表没食子儿茶素没食子酸酯通过抑制角蛋白17诱导角质形成细胞凋亡  

Epigallocatechin gallate induces the apoptosis of malpighian cells via inhibiting keratin 17

作　　者：朱自荣 王飞苗[2] 赵生魁 潘萍 雷德军 金小雅 ZHU Zirong;WANG Feimiao;ZHAI Shengkui;PAN Ping;LEI Dejun;JIN Xiaoya(The Eighth hospital of Dermatology and Venereology of Xi’an,Xi'an710004,China;Reproductive Medicine Center, General Hospital of Ningxia Medical University,Yinchuan 750000,China;Department of Skin Disease and Venereal Disease, School of Medicine,Xi'an Jiaotong University,Xi'an 710000,China;Dermatological Department, Shanxi Province Friendship Hospital,Xi'an 710068,China)

机构地区：[1]西安市第八医院皮肤病与性病科,陕西西安710004 [2]宁夏医科大学总医院生殖医学中心,宁夏银川750000 [3]西安交通大学医学院皮肤病与性病学科,陕西西安710000 [4]陕西省友谊医院皮肤科,陕西西安710068

出　　处：《西部医学》2019年第6期857-862,共6页Medical Journal of West China

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)对角蛋白17(K17)介导的人角质形成细胞的凋亡的影响.方法不同浓度EGCG处理人乳头瘤病毒(HPV)11型基因组的HaCaT细胞(HPV11.HaCaT)12 h,qRT-PCR与Western blot检测角蛋白1 7表达;流式细胞术Annexin V-FITC和碘化丙锭(PI)双染检测50 mg/L EGCG处理下KC、HaCaT及HPV11.HaCaT细胞凋亡率.免疫组化法与Western blot检测银屑病和尖锐湿疣患者病理切片中K17以及Caspase3的表达,构建K17过表达(pcDNA3.1(+)/K17)载体并转染KC与HaCaT细胞48 h继续用EGCG(50 mg/L)处理12 h;检测K17的水平及细胞凋亡.结果 EGCG可在12 h抑制KC、HaCaT和HPV11.HaCaT细胞中K17表达(P<0.05),呈浓度依赖趋势;银屑病和尖锐湿疣病理切片中K17表达范围增多但Caspase3的染色范围减少,K17表达上调,但Cleaved-Caspase3显著下调(P<0.05).使用高浓度50 mg/L的EGCG作用下,KC、HaCaT和HPV11.HaCaT细胞凋亡率均上调(P<0.01).pcDNA3.1 (+)/K17对KC、HaCaT细胞凋亡率的抑制作用被EGCG显著逆转(P<0.01).结论 EGCG通过抑制K17促进角质形成细胞凋亡,为银屑病和尖锐湿疣基础研究与临床治疗提供前期研究基础.Objective To investigate the effects of EGCG on keratin 17(K17)-mediated human keratinocyte apoptosis. Methods Different concentrations of EGCG were treated with HPV11.HaCaT for 12 h. The expression of K17 was tested by qRT-PCR and Western blot. Flow cytometry(Annexin V-FITC and PI double staining) was used to study the apoptosis rates of KC, HaCaT and HPV11.HaCaT cells. The expression levels of K17 and Caspase 3 in pathological sections of patients with psoriasis and condyloma acuminatum were tested by immunohistochemistry and Western blot method. Construction of K17 overexpression vectors(pcDNA3.1(+)/K17), and the vectors were transfected in KC and HaCaT for 48 h, and further EGCG(50 mg/L)was treated the cells for 12 h. The level of K17 and apoptosis were detected. Results The expression of K17 was significantly decreased in KC, HaCaT and HPV11.HaCaT(P<0.05) by EGCG treatment at 12 h with a concentration dependent trend. The positive rate of K17 in psoriasis and condyloma acuminatum was increased, but the positive rate of Caspase3 was decreased. However, the expression of Cleaved-Caspase 3 was significantly downregulated(P<0.05). EGCG(50 mg/L) increased the apoptosis rate of KC, HaCaT and HPV11.HaCaT(P<0.01). EGCG attenuated the role of pcDNA3. 1(+)/K17 on cell apoptosis of KC and HaCaT(P<0.01).Conclusion EGCG promotes keratinocyte apoptosis by inhibiting K17, which provides a basis for the basic research and clinical treatment of psoriasis and condyloma acuminatum.

关 键 词：表没食子儿茶素没食子酸酯 角蛋白17 角质形成细胞 凋亡 

分 类 号：R961[医药卫生—药理学]