皮渗透和代谢等研究。  相似文献   

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目的 研究皮肤角质形成细胞中辣椒素受体TRPV1激活后细胞中蛋白质组学的变化,并鉴定差异蛋白质。方法 皮肤角质形成细胞株HaCaT细胞经辣椒素(CAP)刺激24 h后,提取蛋白样品,采用双向凝胶电泳技术分离HaCaT细胞总蛋白,用PDQuest软件分析对照组和CAP刺激组蛋白组图谱的表达差异点,再运用串联飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白。结果 CAP刺激HaCaT细胞后,磷酸甘油醛异构酶、烯醇化酶、6-磷酸葡萄糖酸内酯酶和组织蛋白酶D蛋白表达水平增高,角蛋白14和60S酸性核糖体蛋白P2表达量降低,这些蛋白可能在TRPV1激活后影响细胞功能的过程中有重要作用。结论 利用蛋白质组学技术,研究皮肤角质形成细胞株HaCaT细胞中TRPV1受体激活后蛋白表达差异,为研究TRPV1受体的作用机制,以及为研发针对性药物,提供新的线索和方向。  相似文献   

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苦参碱对P物质诱导HaCaT细胞表达IFN-γ和IL-10的影响   总被引:3,自引:1,他引:3  
目的:研究苦参碱对P物质诱导表皮角质形成细胞系HaCaT细胞表达IFN-γ和IL-10的影响。方法:采用ELISA法测定P物质诱导HaCaT细胞分泌IFN-γ和IL-10的时效关系与量效关系。以P物质刺激HaCaT细胞,加入不同剂量的苦参碱共孵育24 h,测定苦参碱对IFN-γ和IL-10表达的影响。结果:以1×10-8mol/L P物质刺激HaCaT细胞24 h,可以显著增加HaCaT细胞IFN-γ和IL-10的分泌量,不同浓度的苦参碱均可以促进HaCaT细胞分泌IFN-γ,其中50μg/mL和100μg/mL的苦参碱均可以显著增加IFN-γ的分泌量;但苦参碱对P物质诱导的IL-10的分泌无显著影响。结论:苦参碱可能通过促进抗炎因子IFN-γ的分泌而发挥抗皮肤变态作用。  相似文献   

8.
目的观测表皮细胞抑素对人皮肤角质形成细胞系(HaCaT细胞)和鸡胚表皮细胞的抑制作用。方法应用四甲基偶氮唑蓝微量酶反应比色法(MTT法)观察表皮细胞抑素对HaCaT和鸡胚表皮细胞增殖的抑制作用。结果HaCaT和鸡胚表皮细胞(1×105/ml)培养24h后,分别加入0·25mg/ml、0·125mg/ml、0·0625mg/ml的表皮细胞抑素100μl,48、72h后显示表皮细胞抑素显著的抑制了这两种细胞的生长,并呈量效关系。结论表皮细胞抑素能显著的抑制HaCaT细胞和鸡胚表皮细胞的生长,并呈量效关系。  相似文献   

9.
酮洛芬异丙酯在皮肤细胞中的代谢   总被引:11,自引:0,他引:11  
目的:研究酮洛芬异丙酯在皮肤细胞中的代谢作用,为进一步研究利用酯类前体药物方法改善药物的经皮吸收特性提供实验依据。方法:将人包皮的第3代角质形成细胞或成纤维细胞超声破碎制成匀浆,加入不同量的酮洛芬异丙酯进行37℃温孵实验,通过高效液以谱法分别在不同时间测定细胞匀浆中酮洛芬异丙酯及酮洛芬的浓度。结果:酮洛芬异丙酯在表皮角质形成细胞和真皮成纤维细胞的匀浆中存在代谢现象,有前者的代谢能力明显大于后者。结论:酯类前体药物可在皮肤细胞中被代谢为活性母体药物本身。  相似文献   

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目的通过研究丹皮酚对TNF-α诱导的真皮成纤维细胞MMP-9 mRNA及细胞因子表达的影响,探讨丹皮酚抗皮肤炎症的可能机制。方法采用RT-PCR法考察正常人真皮成纤维细胞中MMP-9 mRNA表达情况以及丹皮酚对TNF-α诱导的真皮成纤维细胞MMP-9 mRNA表达的影响。采用ELISA法测定丹皮酚对TNF-α诱导的真皮成纤维细胞表达IL-1β、IL-6及IL-8的影响。结果MMP-9在正常真皮成纤维细胞上表达极弱,TNF-α可以诱导MMP-9 mRNA的表达,丹皮酚可以抑制TNF-α诱导引起的MMP-9 mRNA表达的上调。成纤维细胞可分泌少量的IL-1β、IL-6及IL-8,TNF-α可以明显增加成纤维细胞IL-1β和IL-8的分泌量,丹皮酚可以抑制TNF-α诱导的IL-1β和IL-8的产生,但对IL-6的分泌无影响。结论丹皮酚可以抑制TNF-α诱导真皮成纤维细胞引起的MMP-9 mRNA及炎症因子IL-1β和IL-8表达水平的上调。  相似文献   

11.
Matrine is a kind of alkaloid found in certain Sophora plants, which has been extensively used in China for the treatment of viral hepatitis, cancer, cardiac diseases and skin diseases (such as atopic dermatitis and eczema). It also has been confirmed that substance P (SP) and its receptor (neurokinin-1 receptor, NK-1R) are involved in the pathogenesis of inflammatory skin disorders. So the present study was designed to investigate the effect of matrine on the expression of NK-1R and cytokines production induced by SP in HaCaT cells (a human epidermal keratinocyte cell line) and dermal fibroblasts. In addition, cell viability was also evaluated. The results showed that matrine inhibited the expression of NK-1R in HaCaT cells and fibroblasts. SP induced the production of interleukin (IL)-1beta, IL-8, interferon (IFN)-gamma, and monocyte chemotactic protein (MCP)-1 in both cell types. Matrine 5-100 microg/mL had little effect on cell viability. It inhibited SP-induced IL-1beta, IL-8 and MCP-1 production in HaCaT cells and fibroblasts, while it increased the production of IFN-gamma in HaCaT cells. Both SP and matrine had no effect on the secretion of IL-6. These findings suggest that matrine may have potential treatment function on SP related cutaneous inflammation by inhibition of the expression of substance P receptor and regulation of the production of inflammatory cytokines.  相似文献   

12.
Endogenous carbohydrates, especially oligo- and polysaccharides, participate in the regulation of a broad range of biological activities, e. g., signal transduction, inflammation, fertilisation, cell-cell-adhesion and act as in vivo markers for the determination of cell types. In the present study, water-soluble (WS) and gel-forming polysaccharides (GF) of ispaghula seed husk (Plantago ovata Forsskal, Plantaginaceae) were characterised as neutral and acidic arabinoxylans and tested under in vitro conditions for regulating activities on cell physiology of human keratinocytes and human primary fibroblasts. Only water-soluble polysaccharides exhibited strong and significant effects on cell physiology of keratinocytes and fibroblasts. Proliferation of cells of the spontaneously immortalised keratinocyte cell line HaCaT was significantly up-regulated in a dose-independent manner. Analysis of activated signal pathways by RNA analysis proved an effect of the acidic arabinoxylan on the expression of keratinocyte growth factor (KGF) in HaCaT cells. Differentiation behaviour of normal human keratinocytes (NHK) determined by involucrin was slightly influenced, due to the enhanced cell proliferation, leading to a cell-cell-mediated indirect induction of early differentiation. WS did not influence late differentiation, as determined by keratin K1 and K10 titres.  相似文献   

13.
Malic acid (MA) has been commonly used in cosmetic products, but the safety reports in skin are sparse. To investigate the biological effects of MA in human skin keratinocytes, we investigated the potential cytotoxicity and apoptotic effects of MA in human keratinocyte cell lines (HaCaT). The data showed that MA induced apoptosis based on the observations of DAPI staining, DNA fragmentation, and sub-G1 phase in HaCaT cells and normal human epidermal keratinocytes (NHEKs). Flow cytometric assays also showed that MA increased the production of mitochondrial superoxide (mito-SOX) but decreased the mitochondrial membrane potential. Analysis of bioenergetics function with the XF 24 analyzer Seahorse extracellular flux analyzer demonstrated that oxygen consumption rate (OCR) was significantly decreased whereas extracellular acidification rate (ECAR) was increased in MA-treated keratinocytes. The occurrence of apoptosis was proved by the increased expressions of FasL, Fas, Bax, Bid, caspases-3, -8, -9, cytochrome c, and the declined expressions of Bcl-2, PARP. MA also induced endoplasmic reticulum stress associated protein expression such as GRP78, GADD153, and ATF6α. We demonstrated that MA had anti-proliferative effect in HaCaT cell through the inhibition of cell cycle progression at G0/G1, and the induction of programmed cell death through endoplasmic reticulum stress- and mitochondria-dependent pathways.  相似文献   

14.
Mutations to the p53 gene are common in UV-exposed keratinocytes and contribute to apoptotic resistance in skin cancer. P53-dependent activity is modulated, in part, by a complex, self-limiting feedback loop imposed by miR-34a-mediated regulation of the lysine deacetylase, SIRT1. Expression of numerous microRNAs is dysregulated in squamous and basal cell carcinomas; however the contribution of specific microRNAs to the pathogenesis of skin cancer remains untested. Through use of RNAi, miRNA target site blocking oligonucleotides and small molecule inhibitors, this study explored the influence of p53 mutational status, SIRT1 activity and miR-34a levels on apoptotic sensitivity in primary (NHEK) and p53-mutated (HaCaT) keratinocyte cell lines. SIRT1 and p53 are overexpressed in p53-mutated keratinocytes, whilst miR-34a levels are 90% less in HaCaT cells. HaCaTs have impaired responses to p53/SIRT1/miR-34a axis manipulation which enhanced survival during exposure to the chemotherapeutic agent, camptothecin. Inhibition of SIRT1 activity in this cell line increased p53 acetylation and doubled camptothecin-induced cell death. Our results demonstrate that p53 mutations increase apoptotic resistance in keratinocytes by interfering with miR-34a-mediated regulation of SIRT1 expression. Thus, SIRT1 inhibitors may have a therapeutic potential for overcoming apoptotic resistance during skin cancer treatment.  相似文献   

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Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging.  相似文献   

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