中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
6期
400-403
,共4页
张敏%苗叶%薛柯%李承新
張敏%苗葉%薛柯%李承新
장민%묘협%설가%리승신
HaCaT细胞%瘦素%角蛋白17
HaCaT細胞%瘦素%角蛋白17
HaCaT세포%수소%각단백17
HaCaT cells%Leptin%Keratin-17
目的 探讨瘦素对HaCaT细胞角蛋白17(K17)表达的影响.方法 体外培养HaCaT细胞,给予100 ng/ml的瘦素作用24 h,应用实时PCR检测K17 mRNA表达水平、Western印迹及免疫荧光染色法检测K17蛋白表达水平变化.结果与阴性对照组(1.000 0±0.000 0)相比较,瘦素组(3.086 7±0.186 1)K17mRNA表达显著升高,差异有统计学意义(P＜0.01).Western印迹结果表明,瘦素组K17蛋白较阴性对照组显著上调,细胞免疫荧光染色结果与RT-PCR、Western印迹结果相符.与单纯使用瘦素组(2.242 7±0.188 7)相比较,STAT3抑制剂组和Erk 1/2抑制剂组K17 mRNA分别为0.674 1±0.060 0、0.855 0±0.390 3,Western印迹和细胞免疫荧光染色显示,两个抑制剂组的K17蛋白较瘦素组均显著下调,差异均有统计学意义(P＜0.01).结论瘦素可以诱导HaCaT细胞表达K17,其机制可能与激活STAT3、Erk 1/2信号转导途径有关.
目的 探討瘦素對HaCaT細胞角蛋白17(K17)錶達的影響.方法 體外培養HaCaT細胞,給予100 ng/ml的瘦素作用24 h,應用實時PCR檢測K17 mRNA錶達水平、Western印跡及免疫熒光染色法檢測K17蛋白錶達水平變化.結果與陰性對照組(1.000 0±0.000 0)相比較,瘦素組(3.086 7±0.186 1)K17mRNA錶達顯著升高,差異有統計學意義(P＜0.01).Western印跡結果錶明,瘦素組K17蛋白較陰性對照組顯著上調,細胞免疫熒光染色結果與RT-PCR、Western印跡結果相符.與單純使用瘦素組(2.242 7±0.188 7)相比較,STAT3抑製劑組和Erk 1/2抑製劑組K17 mRNA分彆為0.674 1±0.060 0、0.855 0±0.390 3,Western印跡和細胞免疫熒光染色顯示,兩箇抑製劑組的K17蛋白較瘦素組均顯著下調,差異均有統計學意義(P＜0.01).結論瘦素可以誘導HaCaT細胞錶達K17,其機製可能與激活STAT3、Erk 1/2信號轉導途徑有關.
목적 탐토수소대HaCaT세포각단백17(K17)표체적영향.방법 체외배양HaCaT세포,급여100 ng/ml적수소작용24 h,응용실시PCR검측K17 mRNA표체수평、Western인적급면역형광염색법검측K17단백표체수평변화.결과여음성대조조(1.000 0±0.000 0)상비교,수소조(3.086 7±0.186 1)K17mRNA표체현저승고,차이유통계학의의(P＜0.01).Western인적결과표명,수소조K17단백교음성대조조현저상조,세포면역형광염색결과여RT-PCR、Western인적결과상부.여단순사용수소조(2.242 7±0.188 7)상비교,STAT3억제제조화Erk 1/2억제제조K17 mRNA분별위0.674 1±0.060 0、0.855 0±0.390 3,Western인적화세포면역형광염색현시,량개억제제조적K17단백교수소조균현저하조,차이균유통계학의의(P＜0.01).결론수소가이유도HaCaT세포표체K17,기궤제가능여격활STAT3、Erk 1/2신호전도도경유관.
Objective To evaluate the effect of leptin on K17 expression in HaCaT human keratinocytes.Methods Some cultured HaCaT cells were treated with leptin (100 ng/ml) or remained untreated for 24 hours followed by the quantification of K17 mRNA expression by real-time PCR and detection of K17 protein expression by Western blot and immunofluorescence staining.To investigate the action mechanism of leptin,some cultured HaCaT cells were divided into several groups to be treated with leptin (100 ng/ml) alone,Piceatannol (an inhibitor of the STAT3 pathway) + leptin (100 ng/ml),PD-98059 (an inhibitor of the Erk1/2 pathway) + leptin (100 ng/ml),respectively for 24 hours,with the cells receiving no treatment as the negative control.Subsequently,the mRNA and protein expressions of K17 were measured by the above methods.Statistical analysis was done by the two-sample ttest.Results The mRNA expression of K17 was significantly higher in HaCaT cells treated with leptin alone than in those remaining untreated (3.086 7 ± 0.186 1 vs.1.000 0 ± 0.000 0,P ＜ 0.01),but significantly downregulated in HaCaT cells treated with Piceatannol + leptin and those with PD-98059 + leptin compared with those treated leptin alone (0.674 1 ± 0.060 0 and 0.855 0 ± 0.390 3 vs.2.242 7 ± 0.188 7,both P ＜ 0.01).The results of Western blot and immunofluorescence staining were in agreement with those of real-time PCR.Conclusions Leptin can induce K17 expression in HaCaT cells,likely by activating the STAT3 and Erk1/2 signaling pathways.
