中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2013年
3期
176-180
,共5页
王莹%陈旭%鞠梅%姜鹏爽%郭新云%夏立君%蒋靖%陈崑%顾恒
王瑩%陳旭%鞠梅%薑鵬爽%郭新雲%夏立君%蔣靖%陳崑%顧恆
왕형%진욱%국매%강붕상%곽신운%하립군%장정%진곤%고항
紫外线%角蛋白细胞%bcl-2相关X蛋白质%半胱氨酸天冬氨酸蛋白酶-3%Beclin-1
紫外線%角蛋白細胞%bcl-2相關X蛋白質%半胱氨痠天鼕氨痠蛋白酶-3%Beclin-1
자외선%각단백세포%bcl-2상관X단백질%반광안산천동안산단백매-3%Beclin-1
Ultraviolet rays%Keratinocyte%bcl-2-Associated X protein%Caspase 3%Beclin-1
目的 观察慢性紫外线损伤对小鼠皮肤角质形成细胞凋亡和自噬交互调控元件Bax、Bcl-2、Beclin-1及半胱氨酸天冬氨酸蛋白酶3(caspase3)的调控效应.方法 健康昆明小鼠30只,雌雄各半,鼠龄6～8周,按性别随机分成慢性紫外线损伤组(20只)和对照组(10只),每组雌雄各半,根据性别将小鼠分笼饲养.采用模拟日光紫外线(UVA+ UVB)光源对实验小鼠进行照射,从最小红斑量(UVB 0.07 J/cm2,UVA 0.7 J/cm2)开始,每周增加0.5个最小红斑量,每日1次,每周照射5d,共9周,UVB总剂量达9.45 J/cm2,UVA总剂量达94.5 J/cm2.通过组织学观察、特殊化学染色和表皮厚度测量确定皮肤损伤,应用免疫组化法分别对照射前(W0)和实验9周末(W9)Bax、Bcl-2、Beclin-1和caspase3的表达进行检测,表达强度以免疫反应强度分布指数(IRIDI)表示,并与对照组进行比对分析.分别采用配对t检验和Wilcoxon符号秩和检验对各参数进行比较.结果 慢性紫外线照射后,小鼠表皮厚度明显增加.肉眼观察、组织学观察和胶原纤维、弹性纤维特殊染色均显示损伤组小鼠皮肤临床表现和组织学改变均与慢性紫外线光损伤的人类皮肤较为吻合.在损伤组,照光前Beclin-1、caspase3、Bax、Bcl-2的表达计分均值分别为0.30、0.25、0.35、0.25,照光后分别为2.70、3.30、3.35、0.25.Beclin-1、caspase3、Bax蛋白表达显著升高,且差异有统计学意义(Z值分别为4.034、4.001、3.970,均P＜ 0.05),Bcl-2蛋白表达变化不明显(Z=0,P＞ 0.05).对照组小鼠Beclin-1、caspase3、Bax、Bcl-2的均值在W0时和W9时表达差异均无统计学意义(Z值分别为0、0.577、0、0.577,均P＞0.05).结论 慢性紫外线损伤对小鼠皮肤角质形成细胞中凋亡和自噬的交互调控元件Beclin-1和Bax具有上调表达的作用,而对Bcl-2表达调控效应不显著.这一系列的调控效应可能参与了凋亡和自噬在慢性紫外线皮肤损伤中的交互调控.
目的 觀察慢性紫外線損傷對小鼠皮膚角質形成細胞凋亡和自噬交互調控元件Bax、Bcl-2、Beclin-1及半胱氨痠天鼕氨痠蛋白酶3(caspase3)的調控效應.方法 健康昆明小鼠30隻,雌雄各半,鼠齡6～8週,按性彆隨機分成慢性紫外線損傷組(20隻)和對照組(10隻),每組雌雄各半,根據性彆將小鼠分籠飼養.採用模擬日光紫外線(UVA+ UVB)光源對實驗小鼠進行照射,從最小紅斑量(UVB 0.07 J/cm2,UVA 0.7 J/cm2)開始,每週增加0.5箇最小紅斑量,每日1次,每週照射5d,共9週,UVB總劑量達9.45 J/cm2,UVA總劑量達94.5 J/cm2.通過組織學觀察、特殊化學染色和錶皮厚度測量確定皮膚損傷,應用免疫組化法分彆對照射前(W0)和實驗9週末(W9)Bax、Bcl-2、Beclin-1和caspase3的錶達進行檢測,錶達彊度以免疫反應彊度分佈指數(IRIDI)錶示,併與對照組進行比對分析.分彆採用配對t檢驗和Wilcoxon符號秩和檢驗對各參數進行比較.結果 慢性紫外線照射後,小鼠錶皮厚度明顯增加.肉眼觀察、組織學觀察和膠原纖維、彈性纖維特殊染色均顯示損傷組小鼠皮膚臨床錶現和組織學改變均與慢性紫外線光損傷的人類皮膚較為吻閤.在損傷組,照光前Beclin-1、caspase3、Bax、Bcl-2的錶達計分均值分彆為0.30、0.25、0.35、0.25,照光後分彆為2.70、3.30、3.35、0.25.Beclin-1、caspase3、Bax蛋白錶達顯著升高,且差異有統計學意義(Z值分彆為4.034、4.001、3.970,均P＜ 0.05),Bcl-2蛋白錶達變化不明顯(Z=0,P＞ 0.05).對照組小鼠Beclin-1、caspase3、Bax、Bcl-2的均值在W0時和W9時錶達差異均無統計學意義(Z值分彆為0、0.577、0、0.577,均P＞0.05).結論 慢性紫外線損傷對小鼠皮膚角質形成細胞中凋亡和自噬的交互調控元件Beclin-1和Bax具有上調錶達的作用,而對Bcl-2錶達調控效應不顯著.這一繫列的調控效應可能參與瞭凋亡和自噬在慢性紫外線皮膚損傷中的交互調控.
목적 관찰만성자외선손상대소서피부각질형성세포조망화자서교호조공원건Bax、Bcl-2、Beclin-1급반광안산천동안산단백매3(caspase3)적조공효응.방법 건강곤명소서30지,자웅각반,서령6～8주,안성별수궤분성만성자외선손상조(20지)화대조조(10지),매조자웅각반,근거성별장소서분롱사양.채용모의일광자외선(UVA+ UVB)광원대실험소서진행조사,종최소홍반량(UVB 0.07 J/cm2,UVA 0.7 J/cm2)개시,매주증가0.5개최소홍반량,매일1차,매주조사5d,공9주,UVB총제량체9.45 J/cm2,UVA총제량체94.5 J/cm2.통과조직학관찰、특수화학염색화표피후도측량학정피부손상,응용면역조화법분별대조사전(W0)화실험9주말(W9)Bax、Bcl-2、Beclin-1화caspase3적표체진행검측,표체강도이면역반응강도분포지수(IRIDI)표시,병여대조조진행비대분석.분별채용배대t검험화Wilcoxon부호질화검험대각삼수진행비교.결과 만성자외선조사후,소서표피후도명현증가.육안관찰、조직학관찰화효원섬유、탄성섬유특수염색균현시손상조소서피부림상표현화조직학개변균여만성자외선광손상적인류피부교위문합.재손상조,조광전Beclin-1、caspase3、Bax、Bcl-2적표체계분균치분별위0.30、0.25、0.35、0.25,조광후분별위2.70、3.30、3.35、0.25.Beclin-1、caspase3、Bax단백표체현저승고,차차이유통계학의의(Z치분별위4.034、4.001、3.970,균P＜ 0.05),Bcl-2단백표체변화불명현(Z=0,P＞ 0.05).대조조소서Beclin-1、caspase3、Bax、Bcl-2적균치재W0시화W9시표체차이균무통계학의의(Z치분별위0、0.577、0、0.577,균P＞0.05).결론 만성자외선손상대소서피부각질형성세포중조망화자서적교호조공원건Beclin-1화Bax구유상조표체적작용,이대Bcl-2표체조공효응불현저.저일계렬적조공효응가능삼여료조망화자서재만성자외선피부손상중적교호조공.
Objective To study the regulatory effect of chronic ultraviolet (UV) radiation on the expressions of three regulatory elements involved in the cross-talk between apoptosis and autophagy (including Bax,Bcl-2 and Beclin-1),as well as cysteine-containing aspartate-specific protease 3 (caspase 3) in keratinocytes of mouse skin.Methods Thirty healthy Kunming mice at 6-8 weeks of age were included in this study,and randomly divided into two groups with the ratio of female to male mice being 1 ∶ 1 in each group:chronic UV damage group (n =20) receiving sunlight simulator UV radiation,and control group (n =10) receiving no radiation.The irradiation was performed on the back of mice once a day for 5 consecutive days per week,and lasted for 9 weeks.The dose of irradiation began at one minimal erythema dose (UVB:0.07 J/cm2,UVA:0.7 J/cm2),and increased by 0.5 minimal erythema dose per week,with the total dose of UVB and UVA being 9.45 J/cm2 and 94.5 J/cm2 respectively.Skin samples were resected from the back of these mice before and at 9 weeks after the beginning of the radiation.Skin damage was evaluated by histological observation,specific chemical staining and epidermal thickness measurement.Immunohistochemistry was conducted to detect the expressions of Bax,Bcl-2,caspase 3 and Beclin-1 in epidermal keratinocytes.Paired t test and Wilcoxon signed rank test were carried out for statistical analysis.Results After UV irradiation,the mice showed an obvious increase in epidermal thickness.Histological examination and specific staining for collagen and elastic fibers both revealed a histological change characteristic of chronic UV damage in the mice receiving UV radiation.In the chronic UV damage group,the average immunoreactivity intensity distribution index (IRIDI) was statistically increased for Beclin-1,caspase 3 and Bax in the skin sample (2.70 vs.0.30,3.30 vs.0.25,3.35 vs.0.35,Z =4.034,4.001,3.970,respectively,all P ＜ 0.05),but remained unchanged for Bcl-2 (0.25 vs.0.25,Z =0,P ＞0.05) at 9 weeks after the radiation compared with that before radiation.No significant changes were observed in the control group for the expression intensity of Beclin-1,caspase 3,Bax or Bcl-2 during the 9 weeks (Z =0,0.577,0,0.577,respectively,all P ＞ 0.05).Conclusions Chronic UV irridiation shows no obvious effect on the expression of Bcl-2,but can up-regulate the expression of Beclin-1 and Bax,both of which may be involved in the cross-talk between apoptosis and autophagy in chronic UV radiation-induced skin damage.
