知识与财富的链接
【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17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（pcDNA31（+）/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＜005),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＜005）。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＜001）。pcDNA31（+）/K17对KC、HaCaT细胞凋亡率的抑制作用被EGCG显著逆转（P＜001）。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。
【Abstract】 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 (pcDNA31（+）/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＜005) 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＜005). EGCG (50 mg/L ) increased the apoptosis rate of KC, HaCaT and HPV11.HaCaT (P＜001). EGCG attenuated the role of pcDNA3. 1（+）/K17 on cell apoptosis of KC and HaCaT (P＜001).Conclusion EGCG promotes keratinocyte apoptosis by inhibiting K17, which provides a basis for the basic research and clinical treatment of psoriasis and condyloma acuminatum.
