Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In thi...Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In this study,we investigated the effect of PCF on ultraviolet B(UVB)-induced apoptosis of HaCaT cells and the intracellular signaling pathways involved.Pretreatment with the inducible nitric oxide synthase(iNOS) inhibitor S-methylisothiourea sulfate inhibited UVB-induced apoptosis,indicating that iNOS and NO play important roles in apoptosis.On the other hand,the inhibition of UVB-induced apoptosis in the immortalized keratinocyte(HaCaT) cells by PCF was estimated using a DNA ladder.PCF treatment inhibited UVB-induced iNOS activation,as determined by RT-PCR,NO production,as determined by ESR,and up-regulated heat shock protein(HSP) 90 activation,as determined by Western blotting.Our results indicate that iNOS and NO are involved in UVB-induced apoptosis of HaCaT cells and the protective effect of PCF against UVB irradiation is exerted by suppressing the expression of iNOS,followed by inhibition of NO release and enhanced activation of HSP90.展开更多
MicroRNAs （miRNAs） are 21 to 24 nucleotide, non-coding RNA molecules that post-transcriptionally regulate the expression of target genes. Ultraviolet B （UVB） radiation has been shown to inhibit phosphatase and ten...MicroRNAs （miRNAs） are 21 to 24 nucleotide, non-coding RNA molecules that post-transcriptionally regulate the expression of target genes. Ultraviolet B （UVB） radiation has been shown to inhibit phosphatase and tensin homolog deleted on chromosome 10 （PTEN） expression in HaCaT cells through an unknown mechanism. In this study, we investigated whether miR-141 can regulate UVB exposure-mediated inhibition of PTEN expression. Real-time RT-PCR, annexin V/fluorescein isothiocyanate staining, Western blotting and anti-miRNA oligonucleotide transfection were employed in this study. We found that upregulation of miR-141 expression after UVB irradiation was inversely correlated with PTEN expression levels in HaCaT cells. Furthermore, miR-141 expression increased apoptosis, while anti-miR-141 partly restored PTEN expression and reversed the pro-apoptosis effect of UVB. UVB suppresses the expression of PTEN by upregulating miR-141 in HaCaT cells. Therefore, miR-141 is a potential gene therapy target for UVB-induced photodamage.展开更多
Objective： To study the effect of oxymatrine on the expression of nuclear factor-kappa B（NF-K B） of Human benign epidermal keratinocytes line（HaCaT cells）. Methods：HaCaT cells were cultured with different concen...Objective： To study the effect of oxymatrine on the expression of nuclear factor-kappa B（NF-K B） of Human benign epidermal keratinocytes line（HaCaT cells）. Methods：HaCaT cells were cultured with different concentration of Oxymatrine（10 μ g/ml, 50 μ g/ ml and 100 μg/ml） for 24 h, then 10.5 mol/L Substance P was added to the cells. After 30 min, NF-K B expression in the cells was observed by immunocytochemistry, NF-K B P65 protein expression was evaluated by Western blot, and the mRNA expression of NF-K B P65 was evaluated by reverse transcription polymerase chain reaction（RT-PCR）. The 10.5 mol/L Substance P and culture medium were added to the Substance P group and control group, respectively. Results：In control group, expression rate of positive cells, the expressions of protein and mRNA of NF-K B were all low. In Substance P group, when 10μ mol/L Substance P was added, the expressions were all increased（P 〈 0.05）. But in Oxymatrine groups, the expression rate of positive cells, the expressions of protein and mRNA were all descended in a concentration-dependent manner（P 〈 0.05 or P 〈 0.01）. Conclusion：Oxymatrine can down-regulate the expression of NF-K B of the HaCaT cells and may play an important role in regulating anti-inflammation and immunity.展开更多
Objective Ultraviolet B(UVB)mainly acts on the skin epidermis,causing oxidative damage and apoptosis of keratinocytes.Jin Bai Mei Yan Prescription(JBMYP)comprises a variety of antioxidant traditional Chinese medicines...Objective Ultraviolet B(UVB)mainly acts on the skin epidermis,causing oxidative damage and apoptosis of keratinocytes.Jin Bai Mei Yan Prescription(JBMYP)comprises a variety of antioxidant traditional Chinese medicines(TCM).In this study,we aimed to evaluate the effects of JBMYP on the oxidative damage induced by UVB in human immortalized epidermal keratinocytes(HaCaT)cells.Methods HaCaT cells were divided into six groups:control group,model(UVB)group,positive(UVB+vitamin E)group,UVB+JBMYP low dose group(160μg/mL),UVB+JBMYP moderate dose group(800μg/mL),and UVB+JBMYP high dose group(1600μg/mL).HaCaT cells were irradiated with UVB and treated with JBMYP for 24 h.Methyl thiazolyl tetrazolium(MTT)assay and real-time unlabeled cell function analyzer were used to assess the cell survival and proliferation rates,respectively.At the same time,the levels of intracellular reactive oxygen species(ROS),lipid peroxide malondialdehyde(MDA),glutathione(GSH),and hydroxyproline(HYP),as well as the activities of antioxidant enzyme superoxide dismutase(SOD)and catalase(CAT)were evaluated using enzyme linked immunosorbent assay(ELISA).Results Compared with the model group,the survival rate of HaCaT cells in each dosage group of JBMYP was significantly improved(P<0.05).Further,JBMYP could promote the proliferation of HaCaT cells,leading to a reduction in the contents of MDA and ROS,and increase in the contents of SOD,CAT,GSH and HYP in HaCaT cells.Conclusions JBMYP has enhanced protective effect on oxidative damage induced by UVB in HaCaT cells.展开更多
Objective： To observe the effects of different doses of acitretin on the transcription of FGF10 mRNA and the translation of FGF10 protein in cultured HaCaT cells. Methods： HaCaT cells were treated with different dos...Objective： To observe the effects of different doses of acitretin on the transcription of FGF10 mRNA and the translation of FGF10 protein in cultured HaCaT cells. Methods： HaCaT cells were treated with different doses of acitretin for 48 h, then the changes on the transcription of FGF10 mRNA and the translation of FGF10 protein in these cells were detected by immunofluorescence and in situ hybridization assay. Results： Compared with the control group, the transcription of FGF10 mRNA and the translation of FGF10 protein were gradually decreased along with the increasing dose of aeitretin. There were significant differences between different groups （P〈0.01）. Conclusion： Aeitretin could inhibit the transcription of FGF10 mRNA and the translation of FGF10 protein in HaCaT cells. With the dose of aeitretin increased, the stains of both FGF10 mRNA and FGF10 protein in HaCaT cells are reduced.展开更多
Objective Vitamin D and Toll-like receptor-4(TLR-4)inhibition are involved in the protection of keratinocytes.The effects of combination of 1,25(OH)_(2)D_(3) and TLR-4 inhibitor on the protection of keratinocytes agai...Objective Vitamin D and Toll-like receptor-4(TLR-4)inhibition are involved in the protection of keratinocytes.The effects of combination of 1,25(OH)_(2)D_(3) and TLR-4 inhibitor on the protection of keratinocytes against ultraviolet radiation B(UVB)irradiation remain unclear.This study was undertaken to explore the effects of combination of 1,25(OH)_(2)D_(3) and TAK-242(TLR-4 inhibitor)on the damage to HaCaT cells caused by UVB irradiation.Methods In vitro,HaCaT cells were treated with 1,25(OH)_(2)D_(3) or/and TAK-242 prior to UVB irradiation at the intensity of 20 mJ/cm^(2),then the production of reactive oxygen species(ROS),cell migration,apoptosis of cells,and the expression of oxidative stress,endoplasmic reticulum stress,and apoptosis related proteins were determined.Results Compared with the HaCaT cells treated with 1,25(OH)_(2)D_(3) or TAK-242,the cells treated with both 1,25(OH)_(2)D_(3) and TAK-242 showed,1)significantly lower production of ROS(P<0.05);2)significantly less apoptosis of HaCaT cells(P<0.05);3)significantly lower expression of NF-κB,Caspase-8,Cyto-C,Caspase-3(P<0.05).Conclusion The combination of 1,25(OH)_(2)D_(3) and TAK-242 could produce a better protection for HaCaT cells via inhibiting the oxidative stress,endoplasmic reticulum stress and apoptosis than 1,25(OH)_(2)D_(3) or TAK-242 alone.展开更多
<strong>Background:</strong> Urban air pollution contributes to lung and cardiovascular system dysfunction, making it a major concern for human health. Its impact on skin integrity, associated with increas...<strong>Background:</strong> Urban air pollution contributes to lung and cardiovascular system dysfunction, making it a major concern for human health. Its impact on skin integrity, associated with increased occurrence of atopic dermatitis, is now recognized, but its cellular mechanisms remain poorly understood. <strong>Objective:</strong> In the present study we aimed at establishing the impact of urban pollutant on mitochondrial dynamics and bioenergetics using the HaCaT cell model. We also sought to establish the protective effect of ECH-5195 (red <em>Panax ginseng</em> extract), standardized in ginsenosides, in reversing pollution-induced mitochondrial defects. <strong>Methods:</strong> Urban pollution exposure was mimicked by 1 h exposure of HaCaT cells with standardized atmospheric particulate matter containing PAHs, nitro-PAHs, PCB congeners, and chlorinated pesticides with a mean particulate diameter of 5.85 μm (SRM1648). <strong>Results:</strong> The presence of urban pollutant in the cultures increased the prevalence of hyperfission by 1.41-fold (p = 0.023) and fission by 1.35 fold (p = 0.006) in the reticular mitochondrial network. ECH-5195 reduced both pollution-induced hyperfission by 0.54-fold (p = 0.004) and fission by 0.68-fold (p = 0.0006) normalizing the mitochondrial reticular network. Pollution exposure was associated with a significant reduction of basal OCR and increased lactate production, pushing the cell to rely on glycolysis for ATP production. When ECH-5195 was used, OCR was significantly increased, and the glycolytic contribution to ATP production was reduced while both oxidative phosphorylation and mitochondrial respiration were increased demonstrating mitochondrial re-engagement in ATP production. <strong>Conclusions:</strong> Pollution exposure was disruptive for both the mitochondrial network dynamics and mitochondrial respiration. Ginsenosides in ECH-5195 efficiently protected both from pollution-induced defects.展开更多
Psoriasis is a chronic,refractory,inflammatory skin disease that occurs in young adults.The traditional animal model cannot simulate the skin characteristics of patients with psoriasis effectively,so it is difficult t...Psoriasis is a chronic,refractory,inflammatory skin disease that occurs in young adults.The traditional animal model cannot simulate the skin characteristics of patients with psoriasis effectively,so it is difficult to be used for in-depth study of psoriasis mechanism.Immortalized human epidermal cells(HaCaT) is a non-tumor,immortalized human epidermal cell which is widely used in the study of dermatosis.HaCaT cells are the best choice for the study of psoriasis mechanism because their immu.nological characteristics and reproductive ability are coincide with the pathological features of psoriasis.This article reviews the specific methods such as establishment of cell method,cytokine and chemo.kine analysisin the pathogenesis study of psoriasis based on HaCaT cells,hoping to provide some thoughts for drug′s pharmacological activity research.展开更多
Objective:To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (HaCat cell),particularly the effect of mitomycin on intracellular messenger RNA (mRNA) ...Objective:To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (HaCat cell),particularly the effect of mitomycin on intracellular messenger RNA (mRNA) synthesis of collagen and growth factors of fibroblast.Methods:The normal dermal fibroblast and HaCat cell were cultured in vitro.Cell cultures were exposed to 0.4 and 0.04 mg/ml of mitomycin solution,and serum-free culture medium was used as control.The cellular morphology change,growth characteristics,cell proliferation,and apoptosis were observed at different intervals.For the fibroblasts,the mRNA expression changes of transforming growth factor (TGF)-β1,basic fibroblast growth factor (bFGF),procollagen Ⅰ,and Ⅲ were detected by reverse transcription polymerase chain reaction (RT-PCR).Results:The cultured normal human skin fibroblast and HaCat cell grew exponentially.A 5-min exposure to mitomycin at either 0.4 or 0.04 mg/ml caused marked dose-dependent cell proliferation inhibition on both fibroblasts and HaCat cells.Cell morphology changed,cell density decreased,and the growth curves were without an exponential phase.The fibroblast proliferated on the 5th day after the 5-min exposure of mitomycin at 0.04 mg/ml.Meanwhile,5-min application of mitomycin at either 0.04 or 0.4 mg/ml induced fibroblast apoptosis but not necrosis.The apoptosis rate of the fibroblast increased with a higher concentration of mytomycin (p<0.05).A 5-min exposure to mitomycin at 0.4 mg/ml resulted in a marked decrease in the mRNA production of TGF-β1,procollagen Ⅰ and Ⅲ,and a marked increase in the mRNA production of bFGF.Conclusions:Mitomycin can inhibit fibroblast proliferation,induce fibroblast apoptosis,and regulate intracellular protein expression on mRNA levels.In additon,mitomycin can inhibit HaCat cell proliferation,so epithelial cell needs more protecting to avoid mitomycin's side effect when it is applied clinically.展开更多
The bioeffects of silica nanoparticles (SiNP), phosphorylate-terminated nanoparticles (PO4- NP) and amino-terminated nanoparticles (NH2NP) on HaCaT cell line have been studied in this paper. The effects of the three k...The bioeffects of silica nanoparticles (SiNP), phosphorylate-terminated nanoparticles (PO4- NP) and amino-terminated nanoparticles (NH2NP) on HaCaT cell line have been studied in this paper. The effects of the three kinds of functionalized silica nanoparticles on adherence, proliferation and cycle of HaCaT cells have been investigated. And the cel- lular uptake of the three kinds of functionalized silica nanoparticles by HaCaT cells has also been exam- ined. Results indicated that the bioeffects of the three kinds of functionalized nanoparticles on HaCaT cells were concentration-dependent. And the three kinds of functionalized nanoparticles all exhibited well bio- compatibility if the concentration was below 0.2 μg/μL. While the cytotoxicities of the three kinds of function- alized nanoparticles on HaCaT cells would increase with the increasing of nanoparticles concentration, and the following order was observed: NH2NP > SiNP > PO4NP. In addition, the quantity and rapidity of cellular uptake of nanoparticles by HaCaT cells were diverse due to the different functional groups. Under the same conditions, NH2NP was most and fast internalized by HaCaT cells, followed by SiNP, and PO4NP was the least and slowest. These results provided theoretical foundation for the safe applica- tion and further modification of silica nanoparticles, which could broaden the application of silica nano- particles in biomedicine.展开更多
基金Supported by the National Natural Science Foundation of China (30471458)National Natural Science Foundation of Shandong Province (Z2007c09)
文摘Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In this study,we investigated the effect of PCF on ultraviolet B(UVB)-induced apoptosis of HaCaT cells and the intracellular signaling pathways involved.Pretreatment with the inducible nitric oxide synthase(iNOS) inhibitor S-methylisothiourea sulfate inhibited UVB-induced apoptosis,indicating that iNOS and NO play important roles in apoptosis.On the other hand,the inhibition of UVB-induced apoptosis in the immortalized keratinocyte(HaCaT) cells by PCF was estimated using a DNA ladder.PCF treatment inhibited UVB-induced iNOS activation,as determined by RT-PCR,NO production,as determined by ESR,and up-regulated heat shock protein(HSP) 90 activation,as determined by Western blotting.Our results indicate that iNOS and NO are involved in UVB-induced apoptosis of HaCaT cells and the protective effect of PCF against UVB irradiation is exerted by suppressing the expression of iNOS,followed by inhibition of NO release and enhanced activation of HSP90.
基金supported by the National Natural Science Foundationof China (No. 30771946 and No. 81000700)
文摘MicroRNAs （miRNAs） are 21 to 24 nucleotide, non-coding RNA molecules that post-transcriptionally regulate the expression of target genes. Ultraviolet B （UVB） radiation has been shown to inhibit phosphatase and tensin homolog deleted on chromosome 10 （PTEN） expression in HaCaT cells through an unknown mechanism. In this study, we investigated whether miR-141 can regulate UVB exposure-mediated inhibition of PTEN expression. Real-time RT-PCR, annexin V/fluorescein isothiocyanate staining, Western blotting and anti-miRNA oligonucleotide transfection were employed in this study. We found that upregulation of miR-141 expression after UVB irradiation was inversely correlated with PTEN expression levels in HaCaT cells. Furthermore, miR-141 expression increased apoptosis, while anti-miR-141 partly restored PTEN expression and reversed the pro-apoptosis effect of UVB. UVB suppresses the expression of PTEN by upregulating miR-141 in HaCaT cells. Therefore, miR-141 is a potential gene therapy target for UVB-induced photodamage.
文摘Objective： To study the effect of oxymatrine on the expression of nuclear factor-kappa B（NF-K B） of Human benign epidermal keratinocytes line（HaCaT cells）. Methods：HaCaT cells were cultured with different concentration of Oxymatrine（10 μ g/ml, 50 μ g/ ml and 100 μg/ml） for 24 h, then 10.5 mol/L Substance P was added to the cells. After 30 min, NF-K B expression in the cells was observed by immunocytochemistry, NF-K B P65 protein expression was evaluated by Western blot, and the mRNA expression of NF-K B P65 was evaluated by reverse transcription polymerase chain reaction（RT-PCR）. The 10.5 mol/L Substance P and culture medium were added to the Substance P group and control group, respectively. Results：In control group, expression rate of positive cells, the expressions of protein and mRNA of NF-K B were all low. In Substance P group, when 10μ mol/L Substance P was added, the expressions were all increased（P 〈 0.05）. But in Oxymatrine groups, the expression rate of positive cells, the expressions of protein and mRNA were all descended in a concentration-dependent manner（P 〈 0.05 or P 〈 0.01）. Conclusion：Oxymatrine can down-regulate the expression of NF-K B of the HaCaT cells and may play an important role in regulating anti-inflammation and immunity.
基金funding support from the National Administration of Traditional Chinese Medicine(No.2017-149-11)National Base for International Cooperation(No.2016-65)Henan Province Industry-University-Research Collaboration(No.182107000029)to conduct the Special Project on Standardization of Traditional Chinese Medicine。
文摘Objective Ultraviolet B(UVB)mainly acts on the skin epidermis,causing oxidative damage and apoptosis of keratinocytes.Jin Bai Mei Yan Prescription(JBMYP)comprises a variety of antioxidant traditional Chinese medicines(TCM).In this study,we aimed to evaluate the effects of JBMYP on the oxidative damage induced by UVB in human immortalized epidermal keratinocytes(HaCaT)cells.Methods HaCaT cells were divided into six groups:control group,model(UVB)group,positive(UVB+vitamin E)group,UVB+JBMYP low dose group(160μg/mL),UVB+JBMYP moderate dose group(800μg/mL),and UVB+JBMYP high dose group(1600μg/mL).HaCaT cells were irradiated with UVB and treated with JBMYP for 24 h.Methyl thiazolyl tetrazolium(MTT)assay and real-time unlabeled cell function analyzer were used to assess the cell survival and proliferation rates,respectively.At the same time,the levels of intracellular reactive oxygen species(ROS),lipid peroxide malondialdehyde(MDA),glutathione(GSH),and hydroxyproline(HYP),as well as the activities of antioxidant enzyme superoxide dismutase(SOD)and catalase(CAT)were evaluated using enzyme linked immunosorbent assay(ELISA).Results Compared with the model group,the survival rate of HaCaT cells in each dosage group of JBMYP was significantly improved(P<0.05).Further,JBMYP could promote the proliferation of HaCaT cells,leading to a reduction in the contents of MDA and ROS,and increase in the contents of SOD,CAT,GSH and HYP in HaCaT cells.Conclusions JBMYP has enhanced protective effect on oxidative damage induced by UVB in HaCaT cells.
基金Supported by the Shaanxi Youth Science Fund (No.2003K10-G56)
文摘Objective： To observe the effects of different doses of acitretin on the transcription of FGF10 mRNA and the translation of FGF10 protein in cultured HaCaT cells. Methods： HaCaT cells were treated with different doses of acitretin for 48 h, then the changes on the transcription of FGF10 mRNA and the translation of FGF10 protein in these cells were detected by immunofluorescence and in situ hybridization assay. Results： Compared with the control group, the transcription of FGF10 mRNA and the translation of FGF10 protein were gradually decreased along with the increasing dose of aeitretin. There were significant differences between different groups （P〈0.01）. Conclusion： Aeitretin could inhibit the transcription of FGF10 mRNA and the translation of FGF10 protein in HaCaT cells. With the dose of aeitretin increased, the stains of both FGF10 mRNA and FGF10 protein in HaCaT cells are reduced.
基金supported by the Department of Science and Technology of Jilin Province.[Grant No:20210204024YY]。
文摘Objective Vitamin D and Toll-like receptor-4(TLR-4)inhibition are involved in the protection of keratinocytes.The effects of combination of 1,25(OH)_(2)D_(3) and TLR-4 inhibitor on the protection of keratinocytes against ultraviolet radiation B(UVB)irradiation remain unclear.This study was undertaken to explore the effects of combination of 1,25(OH)_(2)D_(3) and TAK-242(TLR-4 inhibitor)on the damage to HaCaT cells caused by UVB irradiation.Methods In vitro,HaCaT cells were treated with 1,25(OH)_(2)D_(3) or/and TAK-242 prior to UVB irradiation at the intensity of 20 mJ/cm^(2),then the production of reactive oxygen species(ROS),cell migration,apoptosis of cells,and the expression of oxidative stress,endoplasmic reticulum stress,and apoptosis related proteins were determined.Results Compared with the HaCaT cells treated with 1,25(OH)_(2)D_(3) or TAK-242,the cells treated with both 1,25(OH)_(2)D_(3) and TAK-242 showed,1)significantly lower production of ROS(P<0.05);2)significantly less apoptosis of HaCaT cells(P<0.05);3)significantly lower expression of NF-κB,Caspase-8,Cyto-C,Caspase-3(P<0.05).Conclusion The combination of 1,25(OH)_(2)D_(3) and TAK-242 could produce a better protection for HaCaT cells via inhibiting the oxidative stress,endoplasmic reticulum stress and apoptosis than 1,25(OH)_(2)D_(3) or TAK-242 alone.
文摘<strong>Background:</strong> Urban air pollution contributes to lung and cardiovascular system dysfunction, making it a major concern for human health. Its impact on skin integrity, associated with increased occurrence of atopic dermatitis, is now recognized, but its cellular mechanisms remain poorly understood. <strong>Objective:</strong> In the present study we aimed at establishing the impact of urban pollutant on mitochondrial dynamics and bioenergetics using the HaCaT cell model. We also sought to establish the protective effect of ECH-5195 (red <em>Panax ginseng</em> extract), standardized in ginsenosides, in reversing pollution-induced mitochondrial defects. <strong>Methods:</strong> Urban pollution exposure was mimicked by 1 h exposure of HaCaT cells with standardized atmospheric particulate matter containing PAHs, nitro-PAHs, PCB congeners, and chlorinated pesticides with a mean particulate diameter of 5.85 μm (SRM1648). <strong>Results:</strong> The presence of urban pollutant in the cultures increased the prevalence of hyperfission by 1.41-fold (p = 0.023) and fission by 1.35 fold (p = 0.006) in the reticular mitochondrial network. ECH-5195 reduced both pollution-induced hyperfission by 0.54-fold (p = 0.004) and fission by 0.68-fold (p = 0.0006) normalizing the mitochondrial reticular network. Pollution exposure was associated with a significant reduction of basal OCR and increased lactate production, pushing the cell to rely on glycolysis for ATP production. When ECH-5195 was used, OCR was significantly increased, and the glycolytic contribution to ATP production was reduced while both oxidative phosphorylation and mitochondrial respiration were increased demonstrating mitochondrial re-engagement in ATP production. <strong>Conclusions:</strong> Pollution exposure was disruptive for both the mitochondrial network dynamics and mitochondrial respiration. Ginsenosides in ECH-5195 efficiently protected both from pollution-induced defects.
文摘Psoriasis is a chronic,refractory,inflammatory skin disease that occurs in young adults.The traditional animal model cannot simulate the skin characteristics of patients with psoriasis effectively,so it is difficult to be used for in-depth study of psoriasis mechanism.Immortalized human epidermal cells(HaCaT) is a non-tumor,immortalized human epidermal cell which is widely used in the study of dermatosis.HaCaT cells are the best choice for the study of psoriasis mechanism because their immu.nological characteristics and reproductive ability are coincide with the pathological features of psoriasis.This article reviews the specific methods such as establishment of cell method,cytokine and chemo.kine analysisin the pathogenesis study of psoriasis based on HaCaT cells,hoping to provide some thoughts for drug′s pharmacological activity research.
文摘Objective:To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (HaCat cell),particularly the effect of mitomycin on intracellular messenger RNA (mRNA) synthesis of collagen and growth factors of fibroblast.Methods:The normal dermal fibroblast and HaCat cell were cultured in vitro.Cell cultures were exposed to 0.4 and 0.04 mg/ml of mitomycin solution,and serum-free culture medium was used as control.The cellular morphology change,growth characteristics,cell proliferation,and apoptosis were observed at different intervals.For the fibroblasts,the mRNA expression changes of transforming growth factor (TGF)-β1,basic fibroblast growth factor (bFGF),procollagen Ⅰ,and Ⅲ were detected by reverse transcription polymerase chain reaction (RT-PCR).Results:The cultured normal human skin fibroblast and HaCat cell grew exponentially.A 5-min exposure to mitomycin at either 0.4 or 0.04 mg/ml caused marked dose-dependent cell proliferation inhibition on both fibroblasts and HaCat cells.Cell morphology changed,cell density decreased,and the growth curves were without an exponential phase.The fibroblast proliferated on the 5th day after the 5-min exposure of mitomycin at 0.04 mg/ml.Meanwhile,5-min application of mitomycin at either 0.04 or 0.4 mg/ml induced fibroblast apoptosis but not necrosis.The apoptosis rate of the fibroblast increased with a higher concentration of mytomycin (p<0.05).A 5-min exposure to mitomycin at 0.4 mg/ml resulted in a marked decrease in the mRNA production of TGF-β1,procollagen Ⅰ and Ⅲ,and a marked increase in the mRNA production of bFGF.Conclusions:Mitomycin can inhibit fibroblast proliferation,induce fibroblast apoptosis,and regulate intracellular protein expression on mRNA levels.In additon,mitomycin can inhibit HaCat cell proliferation,so epithelial cell needs more protecting to avoid mitomycin's side effect when it is applied clinically.
基金This work was supported by the Key Project of Natural Science Foundation of China （Grant No. 20135010）National Key Basic Research Program （Grant No. 2002CB513100-10）+2 种基金Key Technologies Research and Development Program （Grant No. 2003 BA310A16）High-Tech Research and Development （863） Program （Grant No. 2003AA302250） the National Natural Science Foundation of China （Grant No. 20405005）.
文摘The bioeffects of silica nanoparticles (SiNP), phosphorylate-terminated nanoparticles (PO4- NP) and amino-terminated nanoparticles (NH2NP) on HaCaT cell line have been studied in this paper. The effects of the three kinds of functionalized silica nanoparticles on adherence, proliferation and cycle of HaCaT cells have been investigated. And the cel- lular uptake of the three kinds of functionalized silica nanoparticles by HaCaT cells has also been exam- ined. Results indicated that the bioeffects of the three kinds of functionalized nanoparticles on HaCaT cells were concentration-dependent. And the three kinds of functionalized nanoparticles all exhibited well bio- compatibility if the concentration was below 0.2 μg/μL. While the cytotoxicities of the three kinds of function- alized nanoparticles on HaCaT cells would increase with the increasing of nanoparticles concentration, and the following order was observed: NH2NP > SiNP > PO4NP. In addition, the quantity and rapidity of cellular uptake of nanoparticles by HaCaT cells were diverse due to the different functional groups. Under the same conditions, NH2NP was most and fast internalized by HaCaT cells, followed by SiNP, and PO4NP was the least and slowest. These results provided theoretical foundation for the safe applica- tion and further modification of silica nanoparticles, which could broaden the application of silica nano- particles in biomedicine.
