中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2015年
6期
391-394
,共4页
徐倩倩%陈旭%李莉%徐松%丁兰%张云%鞠梅%李新宇%施辛
徐倩倩%陳旭%李莉%徐鬆%丁蘭%張雲%鞠梅%李新宇%施辛
서천천%진욱%리리%서송%정란%장운%국매%리신우%시신
紫外线%细胞,培养的%角蛋白细胞%信号传导%MAP激酶激酶激酶类
紫外線%細胞,培養的%角蛋白細胞%信號傳導%MAP激酶激酶激酶類
자외선%세포,배양적%각단백세포%신호전도%MAP격매격매격매류
Ultraviolet rays%Cells,cultured%Keratinocytes%Signal transduction%MAP kinase kinase kinases
目的 探讨中波紫外线照射后不同时间点体外培养的HaCaT细胞MAPK信号通路受调控效应.方法 1.5、4.5、7.5、10、20、30、50 mJ/cm2中波紫外线照射HaCaT细胞后8h,对照组细胞除不进行UVB照射外,其他处理同各剂量UVB组,蛋白免疫印迹法测定ERK1/2、JNK和p38蛋白表达及磷酸化水平;50 mJ/cm2中波紫外线照射HaCaT细胞,蛋白免疫印迹法测定照射后2、4、8、12h4个时间点,细胞ERK1/2、JNK和p38蛋白表达及磷酸化水平.使用Quantity One软件计算目的条带吸光度,以相应蛋白条带的吸光度值与GAPDH蛋白内参条带吸光度值的比值表示相对蛋白含量.结果 7.5、10、20、30、50 mJ/cm2 UVB照射HaCaT细胞后8h,ERK1/2、JNK磷酸化水平明显上调(P＜0.05),20、30、50 mJ/cm2 UVB照射HaCaT细胞后p38磷酸化水平上调显著(P＜0.05),且都在50 mJ/cm2照射后效应最为显著(P＜0.05).50 mJ/cm2 UVB照射HaCaT细胞后8h,ERK1/2磷酸化产物水平出现上调,在处理后的12h,与对照组(10.277±0.320)比较,差异有统计学意义(44.844±2.023,P＜ 0.05),而p38和JNK磷酸化产物水平在照射后2h即开始上调(P＜0.05),4、8、12h,p38和JNK与对照相比,UVB照射后虽存在着显著地磷酸化水平差异(P＜0.05),但随时间推移JNK磷酸化产物水平这种上调的趋势逐渐弱化(P＜0.05).结论 在50 mJ/cm2中波紫外线照射后的HaCaT细胞中,ERK1/2、p38、JNK这3种MAPK信号通路产生了活化效应,具有一定的时间效应差异.
目的 探討中波紫外線照射後不同時間點體外培養的HaCaT細胞MAPK信號通路受調控效應.方法 1.5、4.5、7.5、10、20、30、50 mJ/cm2中波紫外線照射HaCaT細胞後8h,對照組細胞除不進行UVB照射外,其他處理同各劑量UVB組,蛋白免疫印跡法測定ERK1/2、JNK和p38蛋白錶達及燐痠化水平;50 mJ/cm2中波紫外線照射HaCaT細胞,蛋白免疫印跡法測定照射後2、4、8、12h4箇時間點,細胞ERK1/2、JNK和p38蛋白錶達及燐痠化水平.使用Quantity One軟件計算目的條帶吸光度,以相應蛋白條帶的吸光度值與GAPDH蛋白內參條帶吸光度值的比值錶示相對蛋白含量.結果 7.5、10、20、30、50 mJ/cm2 UVB照射HaCaT細胞後8h,ERK1/2、JNK燐痠化水平明顯上調(P＜0.05),20、30、50 mJ/cm2 UVB照射HaCaT細胞後p38燐痠化水平上調顯著(P＜0.05),且都在50 mJ/cm2照射後效應最為顯著(P＜0.05).50 mJ/cm2 UVB照射HaCaT細胞後8h,ERK1/2燐痠化產物水平齣現上調,在處理後的12h,與對照組(10.277±0.320)比較,差異有統計學意義(44.844±2.023,P＜ 0.05),而p38和JNK燐痠化產物水平在照射後2h即開始上調(P＜0.05),4、8、12h,p38和JNK與對照相比,UVB照射後雖存在著顯著地燐痠化水平差異(P＜0.05),但隨時間推移JNK燐痠化產物水平這種上調的趨勢逐漸弱化(P＜0.05).結論 在50 mJ/cm2中波紫外線照射後的HaCaT細胞中,ERK1/2、p38、JNK這3種MAPK信號通路產生瞭活化效應,具有一定的時間效應差異.
목적 탐토중파자외선조사후불동시간점체외배양적HaCaT세포MAPK신호통로수조공효응.방법 1.5、4.5、7.5、10、20、30、50 mJ/cm2중파자외선조사HaCaT세포후8h,대조조세포제불진행UVB조사외,기타처리동각제량UVB조,단백면역인적법측정ERK1/2、JNK화p38단백표체급린산화수평;50 mJ/cm2중파자외선조사HaCaT세포,단백면역인적법측정조사후2、4、8、12h4개시간점,세포ERK1/2、JNK화p38단백표체급린산화수평.사용Quantity One연건계산목적조대흡광도,이상응단백조대적흡광도치여GAPDH단백내삼조대흡광도치적비치표시상대단백함량.결과 7.5、10、20、30、50 mJ/cm2 UVB조사HaCaT세포후8h,ERK1/2、JNK린산화수평명현상조(P＜0.05),20、30、50 mJ/cm2 UVB조사HaCaT세포후p38린산화수평상조현저(P＜0.05),차도재50 mJ/cm2조사후효응최위현저(P＜0.05).50 mJ/cm2 UVB조사HaCaT세포후8h,ERK1/2린산화산물수평출현상조,재처리후적12h,여대조조(10.277±0.320)비교,차이유통계학의의(44.844±2.023,P＜ 0.05),이p38화JNK린산화산물수평재조사후2h즉개시상조(P＜0.05),4、8、12h,p38화JNK여대조상비,UVB조사후수존재착현저지린산화수평차이(P＜0.05),단수시간추이JNK린산화산물수평저충상조적추세축점약화(P＜0.05).결론 재50 mJ/cm2중파자외선조사후적HaCaT세포중,ERK1/2、p38、JNK저3충MAPK신호통로산생료활화효응,구유일정적시간효응차이.
Objective To explore the regulatory effects of ultraviolet B (UVB) radiation on the mitogenactivated protein kinase (MAPK) signaling pathway in cultured HaCaT cells in vitro at different post-radiation time points.Methods Some cultured HaCaT cells were classified into several groups to be exposed to UVB of 1.5,4.5,7.5,10,20,30 and 50 mJ/cm2 for 1,3,5,7,15,20 and 34 seconds respectively,or UVB of 50 mJ/cm2 for 34 seconds,or remain untreated (control group).Western blot was performed to determine the total and phosphorylation levels of ERK1/2,JNK and p38 in HaCaT cells at 8 hours after exposure to 1.5,4.5,7.5,10,20,30 and 50 mJ/cm2 UVB,as well as at 2,4,8 and 12 hours after exposure to 50 mJ/cm2 UVB.Quantity One software was used to measure the absorbance value of protein bands,and protein levels were expressed as the absorbance ratio of target bands to glyceraldehyde-3-phosphate dehydrogenase (GAPDH).Statistical analysis was carried out by using one-way analysis of variance,least significant difference (LSD)-t test and multivariate analysis of variance.Results Compared with the control group,there was a significant increase at 8 hours in the phosphorylation levels of ERK 1/2 and JNK in HaCaT cells after exposure to 7.5,10,20,30 and 50 mJ/cm2 UVB (all P ＜ 0.05),and in the phosphorylation level of p38 after exposure to 20,30 and 50 mJ/cm2 UVB (all P ＜ 0.05),with the strongest increase in phosphorylation levels of ERK 1/2,JNK and p38 all observed in HaCaT cells exposed to 50 mJ/cm2 UVB (all P ＜ 0.05).After exposure to 50 mJ/cm2 UVB,the phosphorylation level of ERK1/2 began to increase in HaCaT cells at 8 hours,and significantly increased at 12 hours (44.844 ± 2.023 vs.10.277 ± 0.320,P ＜ 0.05) compared with the control group;however,the phosphorylation levels of p38 and JNK increased in HaCaT cells as early as 2 hours after exposure (both P ＜ 0.05),and remained significantly higher compared with the control group at 4,8 and 12 hours (all P ＜ 0.05),while the increasing trend in phosphorylated JNK level weakened over time (P ＜ 0.05).Conclusions The ERK1/2,JNK and p38 MAPK signaling pathways were activated in HaCaT cells after exposure to 50 mJ/cm2 UVB,and the degree of their activation varied over time.
