西罗莫司诱导自噬对UVB辐照下HaCaT细胞增殖和凋亡的初步影响
摘要
背景
中波紫外线主要作用于表皮,角质形成细胞是其重要的靶细胞,可引起角质形成细胞DNA损伤、氧化应激、细胞凋亡,从而造成皮肤光损伤。自噬是广泛存在于真核细胞的生理现象,可以被损伤、辐射和饥饿等应激因素诱导。白噬主要功能是降解变性蛋白质及受损细胞器,对于保持细胞内环境稳定极为重要,可以维持细胞在不利因素下存活。研究证实自噬在细胞损伤中被激活并起到一定的细胞保护作用。研究发现角质形成细胞也存在自噬,并可减轻角质形成细胞的炎症。角质形成细胞中也存在自噬现象,自噬在角质形成细胞UVB相关光损伤的角色尚不明确,西罗莫司作为自噬的诱导剂是否能减轻光损伤,我们对此做了相关初步探索。
目的
观察UVB辐照HaCaT细胞是否出现自噬现象；研究西罗莫司对UVB辐照下HaCaT细胞自噬水平的影响；检测西罗莫司对UVB辐照下HaCaT细胞增殖和凋亡的初步影响。
方法
以0、25、50mJ/cm2UVB三个剂量辐照HaCaT细胞,MDC染色法观察是否出现自噬体。设置6组：空白组、DMSO组、25mJ/cm2UVB组、25mJ/cm2UVB+西罗莫司组、50mJ/cm2UVB组、50mJ/cm2UVB+西罗莫司；MDC染色法观察并比较6组细胞出现自噬的水平；MTT法测定6组HaCaT细胞增殖能力；使用Hoechest染色法,AnnexinV-FITC、PI荧光标记细胞后流式细胞仪检测西罗莫司对UVB辐照下HaCaT细胞凋亡的影响。
结果
UVB照射HaCaT细胞后出现了自噬,0、25、50mJ/cm2UVB照射组自噬体阳性细胞百分率分别为1.07%±1.85%、9.37%±1.14%、16.29%±3.03%,三组间存在显著性差异(F=37.34,P<0.01)。
6组自噬体阳性细胞百分率分别为：0.56%0.79%、1.61%±2.27%、10.00%±0.47%、21.18%±3.03%、15.82%±4.13%、29.45%±1.24%,各组间均有显著性差异(F=45.23,P<0.01),且25mJ/cm2UVB+西罗莫司组、50mJ/cm2UVB+西罗莫司组的自噬水平均高于UVB组。
6组的细胞增殖抑制率分别为：0.01%±4.70%、8.40%±12.43%、9.43%±2.87%、6.22%±5.88%、12.12%±1.91%、14.17%±4.30%,组间有统计学意义(F=9.836,n=5,P<0.01);25mJ/cm2、50mJ/cm2UVB照射均抑制细胞增殖,25mJ/cm2UVB照射组抑制率为9.43%士2.87%,加入西罗莫司孵育后抑制率为6.22%士5.88%,减轻了抑制效应,而在50mJ/cm2UVB照射后加入西罗莫司,却增加了抑制率。
Hoechest染色法结果提示25mJ/cm2组、50mJ/cm2UVB组HaCaT细胞出现多数凋亡小体,50mJ/cm2组较25mJ/cm2组出现凋亡数量为多,加入西罗莫司干预后凋亡小体明显减少。
FCM结果提示6组早期凋亡百分比(LR)分别为：2.18%±1.30%、2.33%±1.96%、8.55%±8.44%、8.36%±0.70%、11.43%±11.81%、5.18%±5.83%,6组中晚期凋亡百分比(UR)分别为：1.72%±0.33%、2.15%±1.11%、1.72%±1.84%、12.48%±5.04%、14.24%±4.10%、8.97%±6.01%,50mJ/cm2UVB+西罗莫司组早期、中晚期凋亡百分比均低于50mJ/cm2UVB组,提示西罗莫司处理能减轻50mJ/cm2UVB辐射引起的细胞凋亡。
结论
UVB辐照HaCaT细胞出现自噬现象；西罗莫司能增加UVB辐照下HaCaT细胞的自噬水平；西罗莫司诱导自噬可减轻25mJ/cm2UVB辐照对HaCaT细胞增殖活性的抑制,而在50mJ/cm2UVB辐照下则加重对细胞的抑制。西罗莫司诱导自噬可减轻50mJ/cm2UVB辐照下HaCaT细胞的凋亡数量。西罗莫司诱导自噬对UVB辐照下HaCaT细胞光损伤有保护作用。
Introduction
Ultraviolet B (UVB) mainly targets keratinocytes of epidermis, resulting in photo relative skin injuries including DNA damage, oxidative stress and cell apoptosis. Autophagy,as a physiological phenomenon, extensively exists in eukaryotic cells. Autophagy can be triggered by radiation, hunger and other stresses. Autophagy performs a fundermental role of degeneration of denatured protains and injured organelles. Also, autophagy is of significance to maintain cell homeostasis and benefits cells survival undergoing inferior environment. Scientific researches have demonstrated that autophagy can be activated by cell injuries and autophagy supply a protective effect for host. Moreover, it is be suggested that autophagy exsits in keratinocytes and alleviates inflammation in keratinocytes. Whereas, it remains unclear whether antophagy will be induced in keratinocyte exposed to UVB. The role of autophagy is uncertain concerning damages in keratinocytes irradiated by UVB. Whether sirolimus inducing autophagy can alleviate UVB-injuries in keratinocytes is still unknown.Based on the above statements, we attempted to investigate the relative researches.
Objective
To detect whether autophagy can be induced by UVB irradiation in HaCaT cells. To explore the effects of sirolimus on autophagy level in UVB-irradiated HaCaT cells. To determine the effects of sirolimus on the proliferation of and the apoptosis in HaCaT cells irradiated by UVB.
Methods
HaCaT cells were irradiated by UVB containing doses of0、25、50mJ/cm2and followed by MDC staining to detect the autophagosome. Subsequently, HaCaT cells were divided into6groups as follows:untreated cells; cells treated with DMSO; cells irradiated by25mJ/cm2UVB; cells irradiated by25mJ/cm2UVB and treated with sirolimus; cells irradiated by50mJ/cm2UVB; cells irradiated by50mJ/cm2UVB and treated with sirolimus. Then the autophagosome in HaCaT cells of6groups was examined by MDC staining, and the percentages of the autophagosome positive cells of6groups were counted and compared. Furthermore, the proliferation of HaCaT cells of6groups were determined by MTT assay. The apoptosis in HaCaT cells of6groups were assessed by Hoechest staining and flow cytometry using AnnexinV-FITC and PI fluorescein.
Results
Autophagy was detected in HaCaT cells after UVB exposure. The percentages of the autophagosome positive cells in HaCaT cells receiving UVB irradiation containing doses of0,25,50mJ/cm2respectively were1.07%±1.85%,9.37%±1.14%,16.29%±3.03%, and were of significant difference (F=37.34, n=2,.P<0.01) among three UVB doses. Subsequently, the percentages of the autophagosome positive cells in HaCaT cells of6groups respectively were0.56%±0.79%,1.61%±2.27%,10.00%±0.47%,21.18%±3.03%,15.82%±4.13%,29.45%±1.24%, and similarly were of significant difference (F=45.23, n=5, P<0.01) among6groups. In details, the percentages of the autophagosome positive cells in HaCaT cells irradiated by UVB and sirolimus were higher than in cells irradiated by UVB only.
The inhibition ratio of proliferation of HaCaT cells of6groups respectively were0.01%±4.70%、8.40%±12.43%、9.43%±2.87%、6.22%±5.88%、12.12%±1.91%、14.17%±4.30%, and were different significantly (F=9.836, n=5,P<0.01). UVB irridiation containing doses of both25mJ/cm2and50mJ/cm2inhibited the proliferation of HaCaT cells. The ratio of suppression to cell proliferation of HaCaT cells irradiated by25mJ/cm2UVB was9.43%±2.87%, and was6.22%±5.88%of HaCaT cells irradiated by25mJ/cm2UVB and treated with sirolimus, suggesting that sirolimus relieved the suppression to proliferation of HaCaT cells irradiated by25mJ/cm2UVB. However, sirolimus aggravated inhibition ratio of proliferation of HaCaT cells irradiated by50mJ/cm2UVB.
Apoptotic cells and apoptotic bodies were examined obviously by Hoechest staining in groups of cells irradiated by25mJ/cm2or50mJ/cm2UVB, and the number of apoptotic cells and apoptotic bodies in groups of cells irradiated by50mJ/cm2UVB were more than in groups of cells irradiated by25mJ/cm2UVB. The number were decreased notabley in those groups of cells treated with sirolimus.
Data of flow cytometry using Annexin V-FITC and PI indicated that the percentages of prophase apoptotic cells in6groups respectively were2.18%±1.30%、2.33%±1.96%、 8.55%±8.44%、8.36%±0.70%、11.43%±1.81%、5.18%±5.83%, and the percentages of anaphase apoptotic cells in6groups respectively were1.72%±0.33%、2.15%±1.11%、1.72%±1.84%、12.48%±5.04%、14.24%±4.10%、8.97%±6.01%. Consistently, the percentages of both prophase apoptotic cells and anaphase apoptotic cells in groups treated with50mJ/cm2UVB and sirolimus were lower than treated with50mJ/cm2UVB only, allowing sirolimus reduce apoptotic cells and apoptotic bodies in HaCaT cells irradiated by50mJ/cm2UVB.
Conclusions
Autophagy occurred in HaCaT cells irradiated by UVB. The percentages of the autophagosome positive cells in HaCaT cells were improved by sirolimus. Sirolimus relieved the suppression to cell proliferation of cells irradiated by25mJ/cm2UVB. However, sirolimus aggravated the suppression to cell proliferation of cells irradiated by50mJ/cm2UVB. Sirolimus reduced apoptotic cells in HaCaT cells irradiated by50mJ/cm2UVB. Sirolimus inducing autophagy benefited the proliferation of and decreased the apoptosis cells in HaCaT Cells irradiated by UVB specificly.
中波紫外线主要作用于表皮,角质形成细胞是其重要的靶细胞,可引起角质形成细胞DNA损伤、氧化应激、细胞凋亡,从而造成皮肤光损伤。自噬是广泛存在于真核细胞的生理现象,可以被损伤、辐射和饥饿等应激因素诱导。白噬主要功能是降解变性蛋白质及受损细胞器,对于保持细胞内环境稳定极为重要,可以维持细胞在不利因素下存活。研究证实自噬在细胞损伤中被激活并起到一定的细胞保护作用。研究发现角质形成细胞也存在自噬,并可减轻角质形成细胞的炎症。角质形成细胞中也存在自噬现象,自噬在角质形成细胞UVB相关光损伤的角色尚不明确,西罗莫司作为自噬的诱导剂是否能减轻光损伤,我们对此做了相关初步探索。
目的
观察UVB辐照HaCaT细胞是否出现自噬现象；研究西罗莫司对UVB辐照下HaCaT细胞自噬水平的影响；检测西罗莫司对UVB辐照下HaCaT细胞增殖和凋亡的初步影响。
方法
以0、25、50mJ/cm2UVB三个剂量辐照HaCaT细胞,MDC染色法观察是否出现自噬体。设置6组：空白组、DMSO组、25mJ/cm2UVB组、25mJ/cm2UVB+西罗莫司组、50mJ/cm2UVB组、50mJ/cm2UVB+西罗莫司；MDC染色法观察并比较6组细胞出现自噬的水平；MTT法测定6组HaCaT细胞增殖能力；使用Hoechest染色法,AnnexinV-FITC、PI荧光标记细胞后流式细胞仪检测西罗莫司对UVB辐照下HaCaT细胞凋亡的影响。
结果
UVB照射HaCaT细胞后出现了自噬,0、25、50mJ/cm2UVB照射组自噬体阳性细胞百分率分别为1.07%±1.85%、9.37%±1.14%、16.29%±3.03%,三组间存在显著性差异(F=37.34,P<0.01)。
6组自噬体阳性细胞百分率分别为：0.56%0.79%、1.61%±2.27%、10.00%±0.47%、21.18%±3.03%、15.82%±4.13%、29.45%±1.24%,各组间均有显著性差异(F=45.23,P<0.01),且25mJ/cm2UVB+西罗莫司组、50mJ/cm2UVB+西罗莫司组的自噬水平均高于UVB组。
6组的细胞增殖抑制率分别为：0.01%±4.70%、8.40%±12.43%、9.43%±2.87%、6.22%±5.88%、12.12%±1.91%、14.17%±4.30%,组间有统计学意义(F=9.836,n=5,P<0.01);25mJ/cm2、50mJ/cm2UVB照射均抑制细胞增殖,25mJ/cm2UVB照射组抑制率为9.43%士2.87%,加入西罗莫司孵育后抑制率为6.22%士5.88%,减轻了抑制效应,而在50mJ/cm2UVB照射后加入西罗莫司,却增加了抑制率。
Hoechest染色法结果提示25mJ/cm2组、50mJ/cm2UVB组HaCaT细胞出现多数凋亡小体,50mJ/cm2组较25mJ/cm2组出现凋亡数量为多,加入西罗莫司干预后凋亡小体明显减少。
FCM结果提示6组早期凋亡百分比(LR)分别为：2.18%±1.30%、2.33%±1.96%、8.55%±8.44%、8.36%±0.70%、11.43%±11.81%、5.18%±5.83%,6组中晚期凋亡百分比(UR)分别为：1.72%±0.33%、2.15%±1.11%、1.72%±1.84%、12.48%±5.04%、14.24%±4.10%、8.97%±6.01%,50mJ/cm2UVB+西罗莫司组早期、中晚期凋亡百分比均低于50mJ/cm2UVB组,提示西罗莫司处理能减轻50mJ/cm2UVB辐射引起的细胞凋亡。
结论
UVB辐照HaCaT细胞出现自噬现象；西罗莫司能增加UVB辐照下HaCaT细胞的自噬水平；西罗莫司诱导自噬可减轻25mJ/cm2UVB辐照对HaCaT细胞增殖活性的抑制,而在50mJ/cm2UVB辐照下则加重对细胞的抑制。西罗莫司诱导自噬可减轻50mJ/cm2UVB辐照下HaCaT细胞的凋亡数量。西罗莫司诱导自噬对UVB辐照下HaCaT细胞光损伤有保护作用。
Introduction
Ultraviolet B (UVB) mainly targets keratinocytes of epidermis, resulting in photo relative skin injuries including DNA damage, oxidative stress and cell apoptosis. Autophagy,as a physiological phenomenon, extensively exists in eukaryotic cells. Autophagy can be triggered by radiation, hunger and other stresses. Autophagy performs a fundermental role of degeneration of denatured protains and injured organelles. Also, autophagy is of significance to maintain cell homeostasis and benefits cells survival undergoing inferior environment. Scientific researches have demonstrated that autophagy can be activated by cell injuries and autophagy supply a protective effect for host. Moreover, it is be suggested that autophagy exsits in keratinocytes and alleviates inflammation in keratinocytes. Whereas, it remains unclear whether antophagy will be induced in keratinocyte exposed to UVB. The role of autophagy is uncertain concerning damages in keratinocytes irradiated by UVB. Whether sirolimus inducing autophagy can alleviate UVB-injuries in keratinocytes is still unknown.Based on the above statements, we attempted to investigate the relative researches.
Objective
To detect whether autophagy can be induced by UVB irradiation in HaCaT cells. To explore the effects of sirolimus on autophagy level in UVB-irradiated HaCaT cells. To determine the effects of sirolimus on the proliferation of and the apoptosis in HaCaT cells irradiated by UVB.
Methods
HaCaT cells were irradiated by UVB containing doses of0、25、50mJ/cm2and followed by MDC staining to detect the autophagosome. Subsequently, HaCaT cells were divided into6groups as follows:untreated cells; cells treated with DMSO; cells irradiated by25mJ/cm2UVB; cells irradiated by25mJ/cm2UVB and treated with sirolimus; cells irradiated by50mJ/cm2UVB; cells irradiated by50mJ/cm2UVB and treated with sirolimus. Then the autophagosome in HaCaT cells of6groups was examined by MDC staining, and the percentages of the autophagosome positive cells of6groups were counted and compared. Furthermore, the proliferation of HaCaT cells of6groups were determined by MTT assay. The apoptosis in HaCaT cells of6groups were assessed by Hoechest staining and flow cytometry using AnnexinV-FITC and PI fluorescein.
Results
Autophagy was detected in HaCaT cells after UVB exposure. The percentages of the autophagosome positive cells in HaCaT cells receiving UVB irradiation containing doses of0,25,50mJ/cm2respectively were1.07%±1.85%,9.37%±1.14%,16.29%±3.03%, and were of significant difference (F=37.34, n=2,.P<0.01) among three UVB doses. Subsequently, the percentages of the autophagosome positive cells in HaCaT cells of6groups respectively were0.56%±0.79%,1.61%±2.27%,10.00%±0.47%,21.18%±3.03%,15.82%±4.13%,29.45%±1.24%, and similarly were of significant difference (F=45.23, n=5, P<0.01) among6groups. In details, the percentages of the autophagosome positive cells in HaCaT cells irradiated by UVB and sirolimus were higher than in cells irradiated by UVB only.
The inhibition ratio of proliferation of HaCaT cells of6groups respectively were0.01%±4.70%、8.40%±12.43%、9.43%±2.87%、6.22%±5.88%、12.12%±1.91%、14.17%±4.30%, and were different significantly (F=9.836, n=5,P<0.01). UVB irridiation containing doses of both25mJ/cm2and50mJ/cm2inhibited the proliferation of HaCaT cells. The ratio of suppression to cell proliferation of HaCaT cells irradiated by25mJ/cm2UVB was9.43%±2.87%, and was6.22%±5.88%of HaCaT cells irradiated by25mJ/cm2UVB and treated with sirolimus, suggesting that sirolimus relieved the suppression to proliferation of HaCaT cells irradiated by25mJ/cm2UVB. However, sirolimus aggravated inhibition ratio of proliferation of HaCaT cells irradiated by50mJ/cm2UVB.
Apoptotic cells and apoptotic bodies were examined obviously by Hoechest staining in groups of cells irradiated by25mJ/cm2or50mJ/cm2UVB, and the number of apoptotic cells and apoptotic bodies in groups of cells irradiated by50mJ/cm2UVB were more than in groups of cells irradiated by25mJ/cm2UVB. The number were decreased notabley in those groups of cells treated with sirolimus.
Data of flow cytometry using Annexin V-FITC and PI indicated that the percentages of prophase apoptotic cells in6groups respectively were2.18%±1.30%、2.33%±1.96%、 8.55%±8.44%、8.36%±0.70%、11.43%±1.81%、5.18%±5.83%, and the percentages of anaphase apoptotic cells in6groups respectively were1.72%±0.33%、2.15%±1.11%、1.72%±1.84%、12.48%±5.04%、14.24%±4.10%、8.97%±6.01%. Consistently, the percentages of both prophase apoptotic cells and anaphase apoptotic cells in groups treated with50mJ/cm2UVB and sirolimus were lower than treated with50mJ/cm2UVB only, allowing sirolimus reduce apoptotic cells and apoptotic bodies in HaCaT cells irradiated by50mJ/cm2UVB.
Conclusions
Autophagy occurred in HaCaT cells irradiated by UVB. The percentages of the autophagosome positive cells in HaCaT cells were improved by sirolimus. Sirolimus relieved the suppression to cell proliferation of cells irradiated by25mJ/cm2UVB. However, sirolimus aggravated the suppression to cell proliferation of cells irradiated by50mJ/cm2UVB. Sirolimus reduced apoptotic cells in HaCaT cells irradiated by50mJ/cm2UVB. Sirolimus inducing autophagy benefited the proliferation of and decreased the apoptosis cells in HaCaT Cells irradiated by UVB specificly.
引文
[1]赵辨.中国临床皮肤病学[M].南京：江苏科学技术出版社,2010：165-170.
[2]顾恒,常宝珠,陈崑.光皮肤病学[M].北京：人民军医出版社,2009：14-33.
[3]Guo W, An Y, Jiang L, et al. The protective effects of hydroxytyrosol against UVB-induced DNA damage in HaCaT cells[J]. Phytother Res,2010,24(3):352-359.
[4]Vostalova J, Zdarilova A, Svobodova A. Prunella vulgaris extract and rosmarinic acid prevent UVB-induced DNA damage and oxidative stress in HaCaT keratinocytes[J]. Arch Dermatol Res,2010,302(3):171-181.
[5]Svobodova A, Zdarilova A, Vostalova J. Lonicera caerulea and Vaccinium myrtillus fruit polyphenols protect HaCaT keratinocytes against UVB-induced phototoxic stress and DNA damage[J]. J Dermatol Sci,2009,56(3):196-204.
[6]Rabinowitz J D, White E. Autophagy and metabolism [J]. Science,2010,330(6009): 1344-1348.
[7]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[J]. Exp Dermatol,2011,20(3):263-268.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J], J Immunol,2011, 186(2):1248-1258.
[9]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes[J]. Mol Carcinog,2012,51 (3):268-279.
[10]Mathew R, Karp C M, Beaudoin B, et al. Autophagy suppresses tumorigenesis through elimination of p62[J]. Cell,2009,137(6):1062-1075.
[11]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[12]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[13]Sarkar S, Ravikumar B, Floto R A, et al. Rapamycin and mTOR-independent autophagy inducers ameliorate toxicity of polyglutamine-expanded huntingtin and related proteinopathies[J]. Cell Death Differ,2009,16(1):46-56.
[14]Klionsky D J. Crohn disease and autophagy[J]. Autophagy,2009,5(2):139.
[15]Kim H P, Wang X, Chen Z H, et al. Autophagic proteins regulate cigarette smoke-induced apoptosis:protective role of heme oxygenase-1[J]. Autophagy,2008, 4(7):887-895.
[16]Wu Y T, Tan H L, Huang Q, et al. Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy[J]. Autophagy,2009, 5(6):824-834.
[17]何维.医学免疫学[M].北京：人民卫生出版社,2010：405.
[18]陈新谦,金有豫,汤光.新编药物学[M.北京：人民卫生出版社,2011：766.
[19]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[20]Brazelton T R, Morris R E. Molecular mechanisms of action of new xenobiotic immunosuppressive drugs:tacrolimus (FK506), sirolimus (rapamycin), mycophenolate mofetil and leflunomide[J]. Curr Opin Immunol,1996,8(5):710-720.
[21]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[22]Jaboin J J, Shinohara E T, Moretti L, et al. The role of mTOR inhibition in augmenting radiation induced autophagy [J]. Technol Cancer Res Treat,2007, 6(5):443-447.
[23]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[1]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[2]张青松,顾恒.UvB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[31陈旭.紫外线致皮肤成纤维细胞和角质形成细胞中的自噬研究[D].北京协和医学院,2012.
[4]Yang Y, Wang H, Wang S, et al. GSK3beta signaling is involved in ultraviolet B-induced activation of autophagy in epidermal cells[J]. Int J Oncol,2012,41(5): 1782-1788.
[5]赵辨.中国临床皮肤病学[M].南京：江苏科学技术出版社,2010：165-170.
[6]Guo Z, Zhou B, Liu W, et al. MiR-23a regulates DNA damage repair and apoptosis in UVB-irradiated HaCaT cells[J]. J Dermatol Sci,2013,69(1):68-76.
[7]Lee C, Park G H, Ahn E M, et al. Protective effect of Codium fragile against UVB-induced pro-inflammatory and oxidative damages in HaCaT cells and BALB/c mice[J]. Fitoterapia,2013,86C:54-63.
[8]Vostalova J, Zdarilova A, Svobodova A. Prunella vulgaris extract and rosmarinic acid prevent UVB-induced DNA damage and oxidative stress in HaCaT keratinocytes[J]. Arch Dermatol Res,2010,302(3):171-181.
[9]Svobodova A, Zdarilova A, Vostalova J. Lonicera caerulea and Vaccinium myrtillus fruit polyphenols protect HaCaT keratinocytes against UVB-induced phototoxic stress and DNA damage[J]. J Dermatol Sci,2009,56(3):196-204.
[10]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[11]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[j]. Exp Dermatol,2011,20(3):263-268.
[12]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[13]Takasawa R, Nakamura H, Mori T, et al. Differential apoptotic pathways in human keratinocyte HaCaT cells exposed to UVB and UVC[J]. Apoptosis,2005,10(5): 1121-1130.
[14]Wang Q, Liu W, Zeng H, et al. p53-mediated autophagy adjustment is involved in the protection of silibinin against murine dermal inflammation and epidermal apoptosis induced by UVB irradiation[J]. J Asian Nat Prod Res,2013,15(2):117-129.
[15]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes[J]. Mol Carcinog,2012,51(3):268-279.
[1]Kim H P, Wang X, Chen Z H, et al. Autophagic proteins regulate cigarette smoke-induced apoptosis:protective role of heme oxygenase-1[J]. Autophagy,2008, 4(7):887-895.
[2]Qiang L, Wu C, Ming M, et al. Autophagy controls p38 activation to promote cell survival under genotoxic stress[J]. J Biol Chem,2013,288(3):1603-1611.
[3]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[4]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[5]秦正红,乐卫东.自噬：生物学与疾病[M].北京：科学出版社,2011：60-61.
[6]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[7]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[J]. Exp Dermatol,2011,20(3):263-268.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[9]任发亮,黄丹,陈旭,等.角质形成细胞中的自噬现象[J].国际皮肤性病学杂志,2012,38(5)：329-332.
[10]Jaboin J J, Shinohara E T, Moretti L, et al. The role of mTOR inhibition in augmenting radiation induced autophagy [J]. Technol Cancer Res Treat,2007,6(5): 443-447.
[11]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[12]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[13]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[1]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[2]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[3]秦正红,乐卫东.自噬：生物学与疾病[[.北京：科学出版社,2011：60-61.
[4]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[5]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[6]Massey D C, Bredin F, Parkes M. Use of sirolimus (rapamycin) to treat refractory Crohn's disease[J]. Gut,2008,57(9):1294-1296.
[7]Cleary C, Linde J A, Hiscock K M, et al. Antidepressive-like effects of rapamycin in animal models:Implications for mTOR inhibition as a new target for treatment of affective disorders[J]. Brain Res Bull,2008,76(5):469-473.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[9]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[10]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[11]Sudarsanam S, Johnson D E. Functional consequences of mTOR inhibition[J], Curr Opin Drug Discov Devel,2010,13(1):31-40.
[12]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[13]Halapas A, Armakolas A, Koutsilieris M. Autophagy:a target for therapeutic interventions in myocardial pathophysiology[J]. Expert Opin Ther Targets,2008,12(12): 1509-1522.
[1]秦正红,乐卫东.自噬：生物学与疾病[M].北京：科学出版社,2011：60-61.
[2]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[3]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[4]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[5]Takasawa R, Nakamura H, Mori T, et al. Differential apoptotic pathways in human keratinocyte HaCaT cells exposed to UVB and UVC[J]. Apoptosis,2005,10(5): 1121-1130.
[6]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[7]Sarkar S, Ravikumar B, Floto R A, et al. Rapamycin and mTOR-independent autophagy inducers ameliorate toxicity of polyglutamine-expanded huntingtin and related proteinopathies[J], Cell Death Differ,2009,16(1):46-56.
[8]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[9]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[10]Halapas A, Armakolas A, Koutsilieris M. Autophagy:a target for therapeutic interventions in myocardial pathophysiology[J]. Expert Opin Ther Targets,2008,12(12): 1509-1522.
[11]Wu Y T, Tan H L, Huang Q, et al. Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy [J]. Autophagy,2009, 5(6):824-834.
[12]Klionsky D J. Crohn disease and autophagy[J]. Autophagy,2009,5(2):139.
[13]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[14]Qiang L, Wu C, Ming M, et al. Autophagy controls p38 activation to promote cell survival undergenotoxic stress[J]. J Biol Chem,2013,288(3):1603-1611.
[15]Wang Q, Liu W, Zeng H, et al. p53-mediated autophagy adjustment is involved in the protection of silibinin against murine dermal inflammation and epidermal apoptosis induced by UVB irradiation[J]. J Asian Nat Prod Res,2013,15(2):117-129.
[1]Rabinowitz J D, White E. Autophagy and metabolism. Science,2010,330(6009): 1344-1348.
[2]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?. Exp Dermatol,2011,20(3):263-268.
[3]Chatterjea S M, Resing K A, Old W, et al. Optimization of filaggrin expression and processing in cultured rat keratinocytes. J Dermatol Sci,2011,61(1):51-59.
[4]Deruy E, Gosselin K, Vercamer C, et al. MnSOD upregulation induces autophagic programmed cell death in senescent keratinocytes. PLoS One,2010,5(9):e12712.
[5]Gosselin K, Deruy E, Martien S, et al. Senescent keratinocytes die by autophagic programmed cell death[J]. Am J Pathol,2009,174(2):423-435.
[6]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes. Mol Carcinog, 2012,51(3):268-279.
[7]Wu Y N, Yang L X, Shi X Y, et al. The selective growth inhibition of oral cancer by iron core-gold shell nanoparticles through mitochondria-mediated autophagy. Biomaterials,2011,32(20):4565-4573.
[8]Claerhout S, Verschooten L, Van Kelst S, et al. Concomitant inhibition of AKT and autophagy is required for efficient cisplatin-induced apoptosis of metastatic skin carcinoma. Int J Cancer,2010,127(12):2790-2803.
[9]Ratovitski E A. DeltaNp63alpha/IRF6 interplay activates NOS2 transcription and induces autophagy upon tobacco exposure.Arch Biochem Biophys,2011,506(2): 208-215.
[10]Zhou X, Munger K. Expression of the human papillomavirus type 16 E7 oncoprotein induces an autophagy-related process and sensitizes normal human keratinocytes to cell death in response to growth factor deprivation. Virology,2009, 385(1):192-197.
[11]Nomura H, Uzawa K, Yamano Y, et al. Overexpression and altered subcellular localization of autophagy-related 16-like 1 in human oral squamous-cell carcinoma: correlation with lymphovascular invasion and lymph-node metastasis. Hum Pathol,2009, 40(1):83-91.
[12]Wang R C, Levine B. Calcipotriol induces autophagy in HeLa cells and keratinocytes. J Invest Dermatol,2011,131(4):990-993.
[13]Silva J N, Galmiche A, Tome J P, et al. Chain-dependent photocytotoxicity of tricationic porphyrin conjugates and related mechanisms of cell death in proliferating human skin keratinocytes. Biochem Pharmacol,2010,80(9):1373-1385.
[14]von Bultzingslowen I, Jontell M, Hurst P, et al.5-Fluorouracil induces autophagic degeneration in rat oral keratinocytes. Oral Oncol,2001,37(6):537-544.
[15]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1. J Immunol,2011,186(2): 1248-1258.
[16]Nakatani T, Beitner H. A transmission electron microscopic study of macromelanosomes in sporadic dysplastic naevi. Okajimas Folia Anat Jpn,1992,69(2-3): 109-114.
[17]Hirone T, Eryu Y. Ultrastructure of giant pigment granules in lentigo simplex. Acta Derm Venereol,1978,58(3):223-229.
[1]Harman K E, Albert S, Black M M. Guidelines for the management of pemphigus vulgaris[J]. Br J Dermatol;2003,149(5):926-937.
[2]Hashimoto T. Treatment strategies for pemphigus vulgaris in Japan[J]. Expert Opin Pharmacother,2008,9(9):1519-1530.
[3]Fox L P, Pandya A G. Pulse intravenous cyclophosphamide therapy for dermatologic disorders[J]. Dermatol Clin,2000,18(3):459-473.
[4]Olszewska M, Kolacinska-Strasz Z, Sulej J, et al. Efficacy and safety of cyclophosphamide, azathioprine, and cyclosporine (ciclosporin) as adjuvant drugs in pemphigus vulgaris[J]. Am J Clin Dermatol,2007,8(2):85-92.
[5]Cummins D L, Mimouni D, Anhalt G J, et al. Oral cyclophosphamide for treatment of pemphigus vulgaris and foliaceus[J]. J Am Acad Dermatol,2003,49(2):276-280.
[6]Pasricha J S, Khaitan B K, Raman R S, et al. Dexamethasone-cyclophosphamide pulse therapy for pemphigus[J]. Int J Dermatol,1995,34(12):875-882.
[7]Pandya A G, Sontheimer R D. Treatment of pemphigus vulgaris with pulse intravenous cyclophosphamide[J]. Arch Dermatol,1992,128(12):1626-1630.
[8]Momeni A Z, Iraji F, Aminjavaheri M, et al. The use of oral cyclophosphamide with dexamethasone pulse therapy in the treatment of pemphigus vulgaris[J]. J Dermatolog Treat,2007,18(5):275-278.
[9]Hayag M V, Cohen J A, Kerdel F A. Immunoablative high-dose cyclophosphamide without stem cell rescue in a patient with pemphigus vulgaris[J]. J Am Acad Dermatol,2000,43(6):1065-1069.
[10]Espana A A, Panizo C, Fernandez S, et al. [Prolonged complete clinical remission in patients with severe pemphigus vulgaris after cycles of intravenous cyclophosphamide][J]. Actas Dermosifiliogr,2009,100(2):113-120.
[11]Kanwar A J, De D. Pemphigus in India[J]. Indian J Dermatol Venereol Leprol,2011, 77(4):439-449.
[12]Tirado-Sanchez A, Leon-Dorantes G. Treatment of pemphigus vulgaris. An overview in Mexico[J]. Allergol Immunopathol (Madr),2006,34(1):10-16.
[13]Kanwar A J, Kaur S, Thami G P. Long-term efficacy of dexamethasone-cyclophosphamide pulse therapy in pemphigus [J]. Dermatology,2002, 204(3):228-231.
[14]Martin L K, Werth V P, Villaneuva E V, et al. A systematic review of randomized controlled trials for pemphigus vulgaris and pemphigus foliaceus[J]. J Am Acad Dermatol,2011,64(5):903-908.
[15]Martin L K, Werth V, Villanueva E, et al. Interventions for pemphigus vulgaris and pemphigus foliaceus[J]. Cochrane Database Syst Rev,2009(1):D6263.
[16]Scully C, Challacombe S J. Pemphigus vulgaris:update on etiopathogenesis, oral manifestations, and management[J]. Crit Rev Oral Biol Med,2002,13(5):397-408.
[17]Yeh S W, Sami N, Ahmed R A. Treatment of pemphigus vulgaris:current and emerging options[J]. Am J Clin Dermatol,2005,6(5):327-342.
[18]Kaur S, Kanwar A J. Dexamethasone-cyclophosphamide pulse therapy in pemphigus[J]. Int J Dermatol,1990,29(5):371-374.
[19]Sethy P K, Khandpur S, Sharma V K. Randomized open comparative trial of dexamethasone-cyclophosphamide pulse and daily oral cyclophosphamide versus cyclophosphamide pulse and daily oral prednisolone in pemphigus vulgarisfJ]. Indian J Dermatol Venereol Leprol,2009,75(5):476-482.
[20]邓一文.治疗天疱疮的循证医学研究进展[J].临床口腔医学杂志,2010,26(11)：692-696.
[21]李薇,熊琳,卫茂玲.我国天疱疮治疗性研究的文献评价[J].华西医学,2005,20(4)：624-625.
[22]Mignogna M D.Lo M L, Mignogna R E, et al. Oral pemphigus:long term behaviour and clinical response to treatment with deflazacort in sixteen cases[J]. J Oral Pathol Med,2000,29(4):145-152.
[23]Jain R, Kumar B. Immediate and delayed complications of dexamethasone cyclophosphamide pulse (DCP) therapy[J]. J Dermatol,2003,30(10):713-718.
[24]Mabrouk D, Gurcan H M, Keskin D B, et al. Association between cancer and immunosuppressive therapy--analysis of selected studies in pemphigus and pemphigoid[J]. Ann Pharmacother,2010,44(11):1770-1776.
[25]Zivanovic D, Medenica L, Tanasilovic S, et al. Dexamethasone-cyclophosphamide pulse therapy in pemphigus:a review of 72 cases[J]. Am J Clin Dermatol,2010,11(2): 123-129.
[26]Ramam M. Prolonged antimicrobial and oral cyclophosphamide therapy in pemphigus:need for caution[J]. Indian J Dermatol Venereol Leprol,2009,75(1):85.
[1]Hanifin J M, Cooper K D, Ho V C, et al. Guidelines of care for atopic dermatitis, developed in accordance with the American Academy of Dermatology (AAD)/American Academy of Dermatology Association "Administrative Regulations for Evidence-Based Clinical Practice Guidelines"[J]. J Am Acad Dermatol,2004,50(3):391-404.
[2]Akdis C A, Akdis M, Bieber T, et al. Diagnosis and treatment of atopic dermatitis in children and adults:European Academy of Allergology and Clinical Immunology/American Academy of Allergy, Asthma and Immunology/PRACTALL Consensus Report[J]. Allergy,2006,61(8):969-987.
[3]中华医学会皮肤性病学分会免疫学组.中国特应性皮炎诊断和治疗指南[J].中华皮肤科杂志,2008,41(11)：772-773.
[4]Saeki H, Furue M, Furukawa F, et al. Guidelines for management of atopic dermatitis[J]. J Dermatol,2009,36(10):563-577.
[5]Darsow U, Wollenberg A, Simon D, et al. ETFAD/EADV eczema task force 2009 position paper on diagnosis and treatment of atopic dermatitis[J]. J Eur Acad Dermatol Venereol,2010,24(3):317-328.
[6]Ring J, Alomar A, Bieber T, et al. Guidelines for Treatment of Atopic Eczema (Atopic Dermatitis). For the European Dermatology Forum (EDF), and the European Academy of Dermato-Venereology (EADV), and European Federation of Allergy (EFA), and the European Society of Pediatric Dermatology (ESPD).2011.UD/JR,26.04.2011.
[7]顾恒,陈祥生,陈崑.特应性皮炎诊断标准的评价[J].中华皮肤科杂志,2000,33(4)：222.
[8]Gu H, Chen X S, Chen K, et al. Evaluation of diagnostic criteria for atopic dermatitis: validity of the criteria of Williams et al. in a hospital-based setting[J]. Br J Dermatol,2001,145(3):428-433.
[9]Severity scoring of atopic dermatitis:the SCORAD index. Consensus Report of the European Task Force on Atopic Dermatitis[J]. Dermatology,1993,186(1):23-31.
[2]顾恒,常宝珠,陈崑.光皮肤病学[M].北京：人民军医出版社,2009：14-33.
[3]Guo W, An Y, Jiang L, et al. The protective effects of hydroxytyrosol against UVB-induced DNA damage in HaCaT cells[J]. Phytother Res,2010,24(3):352-359.
[4]Vostalova J, Zdarilova A, Svobodova A. Prunella vulgaris extract and rosmarinic acid prevent UVB-induced DNA damage and oxidative stress in HaCaT keratinocytes[J]. Arch Dermatol Res,2010,302(3):171-181.
[5]Svobodova A, Zdarilova A, Vostalova J. Lonicera caerulea and Vaccinium myrtillus fruit polyphenols protect HaCaT keratinocytes against UVB-induced phototoxic stress and DNA damage[J]. J Dermatol Sci,2009,56(3):196-204.
[6]Rabinowitz J D, White E. Autophagy and metabolism [J]. Science,2010,330(6009): 1344-1348.
[7]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[J]. Exp Dermatol,2011,20(3):263-268.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J], J Immunol,2011, 186(2):1248-1258.
[9]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes[J]. Mol Carcinog,2012,51 (3):268-279.
[10]Mathew R, Karp C M, Beaudoin B, et al. Autophagy suppresses tumorigenesis through elimination of p62[J]. Cell,2009,137(6):1062-1075.
[11]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[12]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[13]Sarkar S, Ravikumar B, Floto R A, et al. Rapamycin and mTOR-independent autophagy inducers ameliorate toxicity of polyglutamine-expanded huntingtin and related proteinopathies[J]. Cell Death Differ,2009,16(1):46-56.
[14]Klionsky D J. Crohn disease and autophagy[J]. Autophagy,2009,5(2):139.
[15]Kim H P, Wang X, Chen Z H, et al. Autophagic proteins regulate cigarette smoke-induced apoptosis:protective role of heme oxygenase-1[J]. Autophagy,2008, 4(7):887-895.
[16]Wu Y T, Tan H L, Huang Q, et al. Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy[J]. Autophagy,2009, 5(6):824-834.
[17]何维.医学免疫学[M].北京：人民卫生出版社,2010：405.
[18]陈新谦,金有豫,汤光.新编药物学[M.北京：人民卫生出版社,2011：766.
[19]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[20]Brazelton T R, Morris R E. Molecular mechanisms of action of new xenobiotic immunosuppressive drugs:tacrolimus (FK506), sirolimus (rapamycin), mycophenolate mofetil and leflunomide[J]. Curr Opin Immunol,1996,8(5):710-720.
[21]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[22]Jaboin J J, Shinohara E T, Moretti L, et al. The role of mTOR inhibition in augmenting radiation induced autophagy [J]. Technol Cancer Res Treat,2007, 6(5):443-447.
[23]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[1]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[2]张青松,顾恒.UvB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[31陈旭.紫外线致皮肤成纤维细胞和角质形成细胞中的自噬研究[D].北京协和医学院,2012.
[4]Yang Y, Wang H, Wang S, et al. GSK3beta signaling is involved in ultraviolet B-induced activation of autophagy in epidermal cells[J]. Int J Oncol,2012,41(5): 1782-1788.
[5]赵辨.中国临床皮肤病学[M].南京：江苏科学技术出版社,2010：165-170.
[6]Guo Z, Zhou B, Liu W, et al. MiR-23a regulates DNA damage repair and apoptosis in UVB-irradiated HaCaT cells[J]. J Dermatol Sci,2013,69(1):68-76.
[7]Lee C, Park G H, Ahn E M, et al. Protective effect of Codium fragile against UVB-induced pro-inflammatory and oxidative damages in HaCaT cells and BALB/c mice[J]. Fitoterapia,2013,86C:54-63.
[8]Vostalova J, Zdarilova A, Svobodova A. Prunella vulgaris extract and rosmarinic acid prevent UVB-induced DNA damage and oxidative stress in HaCaT keratinocytes[J]. Arch Dermatol Res,2010,302(3):171-181.
[9]Svobodova A, Zdarilova A, Vostalova J. Lonicera caerulea and Vaccinium myrtillus fruit polyphenols protect HaCaT keratinocytes against UVB-induced phototoxic stress and DNA damage[J]. J Dermatol Sci,2009,56(3):196-204.
[10]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[11]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[j]. Exp Dermatol,2011,20(3):263-268.
[12]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[13]Takasawa R, Nakamura H, Mori T, et al. Differential apoptotic pathways in human keratinocyte HaCaT cells exposed to UVB and UVC[J]. Apoptosis,2005,10(5): 1121-1130.
[14]Wang Q, Liu W, Zeng H, et al. p53-mediated autophagy adjustment is involved in the protection of silibinin against murine dermal inflammation and epidermal apoptosis induced by UVB irradiation[J]. J Asian Nat Prod Res,2013,15(2):117-129.
[15]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes[J]. Mol Carcinog,2012,51(3):268-279.
[1]Kim H P, Wang X, Chen Z H, et al. Autophagic proteins regulate cigarette smoke-induced apoptosis:protective role of heme oxygenase-1[J]. Autophagy,2008, 4(7):887-895.
[2]Qiang L, Wu C, Ming M, et al. Autophagy controls p38 activation to promote cell survival under genotoxic stress[J]. J Biol Chem,2013,288(3):1603-1611.
[3]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[4]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[5]秦正红,乐卫东.自噬：生物学与疾病[M].北京：科学出版社,2011：60-61.
[6]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[7]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?[J]. Exp Dermatol,2011,20(3):263-268.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[9]任发亮,黄丹,陈旭,等.角质形成细胞中的自噬现象[J].国际皮肤性病学杂志,2012,38(5)：329-332.
[10]Jaboin J J, Shinohara E T, Moretti L, et al. The role of mTOR inhibition in augmenting radiation induced autophagy [J]. Technol Cancer Res Treat,2007,6(5): 443-447.
[11]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[12]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[13]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[1]张青松,顾恒.UVB对人皮肤成纤维细胞自噬影响的初步研究[J].中国麻风皮肤病杂志,2008,24(7)：511-513.
[2]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[3]秦正红,乐卫东.自噬：生物学与疾病[[.北京：科学出版社,2011：60-61.
[4]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[5]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[6]Massey D C, Bredin F, Parkes M. Use of sirolimus (rapamycin) to treat refractory Crohn's disease[J]. Gut,2008,57(9):1294-1296.
[7]Cleary C, Linde J A, Hiscock K M, et al. Antidepressive-like effects of rapamycin in animal models:Implications for mTOR inhibition as a new target for treatment of affective disorders[J]. Brain Res Bull,2008,76(5):469-473.
[8]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1[J]. J Immunol,2011, 186(2):1248-1258.
[9]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[10]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[11]Sudarsanam S, Johnson D E. Functional consequences of mTOR inhibition[J], Curr Opin Drug Discov Devel,2010,13(1):31-40.
[12]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[13]Halapas A, Armakolas A, Koutsilieris M. Autophagy:a target for therapeutic interventions in myocardial pathophysiology[J]. Expert Opin Ther Targets,2008,12(12): 1509-1522.
[1]秦正红,乐卫东.自噬：生物学与疾病[M].北京：科学出版社,2011：60-61.
[2]Rabinowitz J D, White E. Autophagy and metabolism[J]. Science,2010,330(6009): 1344-1348.
[3]Hung S Y, Huang W P, Liou H C, et al. Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy,2009,5(4):502-510.
[4]陈旭,张青松,鞠梅,等.中波紫外线诱导人皮肤成纤维细胞自噬对凋亡影响的初步研究[J].中华皮肤科杂志,2012,45(7)：496-500.
[5]Takasawa R, Nakamura H, Mori T, et al. Differential apoptotic pathways in human keratinocyte HaCaT cells exposed to UVB and UVC[J]. Apoptosis,2005,10(5): 1121-1130.
[6]Patsenker E, Schneider V, Ledermann M, et al. Potent antifibrotic activity of mTOR inhibitors sirolimus and everolimus but not of cyclosporine A and tacrolimus in experimental liver fibrosis[J]. J Hepatol,2011.55(2):p.388-98.
[7]Sarkar S, Ravikumar B, Floto R A, et al. Rapamycin and mTOR-independent autophagy inducers ameliorate toxicity of polyglutamine-expanded huntingtin and related proteinopathies[J], Cell Death Differ,2009,16(1):46-56.
[8]Jung C H, Ro S H, Cao J, et al. mTOR regulation of autophagy[J]. FEBS Lett,2010,584(7):1287-1295.
[9]Hayashi S, Yamamoto A, You F, et al. The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy[J]. Am J Pathol,2009,175(5):2226-2234.
[10]Halapas A, Armakolas A, Koutsilieris M. Autophagy:a target for therapeutic interventions in myocardial pathophysiology[J]. Expert Opin Ther Targets,2008,12(12): 1509-1522.
[11]Wu Y T, Tan H L, Huang Q, et al. Activation of the PI3K-Akt-mTOR signaling pathway promotes necrotic cell death via suppression of autophagy [J]. Autophagy,2009, 5(6):824-834.
[12]Klionsky D J. Crohn disease and autophagy[J]. Autophagy,2009,5(2):139.
[13]Herrero-Martin G, Hoyer-Hansen M, Garcia-Garcia C, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells[J]. EMBO J,2009,28(6):677-685.
[14]Qiang L, Wu C, Ming M, et al. Autophagy controls p38 activation to promote cell survival undergenotoxic stress[J]. J Biol Chem,2013,288(3):1603-1611.
[15]Wang Q, Liu W, Zeng H, et al. p53-mediated autophagy adjustment is involved in the protection of silibinin against murine dermal inflammation and epidermal apoptosis induced by UVB irradiation[J]. J Asian Nat Prod Res,2013,15(2):117-129.
[1]Rabinowitz J D, White E. Autophagy and metabolism. Science,2010,330(6009): 1344-1348.
[2]Aymard E, Barruche V, Naves T, et al. Autophagy in human keratinocytes:an early step of the differentiation?. Exp Dermatol,2011,20(3):263-268.
[3]Chatterjea S M, Resing K A, Old W, et al. Optimization of filaggrin expression and processing in cultured rat keratinocytes. J Dermatol Sci,2011,61(1):51-59.
[4]Deruy E, Gosselin K, Vercamer C, et al. MnSOD upregulation induces autophagic programmed cell death in senescent keratinocytes. PLoS One,2010,5(9):e12712.
[5]Gosselin K, Deruy E, Martien S, et al. Senescent keratinocytes die by autophagic programmed cell death[J]. Am J Pathol,2009,174(2):423-435.
[6]Tong X, Smith K A, Pelling J C. Apigenin, a chemopreventive bioflavonoid, induces AMP-activated protein kinase activation in human keratinocytes. Mol Carcinog, 2012,51(3):268-279.
[7]Wu Y N, Yang L X, Shi X Y, et al. The selective growth inhibition of oral cancer by iron core-gold shell nanoparticles through mitochondria-mediated autophagy. Biomaterials,2011,32(20):4565-4573.
[8]Claerhout S, Verschooten L, Van Kelst S, et al. Concomitant inhibition of AKT and autophagy is required for efficient cisplatin-induced apoptosis of metastatic skin carcinoma. Int J Cancer,2010,127(12):2790-2803.
[9]Ratovitski E A. DeltaNp63alpha/IRF6 interplay activates NOS2 transcription and induces autophagy upon tobacco exposure.Arch Biochem Biophys,2011,506(2): 208-215.
[10]Zhou X, Munger K. Expression of the human papillomavirus type 16 E7 oncoprotein induces an autophagy-related process and sensitizes normal human keratinocytes to cell death in response to growth factor deprivation. Virology,2009, 385(1):192-197.
[11]Nomura H, Uzawa K, Yamano Y, et al. Overexpression and altered subcellular localization of autophagy-related 16-like 1 in human oral squamous-cell carcinoma: correlation with lymphovascular invasion and lymph-node metastasis. Hum Pathol,2009, 40(1):83-91.
[12]Wang R C, Levine B. Calcipotriol induces autophagy in HeLa cells and keratinocytes. J Invest Dermatol,2011,131(4):990-993.
[13]Silva J N, Galmiche A, Tome J P, et al. Chain-dependent photocytotoxicity of tricationic porphyrin conjugates and related mechanisms of cell death in proliferating human skin keratinocytes. Biochem Pharmacol,2010,80(9):1373-1385.
[14]von Bultzingslowen I, Jontell M, Hurst P, et al.5-Fluorouracil induces autophagic degeneration in rat oral keratinocytes. Oral Oncol,2001,37(6):537-544.
[15]Lee H M, Shin D M, Yuk J M, et al. Autophagy negatively regulates keratinocyte inflammatory responses via scaffolding protein p62/SQSTM1. J Immunol,2011,186(2): 1248-1258.
[16]Nakatani T, Beitner H. A transmission electron microscopic study of macromelanosomes in sporadic dysplastic naevi. Okajimas Folia Anat Jpn,1992,69(2-3): 109-114.
[17]Hirone T, Eryu Y. Ultrastructure of giant pigment granules in lentigo simplex. Acta Derm Venereol,1978,58(3):223-229.
[1]Harman K E, Albert S, Black M M. Guidelines for the management of pemphigus vulgaris[J]. Br J Dermatol;2003,149(5):926-937.
[2]Hashimoto T. Treatment strategies for pemphigus vulgaris in Japan[J]. Expert Opin Pharmacother,2008,9(9):1519-1530.
[3]Fox L P, Pandya A G. Pulse intravenous cyclophosphamide therapy for dermatologic disorders[J]. Dermatol Clin,2000,18(3):459-473.
[4]Olszewska M, Kolacinska-Strasz Z, Sulej J, et al. Efficacy and safety of cyclophosphamide, azathioprine, and cyclosporine (ciclosporin) as adjuvant drugs in pemphigus vulgaris[J]. Am J Clin Dermatol,2007,8(2):85-92.
[5]Cummins D L, Mimouni D, Anhalt G J, et al. Oral cyclophosphamide for treatment of pemphigus vulgaris and foliaceus[J]. J Am Acad Dermatol,2003,49(2):276-280.
[6]Pasricha J S, Khaitan B K, Raman R S, et al. Dexamethasone-cyclophosphamide pulse therapy for pemphigus[J]. Int J Dermatol,1995,34(12):875-882.
[7]Pandya A G, Sontheimer R D. Treatment of pemphigus vulgaris with pulse intravenous cyclophosphamide[J]. Arch Dermatol,1992,128(12):1626-1630.
[8]Momeni A Z, Iraji F, Aminjavaheri M, et al. The use of oral cyclophosphamide with dexamethasone pulse therapy in the treatment of pemphigus vulgaris[J]. J Dermatolog Treat,2007,18(5):275-278.
[9]Hayag M V, Cohen J A, Kerdel F A. Immunoablative high-dose cyclophosphamide without stem cell rescue in a patient with pemphigus vulgaris[J]. J Am Acad Dermatol,2000,43(6):1065-1069.
[10]Espana A A, Panizo C, Fernandez S, et al. [Prolonged complete clinical remission in patients with severe pemphigus vulgaris after cycles of intravenous cyclophosphamide][J]. Actas Dermosifiliogr,2009,100(2):113-120.
[11]Kanwar A J, De D. Pemphigus in India[J]. Indian J Dermatol Venereol Leprol,2011, 77(4):439-449.
[12]Tirado-Sanchez A, Leon-Dorantes G. Treatment of pemphigus vulgaris. An overview in Mexico[J]. Allergol Immunopathol (Madr),2006,34(1):10-16.
[13]Kanwar A J, Kaur S, Thami G P. Long-term efficacy of dexamethasone-cyclophosphamide pulse therapy in pemphigus [J]. Dermatology,2002, 204(3):228-231.
[14]Martin L K, Werth V P, Villaneuva E V, et al. A systematic review of randomized controlled trials for pemphigus vulgaris and pemphigus foliaceus[J]. J Am Acad Dermatol,2011,64(5):903-908.
[15]Martin L K, Werth V, Villanueva E, et al. Interventions for pemphigus vulgaris and pemphigus foliaceus[J]. Cochrane Database Syst Rev,2009(1):D6263.
[16]Scully C, Challacombe S J. Pemphigus vulgaris:update on etiopathogenesis, oral manifestations, and management[J]. Crit Rev Oral Biol Med,2002,13(5):397-408.
[17]Yeh S W, Sami N, Ahmed R A. Treatment of pemphigus vulgaris:current and emerging options[J]. Am J Clin Dermatol,2005,6(5):327-342.
[18]Kaur S, Kanwar A J. Dexamethasone-cyclophosphamide pulse therapy in pemphigus[J]. Int J Dermatol,1990,29(5):371-374.
[19]Sethy P K, Khandpur S, Sharma V K. Randomized open comparative trial of dexamethasone-cyclophosphamide pulse and daily oral cyclophosphamide versus cyclophosphamide pulse and daily oral prednisolone in pemphigus vulgarisfJ]. Indian J Dermatol Venereol Leprol,2009,75(5):476-482.
[20]邓一文.治疗天疱疮的循证医学研究进展[J].临床口腔医学杂志,2010,26(11)：692-696.
[21]李薇,熊琳,卫茂玲.我国天疱疮治疗性研究的文献评价[J].华西医学,2005,20(4)：624-625.
[22]Mignogna M D.Lo M L, Mignogna R E, et al. Oral pemphigus:long term behaviour and clinical response to treatment with deflazacort in sixteen cases[J]. J Oral Pathol Med,2000,29(4):145-152.
[23]Jain R, Kumar B. Immediate and delayed complications of dexamethasone cyclophosphamide pulse (DCP) therapy[J]. J Dermatol,2003,30(10):713-718.
[24]Mabrouk D, Gurcan H M, Keskin D B, et al. Association between cancer and immunosuppressive therapy--analysis of selected studies in pemphigus and pemphigoid[J]. Ann Pharmacother,2010,44(11):1770-1776.
[25]Zivanovic D, Medenica L, Tanasilovic S, et al. Dexamethasone-cyclophosphamide pulse therapy in pemphigus:a review of 72 cases[J]. Am J Clin Dermatol,2010,11(2): 123-129.
[26]Ramam M. Prolonged antimicrobial and oral cyclophosphamide therapy in pemphigus:need for caution[J]. Indian J Dermatol Venereol Leprol,2009,75(1):85.
[1]Hanifin J M, Cooper K D, Ho V C, et al. Guidelines of care for atopic dermatitis, developed in accordance with the American Academy of Dermatology (AAD)/American Academy of Dermatology Association "Administrative Regulations for Evidence-Based Clinical Practice Guidelines"[J]. J Am Acad Dermatol,2004,50(3):391-404.
[2]Akdis C A, Akdis M, Bieber T, et al. Diagnosis and treatment of atopic dermatitis in children and adults:European Academy of Allergology and Clinical Immunology/American Academy of Allergy, Asthma and Immunology/PRACTALL Consensus Report[J]. Allergy,2006,61(8):969-987.
[3]中华医学会皮肤性病学分会免疫学组.中国特应性皮炎诊断和治疗指南[J].中华皮肤科杂志,2008,41(11)：772-773.
[4]Saeki H, Furue M, Furukawa F, et al. Guidelines for management of atopic dermatitis[J]. J Dermatol,2009,36(10):563-577.
[5]Darsow U, Wollenberg A, Simon D, et al. ETFAD/EADV eczema task force 2009 position paper on diagnosis and treatment of atopic dermatitis[J]. J Eur Acad Dermatol Venereol,2010,24(3):317-328.
[6]Ring J, Alomar A, Bieber T, et al. Guidelines for Treatment of Atopic Eczema (Atopic Dermatitis). For the European Dermatology Forum (EDF), and the European Academy of Dermato-Venereology (EADV), and European Federation of Allergy (EFA), and the European Society of Pediatric Dermatology (ESPD).2011.UD/JR,26.04.2011.
[7]顾恒,陈祥生,陈崑.特应性皮炎诊断标准的评价[J].中华皮肤科杂志,2000,33(4)：222.
[8]Gu H, Chen X S, Chen K, et al. Evaluation of diagnostic criteria for atopic dermatitis: validity of the criteria of Williams et al. in a hospital-based setting[J]. Br J Dermatol,2001,145(3):428-433.
[9]Severity scoring of atopic dermatitis:the SCORAD index. Consensus Report of the European Task Force on Atopic Dermatitis[J]. Dermatology,1993,186(1):23-31.