莪术醇可能通过Notch信号通路影响角质形成细胞的增殖、迁移、凋亡  

Curcumol may regulate the mechanism of Notch signaling pathway on the proliferation,migration and apoptosis of keratinocytes

作　　者：梁博[1] 李永辉[1] 孙继玮 许进[1] LIANG Bo;LI Yonghui;SUN Jiwei;XU Jin(Department of Dermatology,Baoding Second Hospital,Baoding 071051,China)

机构地区：[1]保定市第二医院皮肤科,河北保定071051

出　　处：《东南大学学报（医学版）》2023年第3期339-347,共9页Journal of Southeast University(Medical Science Edition)

基　　金：河北省保定市社会发展类项目(18ZF037)。

摘　　要：目的:探讨莪术醇对角质形成细胞增殖、迁移、凋亡的作用及其相关的分子机制。方法:体外分离培养银屑病角质形成细胞,使用浓度0、10、40、80 mg·L^(-1)莪术醇处理细胞,CCK8实验检测细胞增殖;流式细胞术检测细胞凋亡;蛋白质印迹法检测细胞周期蛋白D1(Cyclin D1)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Notch 1、Hes 1蛋白表达水平;Transwell小室、伤口愈合实验检测细胞迁移。以0 mg·L^(-1)莪术醇作为对照,将80 mg·L^(-1)莪术醇、Notch信号通路抑制剂DAPT加入80 mg·L^(-1)莪术醇处理的细胞内,通过上述方法检测加入Notch通路抑制剂对银屑病角质形成细胞增殖、迁移和凋亡的影响。多组间比较通过单因素方差分析,组内间比较使用LSD-t检验。结果:莪术醇呈剂量效应降低银屑病角质形成细胞活性、迁移率、迁移细胞数和Cyclin D1、Bcl-2蛋白表达,增加凋亡率和Bax、Cleaved-Caspase3、Cleaved-Caspase9、Notch 1、Hes 1蛋白表达(P<0.05)。与莪术醇组比较,莪术醇+DAPT组细胞活性、迁移率、迁移细胞数和Cyclin D1、Bcl-2蛋白表达增加,凋亡率和Bax、Cleaved-Caspase3、Cleaved-Caspase9蛋白表达水平降低,差异有统计学意义(P<0.05)。结论:莪术醇可能通过激活Notch信号通路抑制银屑病角质形成细胞的增殖、迁移,并诱导细胞凋亡。Objective:To investigate the effects of curcumol on the proliferation,migration and apoptosis of keratinocytes and its related molecular mechanisms.Methods:Psoriasiskeratinocytes were isolated and cultured in vitro,treated with curcumol concentrations of 0,10,40,and 80 mg·L^(-1),and cell counting kit(CCK8)experiment was used to detect cell proliferation;flow cytometry experiment was used to detect cell apoptosis;Western blot experiments were used to detect cyclin D1(Cyclin D1),B cell lymphoma/leukemia-2(Bcl-2),Bcl-2-related X protein(Bax),Notch 1,and Hes 1 protein expression levels;Transwell chambers and wound healing experiments were used to detect cell migration.With 0 mg·L^(-1)curcumol as control,80 mg·L^(-1)curcumol and Notch signaling pathway inhibitor DAPT were added into the cells treated with 80 mg·L^(-1)curcumol,and the effects of adding Notch pathway inhibitors on the proliferation,migration and apoptosis of psoriasis keratinocytes were detected by the above method.Comparisons among multiple groups were performed by one-way ANOVA,and LSD-t was used for within-group comparison.Results:Curcumol decreased keratinocyte activity,migration rate,number of migrating cells,Cyclin D1,Bcl-2 protein expression,and increased apoptosis rate and expression of Bax,Cleaved-Caspase3,Cleaved-Caspase9,Notch 1,Hes 1 protein in psoriasis with a dose-dependent manner(P<0.05).Compared with the curcumol group,the cell activity,migration rate,number of migrated cells,and expression of Cyclin D1 and Bcl-2 proteins in the curcumol+DAPT group increased,while the apoptosis rate and Bax,Cleaved-Caspase3,Cleaved-Caspase9 protein expression levels decreased,with a statistically significant difference(P<0.05).Conclusion:Curcumol may inhibit the proliferation and migration of psoriatic keratinocytes and induce cell apoptosis by activating Notch signaling pathway.

关 键 词：莪术醇 NOTCH信号通路 角质形成细胞 增殖 迁移 凋亡 

分 类 号：R-33[医药卫生]