利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的...利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。展开更多
目的:研究X射线和紫外线B(UVB)对表皮细胞增殖的影响,以及对褪黑素受体(MTNR)蛋白表达的影响,为生活在高海拔地区可能受到较高电离和非电离辐射的公众辐射效应及其防护提供指导。方法:经照射剂量为0 Gy(对照组)、0.5 Gy、2 Gy和5 Gy(照...目的:研究X射线和紫外线B(UVB)对表皮细胞增殖的影响,以及对褪黑素受体(MTNR)蛋白表达的影响,为生活在高海拔地区可能受到较高电离和非电离辐射的公众辐射效应及其防护提供指导。方法:经照射剂量为0 Gy(对照组)、0.5 Gy、2 Gy和5 Gy(照射组)X射线以及0 m J/cm2、20 m J/cm2、50 m J/cm2、100 m J/cm2和200 m J/cm2的UVB分别照射人表皮细胞Hacat和人黑色素瘤细胞A875,通过四甲基偶氮唑(MTT)和克隆形成率实验检测细胞增殖能力变化,采用蛋白质印迹法检测褪黑素受体MTNR-1B蛋白表达变化。结果:在照射剂量为0.5 Gy、2 Gy和5 Gy的X射线以及20 m J/cm2和50 m J/cm2UVB的照射下,可导致Hacat和A875细胞生长不同程度的降低。3种不同剂量X射线照射后Hacat细胞克隆形成率分别为对照组的71.2%、36.7%和16.3%,与对照组比较差异有统计学意义(F=249.96,P<0.05),A875细胞细胞克隆形成率分别为对照组的64.7%、49.5%和31.6%,与对照组比较差异有统计学意义(F=147.29,P<0.05);在照射剂量为20 m J/cm2和50 m J/cm2的UVB照射后Hacat细胞的克隆形成率分别为对照组的40.74%和12.96%,A875细胞的克隆形成率分别为对照组的16.33%和6.12%;50 m J/cm、100 m J/cm和200 m J/cm2的UVB照射后A875细胞中MTNR1B表达随着剂量的增大而显著增加,而3种不同剂量X射线对MTNR1B表达在不同剂量之间无显著影响。结论:X射线和UVB照射后均可引起细胞增殖和存活能力的降低,UVB照射后引起的细胞增殖抑制可能与褪黑素受体表达变化相关。展开更多
研究蔓越莓对中波紫外线(UVB)诱导的人表皮角质形成细胞(Ha Ca T)光损伤的保护作用。用3个不同剂量的蔓越莓果汁预处理Ha Ca T细胞6 h,采用60 m J/cm^2强度UVB照射细胞;然后用MTT法检测细胞的生存率,吸取细胞上清液采用比色法检测丙二醛...研究蔓越莓对中波紫外线(UVB)诱导的人表皮角质形成细胞(Ha Ca T)光损伤的保护作用。用3个不同剂量的蔓越莓果汁预处理Ha Ca T细胞6 h,采用60 m J/cm^2强度UVB照射细胞;然后用MTT法检测细胞的生存率,吸取细胞上清液采用比色法检测丙二醛(MDA)、羟脯氨酸(Hyp)的含量和超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、乳酸脱氢酶(LDH)的活性变化,用荧光法检测细胞内活性氧自由基(ROS)含量,用倒置显微镜和流式细胞仪观察检测细胞凋亡状况。UVB辐射对Ha Ca T细胞造成比较严重的损伤;相比于空白对照组,UVB辐射模型组Ha Ca T细胞活性下降了29.54%,SOD和GSH-Px活性极显著降低(P<0.01);相比于模型组,蔓越莓各剂量组可显著提高UVB照射后Ha Ca T细胞的活力(P<0.05或P<0.01),提高SOD和GSH-Px活性(P<0.01),减少MDA和ROS含量(P<0.05或P<0.01),并且降低LDH活性和增加Hyp含量(P<0.01),呈剂量依赖关系,从而降低UVB所导致的细胞损伤及凋亡率升高。蔓越莓可以明显减少UVB诱导Ha Ca T细胞的氧化损伤和凋亡,具有显著的光保护功效,其作用机制与增强细胞抗氧化能力、加速清除氧自由基以及促进胶原蛋白合成有关。展开更多
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
The atmosphere protects humans,plants,animals,and microorganisms from damaging doses of ultraviolet-B(UVB) solar radiation(280-320 nm) because it modifies the UVB reaching the Earth's surface.This modification is ...The atmosphere protects humans,plants,animals,and microorganisms from damaging doses of ultraviolet-B(UVB) solar radiation(280-320 nm) because it modifies the UVB reaching the Earth's surface.This modification is a function of the solar radiation's path length through the atmosphere and the amount of each attenuator along the path length.The path length is determined by solar zenith angle(SZA).The present work explains the dependence of hemispherical transmittance of UVB on SZA.The database used consists of five years of hourly UVB and global solar radiation measurements.From 2001 to 2005,the South Valley University(SVU) meteorological research station(26.20°N,32.75°E) carried out these measurements on a horizontal surface.In addition,the corresponding extraterrestrial UVB(UVBext) and broadband solar radiation(Gext) were estimated.Consequently,the hemispherical transmittance of UVB(KtUVB) and the hemispherical transmittance of global solar radiation(Kt) were estimated.Furthermore,the UVB redaction due to the atmosphere was evaluated.An analysis of the dependence between KtUVB and SZA at different ranges of Kt was performed.A functional dependence between KtUVB and SZA(KtUVB=-a(SZA)+b) for very narrow Kt-ranges(width of ranges was 0.01) was developed.The results are discussed,and the sensitivity of △KtUVB to △SZA for very narrow Kt-ranges was studied.It was found that the sensitivity of △KtUVB to △SZA slightly increases with increased Kt,which means KtUVB is sensitive to SZA as Kt increases.The maximum correlation(R) between KtUVB and SZA was equal to-0.83 for Kt= 0.76.展开更多
目的探讨窄谱中波紫外线(NB-UVB)家庭光疗与院内光疗治疗小腿斑块状银屑病的疗效比较。方法选择2018年7月至2020年6月嘉兴市第一医院皮肤科门诊收治的58例小腿斑块状银屑病患者,随机分为对照组28例和观察组30例收治的对照组给予外用卡...目的探讨窄谱中波紫外线(NB-UVB)家庭光疗与院内光疗治疗小腿斑块状银屑病的疗效比较。方法选择2018年7月至2020年6月嘉兴市第一医院皮肤科门诊收治的58例小腿斑块状银屑病患者,随机分为对照组28例和观察组30例收治的对照组给予外用卡泊三醇倍他米松软膏联合院内照射NB-UVB,观察组给予外用卡泊三醇倍他米松软膏联合家中自行照射NB-UVB,两组疗程均为8周。通过治疗前后银屑病皮损面积与严重程度指数(psoriasis area and severity index,PASI)评分判定疗效,记录皮肤不良反应发生情况,以调查问卷形式评价便利性。结果治疗后两组的PASI评分均较治疗前下降,差异有统计学意义(P<0.05);治疗后两组的PASI评分比较,差异无统计学意义(P>0.05)。对照组的总有效率为82.14%,观察组为76.67%,两组比较,差异无统计学意义(P>0.05)。对照组的轻度皮肤不良反应发生率为17.86%,观察组为13.33%,两组比较,差异无统计学意义(P>0.05)。观察组的便利性优于对照组,差异有统计学意义(P<0.05)。结论NB-UVB家庭光疗对比院内光疗治疗小腿斑块状银屑病,疗效相当,安全性良好,便利程度高,值得临床推广。展开更多
Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with human...Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.展开更多
文摘利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。
文摘目的:研究X射线和紫外线B(UVB)对表皮细胞增殖的影响,以及对褪黑素受体(MTNR)蛋白表达的影响,为生活在高海拔地区可能受到较高电离和非电离辐射的公众辐射效应及其防护提供指导。方法:经照射剂量为0 Gy(对照组)、0.5 Gy、2 Gy和5 Gy(照射组)X射线以及0 m J/cm2、20 m J/cm2、50 m J/cm2、100 m J/cm2和200 m J/cm2的UVB分别照射人表皮细胞Hacat和人黑色素瘤细胞A875,通过四甲基偶氮唑(MTT)和克隆形成率实验检测细胞增殖能力变化,采用蛋白质印迹法检测褪黑素受体MTNR-1B蛋白表达变化。结果:在照射剂量为0.5 Gy、2 Gy和5 Gy的X射线以及20 m J/cm2和50 m J/cm2UVB的照射下,可导致Hacat和A875细胞生长不同程度的降低。3种不同剂量X射线照射后Hacat细胞克隆形成率分别为对照组的71.2%、36.7%和16.3%,与对照组比较差异有统计学意义(F=249.96,P<0.05),A875细胞细胞克隆形成率分别为对照组的64.7%、49.5%和31.6%,与对照组比较差异有统计学意义(F=147.29,P<0.05);在照射剂量为20 m J/cm2和50 m J/cm2的UVB照射后Hacat细胞的克隆形成率分别为对照组的40.74%和12.96%,A875细胞的克隆形成率分别为对照组的16.33%和6.12%;50 m J/cm、100 m J/cm和200 m J/cm2的UVB照射后A875细胞中MTNR1B表达随着剂量的增大而显著增加,而3种不同剂量X射线对MTNR1B表达在不同剂量之间无显著影响。结论:X射线和UVB照射后均可引起细胞增殖和存活能力的降低,UVB照射后引起的细胞增殖抑制可能与褪黑素受体表达变化相关。
文摘研究蔓越莓对中波紫外线(UVB)诱导的人表皮角质形成细胞(Ha Ca T)光损伤的保护作用。用3个不同剂量的蔓越莓果汁预处理Ha Ca T细胞6 h,采用60 m J/cm^2强度UVB照射细胞;然后用MTT法检测细胞的生存率,吸取细胞上清液采用比色法检测丙二醛(MDA)、羟脯氨酸(Hyp)的含量和超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、乳酸脱氢酶(LDH)的活性变化,用荧光法检测细胞内活性氧自由基(ROS)含量,用倒置显微镜和流式细胞仪观察检测细胞凋亡状况。UVB辐射对Ha Ca T细胞造成比较严重的损伤;相比于空白对照组,UVB辐射模型组Ha Ca T细胞活性下降了29.54%,SOD和GSH-Px活性极显著降低(P<0.01);相比于模型组,蔓越莓各剂量组可显著提高UVB照射后Ha Ca T细胞的活力(P<0.05或P<0.01),提高SOD和GSH-Px活性(P<0.01),减少MDA和ROS含量(P<0.05或P<0.01),并且降低LDH活性和增加Hyp含量(P<0.01),呈剂量依赖关系,从而降低UVB所导致的细胞损伤及凋亡率升高。蔓越莓可以明显减少UVB诱导Ha Ca T细胞的氧化损伤和凋亡,具有显著的光保护功效,其作用机制与增强细胞抗氧化能力、加速清除氧自由基以及促进胶原蛋白合成有关。
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 Deanship of Scientific Research from King Saud University
文摘The atmosphere protects humans,plants,animals,and microorganisms from damaging doses of ultraviolet-B(UVB) solar radiation(280-320 nm) because it modifies the UVB reaching the Earth's surface.This modification is a function of the solar radiation's path length through the atmosphere and the amount of each attenuator along the path length.The path length is determined by solar zenith angle(SZA).The present work explains the dependence of hemispherical transmittance of UVB on SZA.The database used consists of five years of hourly UVB and global solar radiation measurements.From 2001 to 2005,the South Valley University(SVU) meteorological research station(26.20°N,32.75°E) carried out these measurements on a horizontal surface.In addition,the corresponding extraterrestrial UVB(UVBext) and broadband solar radiation(Gext) were estimated.Consequently,the hemispherical transmittance of UVB(KtUVB) and the hemispherical transmittance of global solar radiation(Kt) were estimated.Furthermore,the UVB redaction due to the atmosphere was evaluated.An analysis of the dependence between KtUVB and SZA at different ranges of Kt was performed.A functional dependence between KtUVB and SZA(KtUVB=-a(SZA)+b) for very narrow Kt-ranges(width of ranges was 0.01) was developed.The results are discussed,and the sensitivity of △KtUVB to △SZA for very narrow Kt-ranges was studied.It was found that the sensitivity of △KtUVB to △SZA slightly increases with increased Kt,which means KtUVB is sensitive to SZA as Kt increases.The maximum correlation(R) between KtUVB and SZA was equal to-0.83 for Kt= 0.76.
文摘目的探讨窄谱中波紫外线(NB-UVB)家庭光疗与院内光疗治疗小腿斑块状银屑病的疗效比较。方法选择2018年7月至2020年6月嘉兴市第一医院皮肤科门诊收治的58例小腿斑块状银屑病患者,随机分为对照组28例和观察组30例收治的对照组给予外用卡泊三醇倍他米松软膏联合院内照射NB-UVB,观察组给予外用卡泊三醇倍他米松软膏联合家中自行照射NB-UVB,两组疗程均为8周。通过治疗前后银屑病皮损面积与严重程度指数(psoriasis area and severity index,PASI)评分判定疗效,记录皮肤不良反应发生情况,以调查问卷形式评价便利性。结果治疗后两组的PASI评分均较治疗前下降,差异有统计学意义(P<0.05);治疗后两组的PASI评分比较,差异无统计学意义(P>0.05)。对照组的总有效率为82.14%,观察组为76.67%,两组比较,差异无统计学意义(P>0.05)。对照组的轻度皮肤不良反应发生率为17.86%,观察组为13.33%,两组比较,差异无统计学意义(P>0.05)。观察组的便利性优于对照组,差异有统计学意义(P<0.05)。结论NB-UVB家庭光疗对比院内光疗治疗小腿斑块状银屑病,疗效相当,安全性良好,便利程度高,值得临床推广。
文摘Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.
