羊毛固醇合成酶抑制剂对KCs细胞分化和凋亡的影响  

Effects of lanosterol synthase inhibitor on differentiation and apoptosis of KCs

作　　者：刘莉 顾亚男 李名聪 黄依璇 张胜权 Liu Li;Gu Yanan;Li Mingcong;Huang Yixuan;Zhang Shengquan(Center for Scientific Research of Anhui Medical University,Dept of Biochemistry and Molecular Biology, College of Basic Medicine,The First College of Clinical Medicine,Hefei 230032)

机构地区：[1]安徽医科大学科研实验中心,基础医学院生物化学与分子生物学教研室,第一临床医学院,合肥230032

出　　处：《安徽医科大学学报》2022年第2期173-177,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271748);安徽省高校自然科学研究项目(编号:KJ2020A0197)。

摘　　要：目的探究单独应用羊毛固醇合成酶抑制剂(RO)或联合胆固醇(CH)刺激皮肤角质形成细胞(KCs)后对其分化及凋亡的影响。方法RO单独或联合CH于KCs共培养不同时间后,加入Ca^(2+)(1.8 mmol/L)处理1d,Western blot法分析KCs的分化相关蛋白角皮蛋白(INV)和兜甲蛋白的表达;RO单独或联合CH与KCs共培养不同时间后,流式细胞术检测KCs细胞凋亡变化和Western blot法验证凋亡蛋白Bax和Bcl-2表达。结果RO下调KCs,Ca^(2+)诱导的分化标志蛋白INV的表达,对Loricrin表达抑制作用较弱,而CH没有表现出对RO的拮抗效应;RO诱导KCs细胞凋亡并呈时间依赖性,CH能够拮抗RO对KCs的诱导凋亡作用;RO单独应用时细胞凋亡相关蛋白Bcl-2和Bax的表达均受到抑制,联合CH应用能部分拮抗RO对Bcl-2表达抑制作用,对Bax的表达抑制无显著影响;RO能时间依赖性降低Bcl-2/Bax的比值,CH能部分减弱RO对Bcl-2/Bax比值的影响。结论RO可能通过下调Loricrin、Bcl-2的表达,进而抑制KCs分化和诱导KCs细胞凋亡。Objective To investigate the effects of RO or incubation with cholesterol(CH)on the differentiation and apoptosis of KCs.Methods Ca^(2+)(1.8 mmol/L)was added to KCs for 1 day after co-culture of RO alone or combined with CH in KCs for different time.The expression of Involucrin(INV)and Loricrin in KCs was analyzed by Western blot.After co-culture of RO alone or combination with CH in KCs for different time,the apoptotic changes of KCs cells were detected by flow cytometry and the expression of apoptotic proteins Bax and Bcl-2 were verified by Western blot.Results RO down-regulated the expression of Ca2+induced differentiation marker protein INV,but weakly inhibited the expression of Loricrin,while CH showed no antagonistic effect on RO.RO induced apoptosis of KCs cells in a time dependent manner.CH could antagonize the apoptotic effect of RO on KCs;the expression of Bcl-2 and Bax was inhibited when RO was applied alone,CH could partially antagonize the inhibition effect of RO on Bcl-2 expression,but had no significant effect on Bax;however,RO reduced the ratio of Bcl-2/Bax in a time-dependent manner,and CH partially weakened the effect of RO on the ratio of Bcl-2/Bax.Conclusion RO may inhibit KCs differentiation and induce KCs cell apoptosis by down-regulating the expression of Loricrin and Bcl-2.

关 键 词：毛固醇合成酶抑制剂 胆固醇 角质形成细胞 细胞分化 细胞凋亡 

分 类 号：R3[医药卫生—基础医学]